Methotrexate-induced Liver Injury Research Articles

Evaluation of concurrent use of Vitamin C and Niclosamide against methotrexate-induced liver injury in mice

Objective: To examine the effect of using vitamin C and niclosamide together on liver damage caused by methotrexate. Method: The study was conducted at the Pharmacology Department and the Iraqi Center for Cancer and Medical Genetics Research, College of Medicine, Mustansiriya University, Baghdad, Iraq, from November 2020 to July 2021, and comprised albino mice who were randomly assigned to 5 groups. Group 1 comprised controls, groups 2 to 5 was received methotrexate, niclosamide 70mg/kg/day, vitamin C 100mg/kg/day, and a combination of niclosamide and vitamin C, respectively. Mice in groups 3, 4 and 5 also received an intraperitoneal injection of methotrexate 20mg/kg to induce hepatotoxicity. After 48 hours of the injection, the mice were sacrificed under chloroform anaesthesia. Cardiac blood samples were drawn for biochemical examination. The liver, after being washed, was divided into two parts; one part was taken for histological examination, and the other was preserved in formalin 10% for histopathological analyses. Data was analysed using SPSS 16. Results: Of the 35 mice, there were 7(20%) in each of the 5 groups. The overall age ranged between 9-12 weeks and weight between 18-38gm. The results show significant hepatoprotection ( p-value <0.05) produced by both niclosamide and Vitamin C separately, reflected by a decrease in ALP, ALT, and LDH, while the combination of (niclosamide and Vitamin C) showed no additive effect (p>0.05) on enhancement of liver function. Conclusion: Niclosamide alone was found to be superior than in combination with vitamin C for treating methotrexate-induced liver damage. Key Words: Methotrexate, Chloroform, Niclosamide, Intraperitoneal, Liver, Formaldehyde

Monotropein mitigates methotrexate-induced liver injury by activating autophagy and inhibiting ferroptosis

Methotrexate (MTX) is effective for the therapy of cancer, lupus erythematosus, rheumatoid arthritis, psoriasis and other diseases. However, the hepatotoxicity of MTX often leads to withdrawal and discontinuation of drugs, and then limits its clinical application. Monotropein (MON), an iridoid glycosides distributing in many edible and medicinal plants, has been exhibited to prevent liver and kedney injury induced by chemicals and related diseases. The current study aimed to assess the potential beneficial effects of MON in mitigating MTX-induced liver injury together with delineating the implicated mechanisms. A MTX-induced liver injury model in mice and BRL 3A cells were developed to explore the hepatoprotection of MON. GEO and WGCNA analysis were conducted to predict the possible mechanism of MTX on liver injury. The results demonstrated that MON can decrease the serum levels of AST and ALT, increase the activities of antioxidant SOD and GSH, and improve the morphological alteration of liver tissue, and reduce the accumulation of lipids in hepatocytes and liver tissue of mice treated with MTX. Moreover, our results also exhibited that MON can activate autophagy as assessed by autophagosome formation, expression of protein related with autophagy, and inhibit ferroptosis as evaluated by mitochondrial integrity, Fe2+level and key protein expression changes, and CQ (inhibitor of autophagy) and RSL-3 (the activator of ferroptosis) reversed the effects of MON on autophagy and ferroptosis of hepatocytes injured by MTX. Therefore, MON attenuates liver injury of MTX by activating autophagy and inhibiting ferroptosis. These findings provide new insights into MON as a potential healthy ingredient in functional food.

Morinda officinalis iridoid glycosides alleviate methotrexate-induced liver injury in CIA rats by increasing liver autophagy and improving lipid metabolism homeostasis

