Research ArticleExperimental Studies

Effect of Liposomal Curcumin on Acetaminophen Hepatotoxicity by Down-regulation of Oxidative Stress and Matrix Metalloproteinases

GABRIELA DOGARU, ADRIANA ELENA BULBOACA, DAN GHEBAN, PAUL MIHAI BOARESCU, VASILE RUS, DANA FESTILA, ADELA-VIVIANA SITAR-TAUT and IOANA STANESCU
In Vivo March 2020, 34 (2) 569-582; DOI: https://doi.org/10.21873/invivo.11809

Abstract

Background/Aim: The hepatoprotective role of various molecules in drug-induced hepatotoxicity arouses great interest. We investigated the effect of liposomal curcumin (LCC) on experimental acetaminophen (APAP)-induced hepatotoxicity. Materials and Methods: Rats were randomly allocated into 5 groups, and the effect of two LCC concentrations was studied: group 1 – 1 ml intraperitoneal (i.p.) saline, group 2 – APAP pretreatment, group 3 – APAP+silymarin (extract of the silybum marianum with anti-inflammatory, anti-oxidant, and anti-fibrotic properties), group 4 – APAP+LCC1, group 5 – APAP+LCC2. The biomarkers of oxidative stress (nitric oxide and malondialdehyde) and antioxidant status of plasma (thiols and catalase), TNF-α, MMP-2 and MMP-9 serum levels were evaluated. Results: An improvement in oxidative stress, antioxidant status, and TNF-α, MMP-2 and MMP-9 levels was obtained in groups pretreated with LCC compared to silymarin treatment, in a dose-dependent manner. Histopathological examination reinforced the results. Conclusion: Liposomal curcumin improves the oxidative stress/antioxidant balance and alleviates inflammation in experimental APAP-induced hepatotoxicity.

Acetaminophen, also known as paracetamol or N-acetyl-p-aminophenol (APAP), has anti-inflammatory, analgesic and antipyretic properties when used in its normal therapeutic doses (1, 2). It is widely available and can be used by all age groups. The tendency to use it in high doses for more rapid effects is the main concern, due to increased risk for acute liver failure that can be very severe, requiring liver transplantation (1). The liver is a target for the biotransformation of many drugs; therefore, liver tissue lesions are commonly associated with various medications (3). Acetaminophen-related liver toxicity is reported in many studies and is mainly associated with drug overdose, genetic factors, concurrent medication, concomitant alcohol consumption or nutritional status (4). Hepatic tissue lesions can be very severe, leading to acute liver failure (ALF) (4). ALF is related to cell death that has been documented to occur mainly by necrosis, but the apoptotic process can also have an important contribution (5). Inhibition of caspase activity (involved in the apoptotic process) proved to promote liver tissue regeneration and recovery (6).

Mitochondrial dysfunction is one of the mechanisms involved in hepatic toxicity of APAP, consequently inducing hepatic cell death (7). Cell response to mitochondrial dysfunction consists of various deleterious consequences, including excessive production of reactive oxygen species (ROS) (8). Mitochondrial dysfunction has been proven to be related to a decrease in ATP production, an increase in ROS, and a decrease in the production of glutathione (GLUT) as a scavenger molecule for ROS (7). APAP hepatotoxicity is also related to increased production of reactive metabolite N-acetyl-p-benzoquinone (NAPQI) as a consequence of the metabolic activity of the cytochrome P450 system (9). The metabolite depletes GLUT from liver tissue, decreasing the cell's antioxidant defense system (9). GLUT production is important for the hepatic metabolism of APAP, acting as a scavenger for NAPQI protein (8). Changes in cellular enzymatic activity (involved in drug detoxification) can also cause cellular damage to liver tissue (10). Oxidative stress and inflammatory reactions were also incriminated in overdoses of APAP inducing renal toxicity (11). Depletion of the GLUT pool is another important mechanism that contributes to APAP hepatotoxicity (12). Hepatic injury is associated with increased synthesis of matrix metalloproteinases (MMPs), which are a group of proteolytic enzymes that can promote extracellular matrix (ECM) degradation, being a good indicator of liver damage in liver failure (13). MMPs can also act as regulators of inflammation and immunity by influencing cytokine and chemokine production (14). The normal liver expresses several MMPs, including MMP-2 and MMP-9, which have an important role in liver vascular homeostasis (15). Increased MMP-9 activity is associated with extensive leukocyte recruitment in severe liver injury (16). During their activation at the injury site, leukocytes can express MMP-9 (responsible for ECM degradation and for increased vascular permeability due to cleavage of endothelial junctional proteins) (17, 18).

Thus, the major issues in APAP-induced hepatotoxicity are to identify potential biomarkers to predict the severity of liver damage and to develop new therapies able to limit the extension of injury and promote liver regeneration. Natural products are attracting the interest of many researchers to study their effects on various disorders. Among the nutraceutical compounds commonly appreciated for their antioxidant and anti-inflammatory properties is curcumin, which can improve cell viability in various disorders by reducing cell apoptosis and necrosis (19). The antioxidant activity of curcumin is based on its scavenging properties for ROS (20, 21). Its anti-inflammatory effect was reported to be related to the down-regulation of nuclear factor-kB (NF-kB), cyclooxygenase 2 (COX2) and pro-inflammatory cytokines such as interleukin-1 and interleukin-6 (22). We previously demonstrated the beneficial effect of oral curcumin administration on hepatic function in fructose-induced metabolic syndrome and in myocardial ischemia (23-25). Despite the beneficial effect of curcumin, the low bioavailability of this natural compound presents interest for new formulations. Novel nanoformulations of curcumin are emerging and can enhance its systemic bioavailability and tissue distribution (26). Among curcumin nanoformulations, liposomal curcumin offers a better water solubility of curcumin, leading to 8- to 20-fold increased systemic exposure compared to the standard curcumin suspension formulation (27). Liposomal curcumin is a phospholipid bilayer vesicle that can carry curcumin; it is easy to prepare and safe to use (28-30).

View this table:
Table I.

The levels of the investigated markers of liver damage (ALT and AST) and inflammation (TNF-α) in each treatment group, expressed as mean and standard deviation.

Silymarin (SIL) is an active component of Silybum marianum extracts with anti-inflammatory, anti-oxidant, and anti-fibrotic properties. It contains a family of flavonolignans and a flavonoid (taxifolin), among which silybin accounts for 50% to 70% and is identified as the major biologically active component (31).

The aim of this study was to observe the beneficial effect of liposomal curcumin administration in two concentrations and to compare it to the effect of silymarin in APAP-induced hepatotoxicity in rats. Several biomarkers were assessed and compared: ALT and AST for hepatic function, oxidative stress parameters (NO and MDA), antioxidant parameters (thiols and catalase), inflammatory cytokine (TNF-alpha), MMP-2 and MMP-9 as biomarkers for ECM degradation.

Materials and Methods

Animals. Male albino rats (Wistar-Bratislava) were provided by the Animal Department of the Faculty of Medicine and Pharmacy, Cluj-Napoca. The animals weighing 200-250 mg were kept in polypropylene cages, each group in a separate cage, at a constant temperature (24±2°C) and 60±5% humidity, in a 12/12 h light-dark cycle. Unrestricted access to food (standard pellets from Cantacuzino Institute, Bucharest, Romania) and water was provided. Prior to blood sample collection, the animals were fasted overnight. The experimental protocol was approved by the Ethics Committee of “Iuliu Hatieganu” University of Pharmacy, Cluj-Napoca, Romania (No 16/20.05.2019), and complied with the rules of the European Convention for the Protection of Vertebrate Animals used for Experimental and other Scientific Purposes.

Figure 1.
Figure 1.

The levels of ALT between the APAP group and the control group were significantly different (p<0.01). The same results (p<0.01) were obtained from the comparison of the APAP group with APAP+S, APAP+LCC1 and APAP+LCC2 groups, and for the comparison of different pretreatments (APAP+S with APAP+LCC1 and APAP+LCC2). LCC1 and LCC2 pretreatments yielded also statistically significant differences with p<0.05. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Experimental design. In this study, five groups of 7 rats each were used as follows: Group 1 - control group - intraperitoneal (i.p.) administration of 1 ml saline solution 0.9%; Group 2 - i.p. administration of APAP [a single dose of 250 mg/kg body weight (bw)] after 16 h of fasting (32, 33); Group 3 - APAP+silymarin (silymarin (100 mg/kg bw) was administered i.p. once per day for 5 consecutive days before APAP administration) (34);

Group 4 - APAP+LCC1 - i.p. – 1 mg/100 g bw; Group 5 - APAP+LCC2 - i.p. – 2 mg/100 g bw.

LCC1 and LCC2 pretreatments were performed 30 min before APAP administration.

Substances. Liposomal curcumin was encapsulated in long-circulating liposomes at a concentration of 4.7 mg/ml, using the film hydration method with a lipid molar ratio of 9.5:0.5:1 (1,2-dipalmitoyl-sn-glycero-3-phosphocholine:1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000]-DPPC:PEG-2000-DSPE:CHO) as previously described (35, 36). The proposed formulation had appropriate quality attributes for i.p. administration, such as monodisperse size around 140 nm and zeta potential of about -50 mV. Silymarin and the substances used for biochemical determinations were purchased from Sigma-Aldrich (St. Louis, MO, USA). Quantitative assessment of TNF-α was performed using the ELISA technique according to the manufacturer's instructions (kit purchased from Signosis Inc., Santa Clara, CA, USA). For the quantitative determination of rat matrix metalloproteinases 2 and 9 (MMP-2, MMP-9), we used an ELISA kit according to the manufacturer's protocol (Elabscience Biotechnology Inc., Houston, TX, USA).

Blood collection and assessment of serum markers for oxidative stress/antioxidant status, transaminases, TNF-α, MMP-2 and MMP-9. Blood samples were collected at the end of the experiment (24 h) from the retro-orbital plexus of each animal, under ketamine anesthesia (5 mg/kg bw, i.p. route) (37), and biochemical measurements were used for serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The oxidative stress parameters were measured according to Tsikas for indirect assessment of NOx and according to Janero for MDA determinations (38, 39). Antioxidant parameters, catalase and thiols, were assessed as previously described (40, 41). At the end of the experiment, the animals were euthanized [by ketamine overdose – intramuscular (i.m.) route]. Spectroscopic measurements were performed using a Jasco V-350 UV-VIS spectrophotometer (Jasko International Co, Ltd., Japan) for all biochemical analyses. TNF-α and MMP measurements were made using the ELISA method with an ELISA plate reader (DAS, Rome, Italy).

Histopathological analysis. The livers were collected and fixed in 10% formaldehyde solution. Subsequently, they were paraffin embedded and sectioned at 5 μm, stained with hematoxylin-eosin and examined for histological changes using a light microscope. The graded lesions were subjectively classified as absent, moderate and severe, according to the presence of lesions.

Statistical analysis. The results were expressed as mean±standard deviation for each group. Differences between groups were compared for the degree of oxidative/antioxidant parameters, ALT, AST levels, TNF-α, MMP-2, and MMP-9 levels, using Man-Whitney test. Statistix 10 software was used and differences were considered significant at p<0.05.

Figure 2.
Figure 2.

The levels of AST between the APAP group and the control group were significantly different (p<0.01). The same results (p<0.01) were obtained from the comparison of the APAP group with APAP+S, APAP+LCC1 and APAP+LCC2 groups, and for the comparison of different pretreatments (APAP+S with APAP+LCC1 and APAP+LCC2). LCC1 and LCC2 pretreatments yielded also statistically significant differences with p<0.05. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Figure 3.
Figure 3.

The levels of TNF-α were significantly different between the APAP group and the control group (p<0.01). Similar results (p<0.05) were obtained for the comparison between APAP+S, APAP+LCC1 and APAP+LCC2 groups. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Results

Treatment of rats with APA resulted in hepatoxicity as indicated by the increase, compared to control, in ALT and AST, markers of liver damage, and TNF-α, marker of inflammation (Table I, Figures 1, 2 and 3). In addition, after induction of hepatotoxicity, the oxidative stress parameters (NOx and MDA) were also significantly (Mann-Whitney test p<0.002) increased (Table I, Figures 4 and 5) as well as matrix metalloproteinases MMP-2 and MMP-9 (Table I, Figures 6 and 7). However, a significant decrease in the antioxidant capacity, quantified by thiols and catalase, was observed after APAP administration (Table I, Figures 8 and 9).

Figure 4.
Figure 4.

NOx value showed statistically significant differences in all the comparisons (p<0.001). C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Figure 5.
Figure 5.

The levels of MDA were statistically significantly different at the level of p<0.01, when comparing C with APAP; APAP with APAP+S, APAP+LCC1 and APAP+LCC2, and at the level of p<0.05 when comparing APAP+S with APAP+LCC1 and APAP+LCC2; APAP+LCC1 and APAP+LCC2. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Effect of LCC1 and LCC2 pre-treatment on the levels of transaminases.

Pretreatment of rats with silymarin significantly (p<0.01) decreased ALT and AST levels following APAP administration (Figures 1 and 2). Furthermore, if the animals were pre-treated once with LCC, both ALT and AST levels were significantly (p<0.01) reduced compared with the APAP group (Figures 1 and 2). When comparing the hepatoprotective effect of the combination of silymarin with two liposomal curcumin concentrations, we observed an even more significant (p<0.01) decrease in AST and ALT levels compared to silymarin treatment alone, showing better improvement in hepatic cell function. Both doses of LCC were compared. The lowest values of AST and ALT were obtained with the high dose LCC2 (Figures 1 and 2, p<0.05).

Figure 6.
Figure 6.

The levels of MMP-2 were significantly different between all compared groups (p<0.01) except between the LCC1 pretreatment group and the LCC2 pretreatment group p<0.05. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Figure 7.
Figure 7.

The levels of MMP-9 were statistically significantly different between all compared groups (p<0.01). C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Effect of LCC1 and LCC2 pre-treatment on inflammation. TNF-α level was significantly (p<0.01) decreased is rats pretreated with the combination of silymarin with two doses of LCC (Figure 3), showing the anti-inflammatory effect of both substances during liver damage. Both doses of LCC were more efficient in decreasing inflammation than silymarin (p<0.05), however, the higher dose of LCC2 was more efficient than LCC1 in decreasing the levels of TNF-α (p<0.05) (Figure 3).

Figure 8.
Figure 8.

The levels of thiols, an antioxidant parameter, were statistically significant different at the level of p<0.01 when comparing C with APAP; APAP with APAP+S, APAP+LCC1 and APAP+LCC2, and at the level of p<0.05 when comparing APAP+S with APAP+LCC1 and APAP+LCC2; APAP+LCC1 and APAP+LCC2. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Figure 9.
Figure 9.

The levels of CAT, an antioxidant parameter, were statistically significant different at the levels of p<0.01 when comparing C with APAP; APAP with APAP+S, APAP+LCC1 and APAP+LCC2, and at the level of p<0.05 when comparing APAP+S with APAP+LCC1 and APAP+LCC2; APAP+LCC1 and APAP+LCC2. C: Control; APAP: acetaminophen administration; APAP+S: APAP and sylimarin administration as pretreatment; APAP+LCC1: APAP and pretreatment with 1 mg/100 g bw LCC; APAP+LCC2: APAP and pretreatment with 2 mg/100 g bw LCC.