Ethnopharmacological relevanceMorinda officinalis How. is a commonly used traditional Chinese herb with the pharmacological properties of tonifying liver and kidney, and enhancing bone and muscle. Iridoid glycosides are the predominant components of this plant, including monotropein, asperuloside, deacetylasperuloside and deacetylasperulosidic acid with their contents reaching more than 2%. Methotrexate (MTX) is the drug of choice for the treatment of rheumatoid arthritis (RA), but liver injury induced by MTX limits its wider use for RA. Morindaofficinalis iridoid glycoside (MOIG) is reported as having anti-RA and hepatoprotective effects, but the exact efficacy on MTX-induced liver injury and the underlying molecular mechanism remain unclear. AimTo elucidate the mitigating effect of MOIG against liver injury in RA rats treated with MTX, and explore the possible mechanism. Materials and methodsThe effect and mechanism of MOIG were investigated in Wistar rats with collagen-induced arthritis (CIA) which were then treated with MTX, and MTX-induced hepatocyte injury in vitro. Network pharmacological and transcriptomic analyses were conducted to predict the possible mechanisms of MOIG in mitigating MTX-induced liver injury, and lipidomic analysis was performed to further verify the regulatory effects of MOIG on lipid metabolism. BRL-3A hepatocytes were used to evaluate the regulatory effects of MOIG against MTX-associated liver injury. ResultsMOIG treatment enhanced the anti-RA effect of MTX, and mitigated oxidative damage, inflammation and apoptosis of liver tissues in CIA rats treated with MTX. Network pharmacological and transcriptomic analyses demonstrated that MOIG attenuated liver injury by regulating autophagy and lipid metabolism. The result of lipidomic analysis showed that MOIG reversed the disturbance of lipid metabolism of the liver tissue in CIA rats after MTX treatment. In addition, MOIG also inhibited the apoptosis, reduced the levels of lactate dehydrogenase (LDH), aspartate aminotransferase (ALT) and alanine aminotransferase (AST), regulated oxidative stress, and increased the formation of autophagosome and translocation of LC3 in the nucleus and expression of autophagy regulatory genes Beclin-1, ATG5, LC3Ⅱ, ATG7 and ATG12 in hepatocytes subjected to MTX damage. ConclusionOur findings demonstrated that MOIG could ameliorate MTX-induced liver injury in the treatment of RA through increasing hepatocyte autophagy and improving lipid metabolism homeostasis.

Liver Elastography for the Detection of Methotrexate-Induced Liver Injury: A Retrospective Study

Background: Liver biopsy, the gold standard for monitoring of methotrexate-induced liver injury, is associated with significant morbidity and mortality. Transient elastography (TE) has been used as a non-invasive alternative to detect liver stiffness. Aim: To assess the utility of TE in detecting liver fibrosis in patients with methotrexate use. Methods: A retrospective chart review was performed for 35 patients referred to the liver clinic for evaluation of suspected methotrexate-induced liver injury. Demographic, clinical, histopathological, and elastographic data were collected and interpreted. Liver stiffness measurement (LSM) and controlled attenuation parameter were recorded from TE results. Results: Thirty-five patients with a mean age of 58 years, including 23 females (66%), were included. The median LSM by TE was 10.8 kPa and the median controlled attenuation parameter was 303 dB/m. A total of 12 out of 35 patients (34%) had evidence of clinical and pathological advanced fibrosis. Using a cut-off elastography value of 10 kPa, the TE yielded 92% sensitivity and 93% negative predictive value for ruling out methotrexate-induced advanced liver fibrosis. Using a higher LSM cut-off point of kPa ≥15.0, specificity was calculated at 87% and positive predictive value at 80%. Area under the receiver operating characteristic curve was 0.80 (95% confidence interval). Conclusion: FibroScan® (Echosens, Paris, France) has a high sensitivity and specificity for kPa 10 and 15, respectively, for detecting advanced liver fibrosis in patients on methotrexate.

Chemistry of Bairui granules and its mechanisms in the protective effect against methotrexate-induced liver injury.

Bairui granules (BRKL), a traditional Chinese medicine preparation, possess a range of pharmacological effects. However, its impact on methotrexate (MTX)-induced liver damage remains unexplored. The present work focused on investigating the potential protection of BRKL on MTX-induced liver damage, along with its potential active ingredients and underlying mechanisms. We evaluated the hepatoprotective activities of BRKL in liver-damaged Wistar rats induced by intraperitoneal MTX injection, observing the liver's morphological and pathological features. Additionally, we measured serum ALT, AST, and LDH levels using kits. Ultra High-Performance Liquid Chromatography-Q-Exactive Orbitrap Mass Spectrometry (UHPLC-Q-Exactive Orbitrap MS) analyzed BRKL composition, and network pharmacology strategy predicted and analyzed BRKL's targets and pathways. Thereafter, we conducted molecular docking for analyzing affinity of bioactive ingredients for targets with Autodock. At last, results were verified through in vitro experiments. The animal experiments revealed the significant protection of BRKL against MTX-mediated rat liver damage. A total of 64 major chemical constituents were identified in BRKL by UHPLC-Q-Exactive Orbitrap MS. We then applied the network-based pharmacological strategy to clarify BRKL's molecular mechanism on liver damage based on the identified components. The targets EGFR, SRC, PIK3R1, AKT1, and ESR1, as well as compounds isorhamnetin 3,7-O-diglucoside, β-ecdysone, chrysoeriol, apigenin, and diosmetin, may play pivotal roles in treating MTX-mediated liver damage. According to our in vitro experiments, isorhamnetin 3,7-O-diglucoside may exert its liver-protective effect via AKT/NF-κB pathway. BRKL protected against MTX-mediated liver injury, and the bioactive ingredients, key pathways, and liver injury-related molecular targets have been identified. These findings provide new insights into using BRKL in treating liver damage and propose a feasible approach to exploring phytomedicine's chemical and pharmacological foundation.