Oxidative/antioxidant balance after LCC1 and LCC2 treatment. Oxidative stress markers induced by APAP administration were significantly decreased if the animals pre-treated with silymarin or LCC (Figures 4 and 5). The most significant decrease was observed for NOx, with a p<0.001 (Figure 4). Pre-treatment with LCC had a stronger effect than silymarin on reducing oxidative stress, as showed by the significant (p<0.05) decrease in MDA (Figure 5) and even more significant (p<0.001) decrease in NOx (Figure 4). The effect of LCC was dose-dependent and the higher dose had a stronger effect (p<0.05 for MDA and p<0.001 for NOx).

Figure 10.
Figure 10.

Liver group 2, hematoxylin-eosin staining, 40X ob; 1 – zone 1 of the hepatic acinus; 2 – zone 2 of the hepatic acinus; 3 – zone 3 of the hepatic acinus.

The antioxidant capacity was evaluated by measuring thiols and catalase. APAP administration resulted in reduced anti-oxidant capacity, which was reversed upon pre-treatment with silymarin or LCC; thus, significantly increased levels of thiols (p<0.01, Figure 8) and catalase (p<0.01, Figure 9) were observed in the treated groups. LCC pre-treatment reversed the antioxidant capacity to a higher level than silymarin (p<0.05 for thiols and catalase). The higher dose of LCC (LCC2) was the most effective in restoring antioxidant activity (p<0.05 for both thiols and catalase, when compared to LCC1).

Effect of LCC1 and LCC2 pre-treatment on MMP-2 and MMP-9. As shown above APAP treatment resulted in the activation of MMP-2 and MMP-9. This effect was significantly reduced (p<0.01) if the animals were pre-treated with silymarin or LCC (Figures 6 and 7). LCC had a significant (p<0.01) better effect in lowering both MMP-2 and MMP-9 levels than silymarin. The values of MMP-2 and MMP-9 reached those of the control group in the group treated with the highest dose of LCC (LCC2) (Figure 6 with p<0.05 for MMP-2 and Figure 7 with p<0.01 for MMP-9).

Histology assessment. In group 2 of animals, a large number of hepatocytes showed hydropic degeneration. Hepatocytes around the centrilobular venule (zone 3 of the hepatic acinus) were affected in high numbers. In zone 2 of the hepatic acinus, cells in various hydropic degeneration stages were found, among which cells with discrete signs of granular degeneration were interspersed (Figure 10). In group 3 of animals, lesions were less extensive; in zone 2 of the hepatic acinus, the number of hepatocytes with hydropic degeneration was reduced (Figure 11). In group 4 of rats, lesions had a predominantly granular appearance and mainly intersected hepatocytes in zone 2 of the hepatic acinus. In zone 3 of the hepatic acinus, hepatocytes had agranulovacuolar degeneration appearance (Figure 12). In group 5 of animals, the number of cells in zone 3 of the hepatic acinus with granulovacuolar degeneration appearance was further reduced compared to group 4. In zone 2 of the hepatic acinus, the majority of hepatocytes showed discrete granular degeneration and only in some places the degenerative appearance was more pronounced. In zone 1 of the hepatic acinus, degeneration seemed to be at an earlier stage compared to the other zones of the acinus (Figure 13).

Figure 11.
Figure 11.

Liver group 3, hematoxylin-eosin staining, 40X ob; 1 – zone 1 of the hepatic acinus; 2 – zone 2 of the hepatic acinus; 3 – zone 3 of the hepatic acinus.

Discussion

Liposomal curcumin effect on hepatic function and oxidative stress/antioxidant balance in APA-induced hepatotoxicity. Administration of an overdose of APAP resulted in destruction of hepatocytes and elevation of serum ALT and AST levels (Table I; Figures 1 and 2). Transaminase elevations are the most commonly used biomarkers for hepatocyte lesions. Hepatocyte destruction results from depletion of GLUT, a component of the liver antioxidant system, and from increased oxidative stress due to hepatic metabolism of APAP (42). Administration of APAP caused a dose-dependent depletion of hepatic GLUT (that plays a protective role against APAP overdose) (43). APAP is also oxidized by cytochrome P450 to a reactive metabolite, which can cause liver lesions (44). This mechanism leads to mitochondrial oxidative stress, increased mitochondrial membrane permeability and hepatic cell death (45). Mitochondrial oxidative stress is considered to be the main cellular dysfunction in APAP-induced liver injury (46). In our study, the levels of oxidative stress molecules (NOx and MDA) significantly increased after APAP administration (250 mg/kg) (Table II, Figures 4 and 5). Similar results were reported by other authors, thus, the suggested mechanism is APAP-induced activation of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) in hepatocytes (47). Other authors have reported that APAP can induce liver injury via an oxidative stress mechanism caused by increased MDA production (48). Besides GLUT, which has been proven to be depleted by APAP overdose, a decrease in other antioxidant molecules could change the cell oxidant/antioxidant balance. A significant reduction of thiol and catalase levels was obtained after APAP administration (Table II; Figures 8 and 9). Global protein sulfhydryl levels have been reported to decrease significantly starting one hour after APAP overdose, and slowly decreased after 24 h (49). This fact may provide a new insight into studying novel therapeutic molecules that can provide hepatoprotection; one of the mechanisms could be represented by improving the level of thiols. Improving catalase levels in experimental APAP-induced hepatotoxicity could also provide a new approach for hepatoprotection strategies (50). Therefore, modulation of oxidative stress/antioxidant balance could be the target of nutraceuticals used to provide a hepatoprotective effect. One of the most studied natural compounds for its antioxidative properties is curcumin. We have already demonstrated the beneficial effect of orally administered curcumin on hepatic function and on oxidative stress/antioxidant balance in experimental fructose-induced metabolic syndrome in rats (51). In this study, after APAP-induced hepatotoxicity challenge, the elevation in plasma ALT, AST, and oxidative stress molecules was ameliorated by pretreatment with nanoformulation of curcumin (lipososmal curcumin). We also obtained better results with LCC pretreatment compared to silymarin and a dose-dependent effect LCC, where a higher dose (LCC2) had higher efficiency (Table I and II; Figures 1, 2, 3, 4 and 5, 8, 9). The hepatoprotective effect of LCC could be linked to its already proven inhibitory effect on iNOS activity (52) and MDA production (51) as well as its ability to maintain the thiol pool (53) and to improve catalase production (54). In addition to mitochondrial oxidative stress, many other cellular processes, including inflammation, microcirculatory dysfunction and extracellular matrix degradation, have been shown to be involved in the pathogenesis of APAP-induced liver injury (46, 55).

Figure 12.
Figure 12.

Liver group 4, hematoxylin-eosin staining, 10X ob; 1 – zone 1 of the hepatic acinus; 2 – zone 2 of the hepatic acinus; 3 – zone 3 of the hepatic acinus.

Figure 13.
Figure 13.

Liver group 5, hematoxylin-eosin staining, 10X ob; 1 – zone 1 of the hepatic acinus; 2 – zone 2 of the hepatic acinus; 3 – zone 3 of the hepatic acinus.

Liposomal curcumin effect on pro-inflammatory cytokine TNF-α in APAP-induced hepatotoxicity. In APAP hepatotoxicity, increased oxidative stress results in DNA fragmentation followed by an inflammatory reaction and production of pro-inflammatory cytokines (5). One of the most representative pro-inflammatory cytokines associated with hepatic liver injury is TNF-α (56). TNF-α is produced in response to liver injury induced by APAP overdose (57). Activated liver macrophages (Kupffer cells) are the main cells responsible for TNF-α production that further mediates hepatocyte necrosis (58). Our study observations (Table I; Figure 3) are in agreement with other studies that have reported an increase in TNF-α in APAP-induced hepatotoxicity (57, 59, 60). A possible mechanism is the enhancement of an intense inflammatory reaction that includes TNF-α increase through oxidative stress (61). TNF-α and other pro-inflammatory cytokines can initiate the pathological changes following APAP overdose (62), which were also observed in our study (Table I, Figure 3). TNF-α is an important cytokine that can trigger the apoptosis cascade through activation of the caspase-dependent apoptosis mechanism (63). Continuous release of TNF-α during inflammation may rapidly lead to necroptosis and necrosis through an unknown mechanism (62). Multiple studies have demonstrated the role of curcumin in TNF-α modulation (63-65), but not as a liposomal formula. Our study demonstrated the hepatoprotective effect of both LCC concentrations in reducing the serum levels of TNF-α in APAP-induced hepatotoxicity (Table I, Figure 3). Both LCC concentrations were more efficient than silymarin pretreatment (Table I, Figure 3). TNF-α reduction was also demonstrated by oral curcumin administration or by other curcumin nanoformulations (66-68). The liposomal formulation of curcumin can enhance its anti-inflammatory properties through down-regulation of TNF-α due to its higher bioavailability, and a better distribution in therapeutic concentration at the injury site (69).

View this table:
Table II.

Levels of the investigated markers of oxidative stress and antioxidant parameters in each treatment group, expressed as mean and standard deviation.

Liposomal curcumin effect on MMP-2 and MMP-9 in APAP-induced hepatotoxicity. Significantly higher levels of MMP-2 and MMP-9 were observed following APAP administration (Table III; Figures 6 and 7). Both LCC1 and LCC2 reduced the MMP-2 and MMP-9 with LCC2 having the stronger effect, when compared with APAP+S group (Table III; Figures 6 and 7). Increased serum MMP levels have been shown to be associated with hepatocellular damage, contributing to hepatic microcirculation dysfunction and facilitating the arrival of leukocytes at the injury site (70). MMP-2/MMP-9 inhibition has been proven to reduce parenchymal and microvascular injury through minimizing endothelial injury (70). After liver contusion, some authors have reported an enhancement of MMP-2 at 6 h, with a peak at 24 h, then a gradual reduction with normalization of the levels after 7 days (71). ECM degradation by MMP is added to the hepatic microcirculation disturbances caused by NOx excess (72). Intracellular damage induced by APAP overdose is amplified by increased MMP and elicits a robust inflammatory response with the release of pro-inflammatory cytokines that will attract more neutrophils at the injury site (73). After the endothelial injury, accumulated neutrophils together with MMP will contribute to liver injury, but also to liver regeneration after the lesion stimulus ceases (73). It is important to mention that MMPs have an important physiological role in the liver. MMPs are produced by various cells such as Kupffer cells, hepatocytes, cholangiocytes and, in normal amounts, can control inflammation and fibrosis in hepatic tissue (74). Based on these findings and on our results, we consider that the excess of MMP-2 and MMP-9 contributes to the increase of oxidative stress and to the propagation of inflammation, amplifying the hepatic cytolysis process. Similarly to our results, other studies have reported a reduction in liver lesions by down-regulation of MMP production (75-78). Curcumin suppression of MMP expression has also been reported by other studies (79-80), but our findings proved that the liposomal formulation of curcumin can have beneficial effects in improving oxidative stress/antioxidant balance and inflammation in hepatotoxicity induced by APAP in rats. To our knowledge, this is the first report regarding the hepatoprotective effects of a liposomal formulation of curcumin.

View this table:
Table III.

Metalloproteinase levels in each treatment group.

Conclusion

Despite all studies performed so far, there are very few therapy choices for liver injuries. This study provides a new perspective for therapeutic strategies to alleviate hepatic lesions related to APAP overdose. Our results provide evidence that liposomal curcumin can exert a protective effect against APAP-induced hepatotoxicity by reducing NOx and MDA production, improving thiol and catalase levels, and reducing the serum concentration of TNF-α, MMP-2 and MMP-9. Therefore, liposomal curcumin formula may be a promising therapy for APAP-induced liver injury, with beneficial effects on associated oxidative stress and inflammation.

Acknowledgements

The Authors would like to thank Dr. Alina Porfire and Lucia Tefas for the liposomal curcumin preparation, Ana Uifalean for biochemical assessments, and Mirel Molnar for helping in rat confinement and for collecting the blood samples.

Footnotes

  • Authors' Contributions

    Bulboacă Adriana Elena – study concept and design, analysis and interpretation of the data, writing the first draft of the manuscript; Dogaru Gabriela – study concept, organization of manuscript, review and critique; Gheban Dan – histological analysis; Boarescu Paul Mihai – organization of the manuscript, analysis and interpretation of the data; Rus Vasile – manuscript review and critique; Festila Dana – organization of the manuscript, analysis and interpretation of the data; Sitar-Taut Adela-Viviana – statistical analysis; Stănescu Ioana – organization of manuscript, review and critique.

  • This article is freely accessible online.

  • Conflicts of Interest

    The Authors declare no conflicts of interest regarding this study.

  • Received October 30, 2019.
  • Revision received December 2, 2019.
  • Accepted December 3, 2019.