Potential Hepatoprotective Effects of Chamaecyparis lawsoniana against Methotrexate-Induced Liver Injury: Integrated Phytochemical Profiling, Target Network Analysis, and Experimental Validation.

Methotrexate (MTX) therapy encounters significant limitations due to the significant concern of drug-induced liver injury (DILI), which poses a significant challenge to its usage. To mitigate the deleterious effects of MTX on hepatic function, researchers have explored plant sources to discover potential hepatoprotective agents. This study investigated the hepatoprotective effects of the ethanolic extract derived from the aerial parts of Chamaecyparis lawsoniana (CLAE) against DILI, specifically focusing on MTX-induced hepatotoxicity. UPLC-ESI-MS/MS was used to identify 61 compounds in CLAE, with 31 potential bioactive compounds determined through pharmacokinetic analysis. Network pharmacology analysis revealed 195 potential DILI targets for the bioactive compounds, including TP53, IL6, TNF, HSP90AA1, EGFR, IL1B, BCL2, and CASP3 as top targets. In vivo experiments conducted on rats with acute MTX-hepatotoxicity revealed that administering CLAE orally at 200 and 400 mg/kg/day for ten days dose-dependently improved liver function, attenuated hepatic oxidative stress, inflammation, and apoptosis, and reversed the disarrayed hepatic histological features induced by MTX. In general, the findings of the present study provide evidence in favor of the hepatoprotective capabilities of CLAE in DILI, thereby justifying the need for additional preclinical and clinical investigations.

Methotrexate-Induced Liver Injury Is Associated with Oxidative Stress, Impaired Mitochondrial Respiration, and Endoplasmic Reticulum Stress In Vitro

Low-dose methotrexate (MTX) is a standard therapy for rheumatoid arthritis due to its low cost and efficacy. Despite these benefits, MTX has been reported to cause chronic drug-induced liver injury, namely liver fibrosis. The hallmark of liver fibrosis is excessive scarring of liver tissue, triggered by hepatocellular injury and subsequent activation of hepatic stellate cells (HSCs). However, little is known about the precise mechanisms through which MTX causes hepatocellular damage and activates HSCs. Here, we investigated the mechanisms leading to hepatocyte injury in HepaRG and used immortalized stellate cells (hTERT-HSC) to elucidate the mechanisms leading to HSC activation by exposing mono- and co-cultures of HepaRG and hTERT-HSC to MTX. The results showed that at least two mechanisms are involved in MTX-induced toxicity in HepaRG: (i) oxidative stress through depletion of glutathione (GSH) and (ii) impairment of cellular respiration in a GSH-independent manner. Furthermore, we measured increased levels of endoplasmic reticulum (ER) stress in activated HSC following MTX treatment. In conclusion, we established a human-relevant in vitro model to gain mechanistical insights into MTX-induced hepatotoxicity, linked oxidative stress in HepaRG to a GSH-dependent and -independent pathway, and hypothesize that not only oxidative stress in hepatocytes but also ER stress in HSCs contribute to MTX-induced activation of HSCs.

Losartan prevents methotrexate-induced liver and lung injury in rats via targeting PPAR-γ/TGF-β1/SMAD3 and Nrf2/redox signaling

Losartan prevents methotrexate-induced liver and lung injury in rats via targeting PPAR-γ/TGF-β1/SMAD3 and Nrf2/redox signaling

Astaxanthin effect on apoptotic biomarkers in methotrexate-induced liver injury

Methotrexate is used in the treatment of cancer, psoriasis, rheumatoid arthritis and several other disorders. It has a hepatotoxic potential side effect. Patients who have no access to alternative medications face a serious
 
 challenge as a result. The current study aimed to assess the apoptotic potential of methotrexate on liver cells and evaluate the hepatoprotective activity of the potent antioxidant astaxanthin, by downregulation of apoptotic biomarkers caspase 9 and caspase 3.
 A model of methotrexate-induced liver toxicity was employed on male rats. Thirty-six rats were divided into six groups; a negative control group, methotrexate induction group given (20 mg/kg) on day 13, three groups pretreated with astaxanthin in ascending doses (50, 75 and 100 mg/kg) for 14 days before methotrexate, and a conventional therapy group pretreated with silymarin (200mg/kg).
 The use of methotrexate significantly increased liver tissue caspase 9 and caspase 3 compared to the negative control. On the other side, astaxanthin used in all three doses significantly normalized these biomarkers. This study revealed that since astaxanthin significantly decreased caspase 9 and caspase 3 that are involved in the apoptotic pathway, it could be used as pretreatment in patients treated with methotrexate to alleviate its hepatotoxicity.