References

  1. ↵
    1. Subramanya SB,
    2. Venkataraman B,
    3. Meeran MFN,
    4. Goyal SN,
    5. Patil CR,
    6. Ojha S
    : Therapeutic potential of plants and plant derived phytochemicals against acetaminophen-induced liver injury. Int J Mol Sci 19(12): E3776, 2018. PMID: 30486484. DOI: 10.3390/ijms19123776
    OpenUrl
  2. ↵
    1. Athersuch TJ,
    2. Antoine DJ,
    3. Boobis AR,
    4. Coen M,
    5. Daly AK,
    6. Possamai L,
    7. Nicholson JK,
    8. Wilson ID
    : Paracetamol metabolism, hepatotoxicity, biomarkers and therapeutic interventions: a perspective. Toxicol Res (Camb) 7(3): 347-357, 2018. PMID: 30090586. DOI: 10.1039/c7tx00340d
    OpenUrl
  3. ↵
    1. Almazroo OA,
    2. Miah MK,
    3. Venkataramanan R
    : Drug metabolism in the liver. Clin Liver Dis 21(1): 1-20, 2017. PMID: 27842765. DOI: 10.1016/j.cld.2016.08.001
    OpenUrlCrossRefPubMed
  4. ↵
    1. Bunchorntavakul C,
    2. Reddy KR
    : Acetaminophen-related hepatotoxicity. Clin Liver Dis 17(4): 587-607, 2013. PMID: 24099020. DOI: 10.1016/j.cld.2013.07.005
    OpenUrlCrossRefPubMed
  5. ↵
    1. Jaeschke H,
    2. Duan L,
    3. Akakpo JY,
    4. Farhood A,
    5. Ramachandran A
    : The role of apoptosis in acetaminophen hepatotoxicity. Food Chem Toxicol 118: 709-718, 2018. PMID: 29920288. DOI: 10.1016/j.fct.2018.06.025
    OpenUrl
  6. ↵
    1. Du K,
    2. Ramachandran A,
    3. McGill MR,
    4. Mansouri A,
    5. Asselah T,
    6. Farhood A,
    7. Woolbright BL,
    8. Ding WX,
    9. Jaeschke H
    : Induction of mitochondrial biogenesis protects against acetaminophen hepatotoxicity. Food Chem Toxicol 108: 339-350, 2017. PMID: 28827156. DOI: 10.1016/j.fct.2017.08.020
    OpenUrl
  7. ↵
    1. Ramachandran A,
    2. Visschers RGJ,
    3. Duan L,
    4. Akakpo JY,
    5. Jaeschke H
    : Mitochondrial dysfunction as a mechanism of drug-induced hepatotoxicity: current understanding and future perspectives. J Clin Transl Res 4(1): 75-100, 2018. PMID: 30873497.
    OpenUrl
  8. ↵
    1. Demine S,
    2. Renard P,
    3. Arnould T
    : Mitochondrial Uncoupling: A key controller of biological processes in physiology and diseases. Cells 8(8): E795, 2019. PMID: 31366145. DOI: 10.3390/cells8080795
    OpenUrl
  9. ↵
    1. McGill MR,
    2. Jaeschke H
    : Metabolism and disposition of acetaminophen: recent advances in relation to hepatotoxicity and diagnosis. Pharm Res 30(9): 2174-2187, 2013. PMID: 23462933. DOI: 10.1007/s11095-013-1007-6
    OpenUrlCrossRefPubMed
  10. ↵
    1. Guengerich FP,
    2. Liebler DC
    : Enzymatic activation of chemicals to toxic metabolites. Crit Rev Toxicol 14(3): 259-307, 1985. PMID: 3926381. DOI: 10.3109/10408448509037460
    OpenUrlCrossRefPubMed
  11. ↵
    1. Fadda L,
    2. Ali HM,
    3. Aldrees GA,
    4. Alquraishi NM
    : Nano ubiquinone: Promising candidate for treatment of renal toxicity induced by over dose of paracetamol. Toxicol Rep 6: 712-717, 2019. PMID: 31384564. DOI: 10.1016/j.toxrep.2019.07.008
    OpenUrl
  12. ↵
    1. Gröber U
    : Mitochondrial toxicity of drugs. Med Monatsschr Pharm 235(12): 445-56, 2012. PMID: 23367660.
    OpenUrl
  13. ↵
    1. Attia H,
    2. Al-Rasheed N,
    3. Mohamad R,
    4. Al-Rasheed N,
    5. Al-Amin M
    : The antifibrotic and fibrolytic properties of date fruit extract via modulation of genotoxicity, tissue-inhibitor of metalloproteinases and nuclear factor-kappa B pathway in a rat model of hepatotoxicity. BMC Complement Altern Med 16(1): 414, 2016. PMID: 27776513. DOI: 10.1186/s12906-016-1388-2
    OpenUrl
  14. ↵
    1. Parks WC,
    2. Wilson CL,
    3. Lopez-Boado YS
    : Matrix metalloproteinases as modulators of inflammation and innate immunity. Nat Rev Immunol 4: 617-629, 2004. PMID: 15286728. DOI: 10.1038/nri1418
    OpenUrlCrossRefPubMed
  15. ↵
    1. Duarte S,
    2. Baber J,
    3. Fujii T,
    4. Coito AJ
    : Matrix metalloproteinases in liver injury, repair and fibrosis. Matrix Biol 44-46: 147-156, 2015. PMID: 25599939. DOI: 10.1016/j.matbio.2015.01.004
    OpenUrlCrossRef
  16. ↵
    1. Murthy A,
    2. Shao YW,
    3. Defamie V,
    4. Wedeles C,
    5. Smookler D,
    6. Khokha R
    : Stromal TIMP3 regulates liver lymphocyte populations and provides protection against Th1 T cell-driven autoimmune hepatitis. J Immunol 188: 2876-2883, 2012. PMID: 22323541. DOI: 10.4049/jimmunol.1102199
    OpenUrlAbstract/FREE Full Text
  17. ↵
    1. Hamada T,
    2. Fondevila C,
    3. Busuttil RW,
    4. Coito AJ
    : Metalloproteinase-9 deficiency protects against hepatic ischemia/reperfusion injury. Hepatology 47(1): 186-198, 2008. PMID: 17880014. DOI: 10.1002/hep.21922
    OpenUrlCrossRefPubMed
  18. ↵
    1. Kato H,
    2. Kuriyama N,
    3. Duarte S,
    4. Clavien PA,
    5. Busuttil RW,
    6. Coito AJ
    : MMP-9 deficiency shelters endothelial PECAM-1 expression and enhances regeneration of steatotic livers after ischemia and reperfusion injury. J Hepatol 60(5): 1032-1039, 2014. PMID: 24412604. DOI: 10.1016/j.jhep.2013.12.022
    OpenUrlCrossRefPubMed
  19. ↵
    1. Shehzad A,
    2. Qureshi M,
    3. Anwar MN,
    4. Lee YS
    : Multifunctional curcumin mediate multitherapeutic effects. J Food Sci 82(9): 2006-2015, 2017. PMID: 28771714. DOI: 10.1111/1750-3841.13793
    OpenUrl
  20. ↵
    1. Sreejayan N,
    2. Rao MN
    : Free radical scavenging activity of curcuminoids. Arzneimittelforschung 46: 169-171, 1996. PMID: 8720307.
    OpenUrlPubMed
  21. ↵
    1. Yu H,
    2. Li J,
    3. Shi K,
    4. Huang Q
    : Structure of modified ε-polylysine micelles and their application in improving cellular antioxidant activity of curcuminoids. Food Funct 2: 373-380, 2011. PMID: 21894323. DOI: 10.1039/c1fo10053j
    OpenUrlPubMed
  22. ↵
    1. Jurenka JS
    : Anti-inflammatory properties of curcumin, a major constituent of Curcuma longa: a review of preclinical and clinical research. Altern Med Rev 14: 141-153, 2009. PMID: 19594223.
    OpenUrlPubMed
  23. ↵
    1. Bulboacă A,
    2. D Bolboacă S,
    3. Suci S
    : Protective effect of curcumin in fructose-induced metabolic syndrome and in streptozotocin-induced diabetes in rats. Iran J Basic Med Sci 19(6): 585-593, 2016. PMID: 27482338.
    OpenUrl
    1. Boarescu PM,
    2. Boarescu I,
    3. Bocșan IC,
    4. Pop RM,
    5. Gheban D,
    6. Bulboacă AE,
    7. Nicula C,
    8. Râjnoveanu RM,
    9. Bolboacă SD
    : Curcumin nanoparticles protect against isoproterenol induced myocardial infarction by alleviating myocardial tissue oxidative stress, electrocardiogram, and biological changes. Molecules 24(15): E2802, 2019. PMID: 31374848. DOI: 10.3390/molecules24152802
    OpenUrl
  25. ↵
    1. Boarescu PM,
    2. Chirilă I,
    3. Bulboacă AE,
    4. Bocșan IC,
    5. Pop RM,
    6. Gheban D,
    7. Bolboacă SD
    : Effects of curcumin nanoparticles in isoproterenol-induced myocardial infarction. Oxid Med Cell Longev 4: 1-13, 2019. DOI: 10.1155/2019/7847142
    OpenUrl
  26. ↵
    1. Gera M,
    2. Sharma N,
    3. Ghosh M,
    4. Huynh DL,
    5. Lee SJ,
    6. Min T,
    7. Kwon T,
    8. Jeong DK
    : Nanoformulations of curcumin: an emerging paradigm for improved remedial application. Oncotarget 8(39): 66680-66698, 2017. PMID: 29029547. DOI: 10.18632/oncotarget.19164
    OpenUrl
  27. ↵
    1. Hamano N,
    2. Böttger R,
    3. Lee SE,
    4. Yang Y,
    5. Kulkarni JA,
    6. Ip S,
    7. Cullis PR,
    8. Li SD
    : Robust microfluidic technology and a new lipid composition for fabrication of curcumin-loaded liposomes: Effect on the anticancer activity and safety of cisplatin. Mol Pharm 16(9): 3957-3967, 2019. PMID: 31381352. DOI: 10.1021/acs.molpharmaceut.9b00583
    OpenUrl
  28. ↵
    1. Wang XY,
    2. Ishida T,
    3. Ichihara M,
    4. Kiwada H
    : Influence of the physicochemical properties of liposomes on the accelerated blood clearance phenomenon in rats. J Control Release 104(1): 91-102, 2005. PMID: 15866337. DOI: 10.1016/j.jconrel.2005.01.008
    OpenUrlCrossRefPubMed
    1. Feng T,
    2. Wei Y,
    3. Lee RJ,
    4. Zhao L
    : Liposomal curcumin and its application in cancer. Int J Nanomedicine 12: 6027-6044, 2017. PMID: 28860764. DOI: 10.2147/IJN.S132434
    OpenUrl
  30. ↵
    1. Bulboacă AE,
    2. Porfire AS,
    3. Tefas LR,
    4. Boarescu PM,
    5. Bolboacă SD,
    6. Stănescu IC,
    7. Bulboacă AC,
    8. Dogaru G
    : Liposomal curcumin is better than curcumin to alleviate complications in experimental diabetic mellitus. Molecules 24(5): E846, 2019. PMID: 30818888. DOI: 10.3390/molecules24050846
    OpenUrl
  31. ↵
    1. Zhong S,
    2. Fan Y,
    3. Yan Q,
    4. Fan X,
    5. Wu B,
    6. Han Y,
    7. Zhang Y,
    8. Chen Y,
    9. Zhang H,
    10. Niu J
    : The therapeutic effect of silymarin in the treatment of nonalcoholic fatty disease: A meta-analysis (PRISMA) of randomized control trials. Medicine (Baltimore) 96(49): e9061, 2017. PMID: 29245314. DOI: 10.1097/MD.0000000000009061
    OpenUrl
  32. ↵
    1. Uchida NS,
    2. Silva-Filho SE,
    3. Cardia GFE,
    4. Cremer E,
    5. Silva-Comar FMS,
    6. Silva EL,
    7. Bersani-Amado CA,
    8. Cuman RKN
    : Hepatoprotective effect of citral on acetaminophen-induced liver toxicity in mice. Evid Based Complement Alternat Med 2017: 1796209, 2017. PMID: 28717379. DOI: 10.1155/2017/1796209
    OpenUrl
  33. ↵
    1. Nafady AM,
    2. Ahmed OB,
    3. Ghafeer HH
    : Scanning and transmission electron microscopy of the cells forming the hepatic sinusoidal wall of rat in acetaminophen and Escherichia coli endotoxin-induced hepatotoxicity. J Microsc Ultrastruct 5(1): 21-27, 2017. PMID: 30023233. DOI: 10.1016/j.jmau.2016.04.003
    OpenUrl
  34. ↵
    1. Kwon KW,
    2. Nam Y,
    3. Choi WS,
    4. Kim TW,
    5. Kim GM,
    6. Sohn UD
    : Hepatoprotective effect of sodium hydrosulfide on hepatic encephalopathy in rats. Korean J Physiol Pharmacol 23(4): 263-270, 2019. PMID: 31297010. DOI: 10.4196/kjpp.2019.23.4.263
    OpenUrl
  35. ↵
    1. Tefas LR,
    2. Sylvester B,
    3. Tomuta I,
    4. Sesarman A,
    5. Licarete E,
    6. Banciu M,
    7. Porfire A
    : Development of antiproliferative long-circulating liposomes co-encapsulating doxorubicin and curcumin, through the use of a quality-by-design approach. Drug Des Devel Ther 11: 1605-1621, 2017. PMID: 28579758. DOI: 10.2147/DDDT.S129008
    OpenUrl
  36. ↵
    1. Bulboacă AE,
    2. Bolboacă SD,
    3. Stănescu IC,
    4. Sfrângeu CA,
    5. Porfire A,
    6. Tefas L,
    7. Bulboacă AC
    : The effect of intravenous administration of liposomal curcumin in addition to sumatriptan treatment in an experimental migraine model in rats. Int J Nanomed 13: 3093-3103, 2018. PMID: 29872296. DOI: 10.2147/IJN.S162087
    OpenUrl
  37. ↵
    1. Melo A,
    2. Leite-Almeida H,
    3. Ferreira C,
    4. Sousa N,
    5. Pêgo JM
    : Exposure to ketamine anesthesia affects rat impulsive behavior. Front Behav Neurosci 10: 226, 2016. PMID: 27932959. DOI: 10.3389/fnbeh.2016.00226
    OpenUrl
  38. ↵
    1. Tsikas D
    : Analysis of nitrite and nitrate in biological fluids by assays based on the Griess reaction: appraisal of the Griess reaction in the L-arginine/nitric oxide area of research. J Chromatogr B Analyt Technol Biomed Life Sci 851(1-2): 51-70, 2007. PMID: 16950667. DOI: 10.1016/j.jchromb.2006.07.054
    OpenUrlCrossRefPubMed
  39. ↵
    1. Janero DR
    : Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 9(6): 515-540, 1990. PMID: 2079232. DOI: 10.1016/0891-5849(90)90131-2
    OpenUrlCrossRefPubMed
  40. ↵
    1. Sies H
    1. Aebi H
    : Catalase in vitro. In: Oxygen radicals in biological systems. Vol. 105 of Methods in enzymology. Sies H (ed.). Cambridge, Academic Press, Mass., pp. 121-126, 1984.
  41. ↵
    1. Hu ML
    : Measurement of protein thiol groups and glutathuine in plasma. Methods Enzymol 233: 380-384, 1994. PMID: 8015473. DOI: 10.1016/s0076-6879(94)33044-1
    OpenUrlCrossRefPubMed
  42. ↵
    1. Mazer M,
    2. Perrone J
    : Acetaminophen-induced nephrotoxicity: pathophysiology, clinical manifestations, and management. J Med Toxicol 4(1): 2-6, 2008. PMID: 18338302. DOI: 10.1007/bf03160941
    OpenUrlCrossRefPubMed
  43. ↵
    1. Mitchell JR,
    2. Jollow DJ,
    3. Potter WZ,
    4. Gillette JR,
    5. Brodie BB
    : Acetaminophen-induced hepatic necrosis. IV. Protective role of glutathione. J Pharmacol Exp Ther 187(1): 211-217, 1973. PMID: 4746329.
    OpenUrlAbstract/FREE Full Text
  44. ↵
    1. Caro AA,
    2. Cederbaum AI
    : Inhibition of cyp2e1 catalytic activity in vitro by s-adenosyl-l-methionine. Biochem Pharmacol 69: 1081-1093, 2005. PMID: 15763544. DOI: 10.1016/j.bcp.2005.01.007
    OpenUrlCrossRefPubMed
  45. ↵
    1. Ramachandran A,
    2. McGill MR,
    3. Xie Y,
    4. Ni HM,
    5. Ding WX,
    6. Jaeschke H
    : The receptor interacting protein kinase 3 is a critical early mediator of acetaminophen-induced hepatocyte necrosis in mice. Hepatology 58(6): 2099-2108, 2013. PMID: 23744808. DOI: 10.1002/hep.26547
    OpenUrlCrossRefPubMed
  46. ↵
    1. Yan M,
    2. Huo Y,
    3. Yin S,
    4. Hu H
    : Mechanisms of acetaminophen-induced liver injury and its implications for therapeutic interventions. Redox Biol 17: 274-283, 2018. PMID: 29753208. DOI: 10.1016/j.redox.2018.04.019
    OpenUrl
  47. ↵
    1. Trettin A,
    2. Böhmer A,
    3. Suchy MT,
    4. Probst I,
    5. Staerk U,
    6. Stichtenoth DO,
    7. Frölich JC,
    8. Tsikas D
    : Effects of paracetamol on NOS, COX, and CYP activity and on oxidative stress in healthy male subjects, rat hepatocytes, and recombinant NOS. Oxid Med Cell Longev 2014: 212576, 2014. DOI: 10.1155/2014/212576.
    OpenUrl
  48. ↵
    1. Hu JN,
    2. Liu Z,
    3. Wang Z,
    4. Li XD,
    5. Zhang LX,
    6. Li W,
    7. Wang YP
    : Ameliorative effects and possible molecular mechanism of action of black ginseng (Panax ginseng) on acetaminophen-mediated liver injury. Molecules 22(4): E664, 2017. PMID: 28430162. DOI: 10.3390/molecules22040664
    OpenUrl
  49. ↵
    1. Yang X,
    2. Greenhaw J,
    3. Shi Q,
    4. Roberts DW,
    5. Hinson JA,
    6. Muskhelishvili L,
    7. Davis K,
    8. Salminen WF
    : Mouse liver protein sulfhydryl depletion after acetaminophen exposure. J Pharmacol Exp Ther 344(1): 286-294, 2013. PMID: 23093024. DOI: 10.1124/jpet.112.199067
    OpenUrlAbstract/FREE Full Text
  50. ↵
    1. Zakaria ZA,
    2. Kamisan FH,
    3. Omar MH,
    4. Mahmood ND,
    5. Othman F,
    6. Abdul Hamid SS,
    7. Abdullah MNH
    : Methanol extract of Dicranopteris linearis L. leaves impedes acetaminophen-induced liver intoxication partly by enhancing the endogenous antioxidant system BMC Complement Altern Med 17(1): 271, 2017. PMID: 28521788. DOI: 10.1186/s12906-017-1781-5
    OpenUrl
  51. ↵
    1. Bulboacă AE,
    2. Bolboacă SD,
    3. Bulboacă AC,
    4. Porfire AS,
    5. Tefas LR,
    6. Suciu ȘM,
    7. Dogaru G,
    8. Stănescu IC
    : Liposomal curcumin enhances the effect of naproxen in a rat model of migraine. Med Sci Monit 25: 5087-5097, 2019. PMID: 31287810. DOI: 10.12659/MSM.915607
    OpenUrl
  52. ↵
    1. Somchit M,
    2. Changtam C,
    3. Kimseng R,
    4. Utaipan T,
    5. Lertcanawanichakul M,
    6. Suksamrarn A,
    7. Chunglok W
    : Demethoxycurcumin from Curcuma longa rhizome suppresses iNOS induction in an in vitro inflamed human intestinal mucosa model. Asian Pac J Cancer Prev 15(4): 1807-1810, 2014. PMID: 24641413. DOI: 10.7314/apjcp.2014.15.4.1807
    OpenUrlPubMed
  53. ↵
    1. Zhang J,
    2. Bai KW,
    3. He J,
    4. Niu Y,
    5. Lu Y,
    6. Zhang L,
    7. Wang T
    : Curcumin attenuates hepatic mitochondrial dysfunction through the maintenance of thiol pool, inhibition of mtDNA damage, and stimulation of the mitochondrial thioredoxin system in heat-stressed broilers. J Anim Sci Apr 96(3): 867-879, 2018. PMID: 29566233. DOI: 10.1093/jas/sky009
    OpenUrl
  54. ↵
    1. Heritage M,
    2. Jaskowski L,
    3. Bridle K,
    4. Campbell C,
    5. Briskey D,
    6. Britton L,
    7. Fletcher L,
    8. Vitetta L,
    9. Subramaniam VN,
    10. Crawford D
    : Combination curcumin and vitamin E treatment attenuates diet-induced steatosis in Hfe−/− mice. World J Gastrointest Pathophysiol 8(2): 67-76, 2017. PMID: 28573069. DOI: 10.4291/wjgp.v8.i2.67
    OpenUrl
  55. ↵
    1. Bhushan B,
    2. Edwards,
    3. Desai A,
    4. Michalopoulos GK,
    5. Apte U
    : Liver-specific deletion of integrin-linked kinase in mice attenuates hepatotoxicity and improves liver regeneration after acetaminophen overdose. Gene Expr 17(1): 35-45, 2016. PMID: 27125733. DOI: 10.3727/105221616X691578
    OpenUrl
  56. ↵
    1. Hammam O,
    2. Mahmoud O,
    3. Zahran M,
    4. Sayed A,
    5. Salama R,
    6. Hosny K,
    7. Farghly A
    : A possible role for TNF-α in coordinating inflammation and angiogenesis in chronic liver disease and hepatocellular carcinoma. Gastrointest Cancer Res 6(4): 107-114, 2013. PMID: 24147158.
    OpenUrlPubMed
  57. ↵
    1. Blazka ME,
    2. Elwell MR,
    3. Holladay SD,
    4. Wilson RE,
    5. Luster MI
    : Histopathology of acetaminophen-induced liver changes: role of interleukin 1 alpha and tumor necrosis factor alpha. Toxicol Pathol 24(2): 181-189, 1996. PMID: 8992608. DOI: 10.1177/019262339602400206
    OpenUrlCrossRefPubMed
  58. ↵
    1. Mochida S,
    2. Ogata I,
    3. Ohta Y,
    4. Yamada S,
    5. Fujiwara K
    : In situ evaluation of the stimulatory state of hepatic macrophages based on their ability to produce superoxide anions in rats. J Pathol 158(1): 67-71, 1989. PMID: 2547045. DOI: 10.1002/path.1711580113
    OpenUrlCrossRefPubMed
  59. ↵
    1. Gardner CR,
    2. Hankey P,
    3. Mishin V,
    4. Francis M,
    5. Yu S,
    6. Laskin JD,
    7. Laskin DL
    : Regulation of alternative macrophage activation in the liver following acetaminophen intoxication by stem cell-derived tyrosine kinase. Toxicol Appl Pharmacol 262(2): 139-148, 2012. PMID: 22575169. DOI: 10.1016/j.taap.2012.04.027
    OpenUrlCrossRefPubMed
  60. ↵
    1. Chiu H,
    2. Gardner CR,
    3. Dambach DM,
    4. Brittingham JA,
    5. Durham SK,
    6. Laskin JD,
    7. Laskin DL
    : Role of p55 tumor necrosis factor receptor 1 in acetaminophen-induced antioxidant defense. Am J Physiol Gastrointest Liver Physiol 285(5): G959-66, 2003. PMID: 12842828. DOI: 10.1152/ajpgi.00219.2003
    OpenUrlCrossRefPubMed
  61. ↵
    1. Arai KI,
    2. Lee F,
    3. Miyajima A,
    4. Miyatake S,
    5. Arai N,
    6. Yokota T
    : Cytokines: coordinators of immune and inflammatory responses. Annu Rev Biochem 59: 783-836, 1990. PMID: 1695833. DOI: 10.1146/annurev.bi.59.070190.004031
    OpenUrlCrossRefPubMed
  62. ↵
    1. Yan T,
    2. Wang H,
    3. Zhao M,
    4. Yagai T,
    5. Chai Y,
    6. Krausz KW,
    7. Xie C,
    8. Cheng X,
    9. Zhang J,
    10. Che Y,
    11. Li F,
    12. Wu Y,
    13. Brocker CN,
    14. Gonzalez FJ,
    15. Wang G,
    16. Hao H
    : Glycyrrhizin protects against acetaminophen-induced acute liver injury via alleviating Tumor Necrosis Factor α-mediated apoptosis. Drug Metab Dispos 44(5): 720-731, 2016. PMID: 26965985. DOI: 10.1124/dmd.116.069419
    OpenUrlAbstract/FREE Full Text
  63. ↵
    1. Kelany ME,
    2. Hakami TM,
    3. Omar AH
    : Curcumin improves the metabolic syndrome in high-fructose-diet-fed rats: role of TNF-α, NF-κB, and oxidative stress. Can J Physiol Pharmacol 95(2): 140-150, 2017. PMID: 27901349. DOI: 10.1139/cjpp-2016-0152
    OpenUrl
    1. Chen Q,
    2. Wang T,
    3. Li J,
    4. Wang S,
    5. Qiu F,
    6. Yu H,
    7. Zhang Y,
    8. Wang T
    : Effects of natural products on fructose-induced nonalcoholic fatty liver disease (NAFLD). Nutrients 9(2): E96, 2017. PMID: 28146130. DOI: 10.3390/nu9020096
    OpenUrl
  65. ↵
    1. Mohammadi A,
    2. Blesso CN,
    3. Barreto GE,
    4. Banach M,
    5. Majeed M,
    6. Sahebkar A
    : Macrophage plasticity, polarization and function in response to curcumin, a diet-derived polyphenol, as an immunomodulatory agent. J Nutr Biochem 66: 1-16, 2019. PMID: 30660832. DOI: 10.1016/j.jnutbio.2018.12.005
    OpenUrl
  66. ↵
    1. Mouzaoui S,
    2. Rahim I,
    3. Djerdjouri B
    : Aminoguanidine and curcumin attenuated tumor necrosis factor (TNF)-α-induced oxidative stress, colitis and hepatotoxicity in mice. Int Immunopharmacol 12(1): 302-311, 2012. PMID: 22036766. DOI: 10.1016/j.intimp.2011.10.010
    OpenUrlCrossRefPubMed
    1. Trivedi MK,
    2. Mondal SC,
    3. Gangwar M,
    4. Jana S
    : Immunomodulatory potential of nanocurcumin-based formulation. Inflammopharmacology 25(6): 609-619, 2017. PMID: 28921388. DOI: 10.1007/s10787-017-0395-3
    OpenUrl
  68. ↵
    1. Jazayeri-Tehrani SA,
    2. Rezayat SM,
    3. Mansouri S,
    4. Qorbani M,
    5. Alavian SM,
    6. Daneshi-Maskooni M,
    7. Hosseinzadeh-Attar MJ
    : Efficacy of nanocurcumin supplementation on insulin resistance, lipids, inflammatory factors and nesfatin among obese patients with non-alcoholic fatty liver disease (NAFLD): a trial protocol. BMJ Open 7(7): e016914, 2017. PMID: 28698350. DOI: 10.1136/bmjopen-2017-016914
    OpenUrlAbstract/FREE Full Text
  69. ↵
    1. Gupta SC,
    2. Patchva S,
    3. Aggarwal BB
    : Therapeutic roles of curcumin: lessons learned from clinical trials. AAPS J 15(1): 195-218, 2013. PMID: 23143785. DOI: 10.1208/s12248-012-9432-8
    OpenUrlCrossRefPubMed
  70. ↵
    1. Ito Y,
    2. Abril ER,
    3. Bethea NW,
    4. McCuskey RS
    : Inhibition of matrix metalloproteinases minimizes hepatic microvascular injury in response to acetaminophen in mice. Toxicol Sci 83(1): 190-196, 2005. PMID: 15456921. DOI: 10.1093/toxsci/kfh291
    OpenUrlCrossRefPubMed
  71. ↵
    1. Wang CL,
    2. Xia ZX,
    3. Zhang GH,
    4. You JB,
    5. Yu H,
    6. Wang LL,
    7. Zhang MZ,
    8. Yang HB,
    9. Gu JP,
    10. Jin YK
    : Time-dependent protein expressions of MMP-2 and MMP-9 in liver contusion rats after impact. Fa Yi Xue Za Zhi 33(6): 581-586, 2017. PMID: 29441762. DOI: 10.3969/j.issn.1004-5619.2017.06.002
    OpenUrl
  72. ↵
    1. Ito Y,
    2. Abril ER,
    3. Bethea NW,
    4. McCuskey RS
    : Role of nitric oxide in hepatic microvascular injury elicited by acetaminophen in mice. Am J Physiol Gastrointest Liver Physiol 286(1): G60-67, 2004. PMID: 29441762. DOI: 10.3969/j.issn.1004-5619.2017.06.002
    OpenUrl
  73. ↵
    1. Alvarenga DM,
    2. Mattos MS,
    3. Lopes ME,
    4. Marchesi SC,
    5. Araújo AM,
    6. Nakagaki BN,
    7. Santos MM,
    8. David BA,
    9. De Souza VA,
    10. Carvalho É,
    11. Sousa Pereira RV,
    12. Marques PE,
    13. Mafra K,
    14. de Castro Oliveira HM,
    15. de Miranda CDM,
    16. Diniz AB,
    17. de Oliveira THC,
    18. Teixeira MM,
    19. Rezende RM,
    20. Antunes MM,
    21. Menezes GB
    : Paradoxical role of matrix metalloproteinases in liver injury and regeneration after sterile acute hepatic failure. Cells 7(12): pii: E247, 2018. PMID: 30563238. DOI: 10.3390/cells7120247
  74. ↵
    1. Zbodakova O,
    2. Chalupsky K,
    3. Tureckova J,
    4. Sedlacek R
    : Metalloproteinases in liver fibrosis: current insights. Metalloproteinases Med 2017(4): 25-35, 2017. DOI:10.2147/MNM.S124363
    OpenUrl
  75. ↵
    1. Viappiani S,
    2. Sariahmetoglu M,
    3. Schulz R
    : The role of matrix metalloproteinase inhibitors in ischemia-reperfusion injury in the liver. Curr Pharm Des 12(23): 2923-34, 2006. PMID: 16918422. DOI: 10.2174/138161206777947560
    OpenUrlCrossRefPubMed
    1. Cursio R,
    2. Mari B,
    3. Louis K,
    4. Rostagno P,
    5. Saint-Paul MC,
    6. Giudicelli J,
    7. Bottero V,
    8. Anglard P,
    9. Yiotakis A,
    10. Dive V,
    11. Gugenheim J,
    12. Auberger P
    : Rat liver injury after normothermic ischemia is prevented by a phosphinic matrix metalloproteinase inhibitor. FASEB J 16(1): 93-95 2002. PMID: 11709491. DOI: 10.1096/fj.01-0279fje
    OpenUrlCrossRefPubMed
    1. Defamie V,
    2. Laurens M,
    3. Patrono D,
    4. Devel L,
    5. Brault A,
    6. Saint-Paul MC,
    7. Yiotakis A,
    8. Barbry P,
    9. Gugenheim J,
    10. Crenesse D,
    11. Dive V,
    12. Huet PM,
    13. Mari B
    : Matrix metalloproteinase inhibition protects rat livers from prolonged cold ischemia-warm reperfusion injury. Hepatology 47(1): 177-185, 2008. PMID: 18008367. DOI: 10.1002/hep.21929
    OpenUrlPubMed
  78. ↵
    1. Al-Olayan EM,
    2. El-Khadragy MF,
    3. Aref AM,
    4. Othman MS,
    5. Kassab RB,
    6. Abdel Moneim AE
    : The potential protective effect of Physalis peruviana L. against carbon tetrachloride-induced hepatotoxicity in rats is mediated by suppression of oxidative stress and downregulation of MMP-9 expression. Oxid Med Cell Longev 2014: 381413, 2014. PMID: 24876910. DOI: 10.1155/2014/381413
    OpenUrlPubMed
  79. ↵
    1. Morsy MA,
    2. Abdalla AM,
    3. Mahmoud AM,
    4. Abdelwahab SA,
    5. Mahmoud ME
    : Protective effects of curcumin, α-lipoic acid, and N-acetylcysteine against carbon tetrachloride-induced liver fibrosis in rats. J Physiol Biochem 68(1): 29-35, 2012. PMID: 21986891. DOI: 10.1007/s13105-011-0116-0
    OpenUrlPubMed
  80. ↵
    1. Zhong Y,
    2. Feng J,
    3. Li J,
    4. Fan Z
    : Curcumin prevents lipopolysaccharide-induced matrix metalloproteinase-2 activity via the Ras/MEK1/2 signaling pathway in rat vascular smooth muscle cells. Mol Med Rep 16(4): 4315-4319, 2017. PMID: 28731157. DOI: 10.3892/mmr.2017.7037
    OpenUrl
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Effect of Liposomal Curcumin on Acetaminophen Hepatotoxicity by Down-regulation of Oxidative Stress and Matrix Metalloproteinases
GABRIELA DOGARU, ADRIANA ELENA BULBOACA, DAN GHEBAN, PAUL MIHAI BOARESCU, VASILE RUS, DANA FESTILA, ADELA-VIVIANA SITAR-TAUT, IOANA STANESCU
In Vivo Mar 2020, 34 (2) 569-582; DOI: 10.21873/invivo.11809
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