The protective effect of alfa lipoic acid against methotrexate induced acute liver injury in mice

Methotrexate induced liver injury is one of major problem resulted from it is use Previous study found that one of mechanism responsible for liver injury caused by methotrexate is oxidative stress, Alfa lipoic acid is an antioxidant and antiinflammatory dietary supplement. Aim of study is to investigate the protective effect of low and high dose of Alfa lipoic acid on methotrexate induced acute liver injury. Twenty eight male adult albino mice were divided into 4 group and in each group 7 mice, group 1 were administered distilled water and sodium bicarbonate orally by oral gavage for ten days and injected intrperitoneally normal saline on the tenth day only, group 2 were injected MTX intraperitoneally on the tenth day only, group 3 were administered orally 60 mg/kg Alfa lipoic acid orally for ten days and injected methotrexate intraperitoneally on the tenth day ,group 4 were administered orally 120mg/kg Alfa lipoic acid for ten days and injected methotrexate on the tenth day , after two days of the last treatment , the mice were anesthesized and blood was withdrawn for biochemical assessment of liver function , then sacrifing the mice and the liver was dissected out for liver tissue oxidative stress biomarker determination and histopathological study. The results show that serum level of liver enzyme and lipid peroxidation was increase ,reduced glutathione and superoxide dismutase was decrease in methotrexate group, group pretreated with 60mg/kg alfa lipoic acid was showed an improvement of liver enzyme elevation and decrease level of MDA , group pretreated with 120 mg/kg alfa lipoic acid was showed more improvement in liver enzyme elevation and induce more reduction of malondialdehyde and increase level of reduced glutathione. Conclusion: pretreatment by utilizing high dose of alfa lipoic acid lead to produce more significant protection against methotrexate induced liver injury.

Modulation of Gut Microbiota by Magnesium Isoglycyrrhizinate Mediates Enhancement of Intestinal Barrier Function and Amelioration of Methotrexate-Induced Liver Injury.

BackgroundThe gut–liver axis plays a crucial role in various liver diseases. Therefore, targeting this crosstalk may provide a new treatment strategy for liver diseases. However, the exact mechanism underlying this crosstalk and its impact on drug-induced liver injury (DILI) requires clarification.AimThis study aimed to investigate the potential mechanism and therapeutic effect of MgIG on MTX-induced liver injury, which is associated with the gut–liver axis and gut microbiota.MethodsAn MTX-induced liver injury model was generated after 20-mg/kg/3d MTX application for 30 days. Meanwhile, the treatment group was treated with 40-mg/kg MgIG daily. Histological examination, aminotransferase, and aspartate aminotransferase enzyme levels were estimated to evaluate liver function. Immune cells infiltration and inflammatory cytokines were detected to indicate inflammation levels. Colon histological score, intestinal barrier leakage, and expression of tight junctions were employed to assess the intestinal injury. Bacterial translocation was observed using fluorescent in situ hybridisation, colony-forming unit counting, and lipopolysaccharide detection. Alterations in gut microbial composition were analysed using 16s rDNA sequencing and relative quantitative polymerase chain reaction. Short-chain-fatty-acids and lactic acid concentrations were then utilized to validate changes in metabolites of specific bacteria. Lactobacillus sp. supplement and fecal microbiota transplantation were used to evaluate gut microbiota contribution.ResultsMTX-induced intestinal and liver injuries were significantly alleviated using MgIG treatment. Bacterial translocation resulting from the intestinal barrier disruption was considered a crucial cause of MTX-induced liver injury and the therapeutic target of MgIG. Moreover, MgIG was speculated to have changed the gut microbial composition by up-regulating probiotic Lactobacillus and down-regulating Muribaculaceae, thereby remodelling the intestinal barrier and inhibiting bacterial translocation.ConclusionThe MTX-induced intestinal barrier was protected owing to MgIG administration, which reshaped the gut microbial composition and inhibited bacterial translocation into the liver, thus attenuating MTX-related DILI.

Mechanistic Insights on the In Vitro Antibacterial Activity and In Vivo Hepatoprotective Effects of Salvinia auriculata Aubl against Methotrexate-Induced Liver Injury.

Methotrexate (MTX) is widely used in the treatment of numerous malignancies; however, its use is associated with marked hepatotoxicity. Herein, we assessed the possible hepatoprotective effects of Salvinia auriculata methanol extract (SAME) against MTX-induced hepatotoxicity and elucidated the possible fundamental mechanisms that mediated such protective effects for the first time. Forty mice were randomly allocated into five groups (eight/group). Control saline, MTX, and MTX groups were pre-treated with SAME 10, 20, and 30 mg/kg. The results revealed that MTX caused a considerable increase in blood transaminase and lactate dehydrogenase levels, oxidative stress, significant activation of the Nod-like receptor-3 (NLPR3)/caspase-1 inflammasome axis, and its downstream inflammatory cytokines interleukin-1β (IL-1β) and interleukin-18 (IL-18). MTX also down-regulated nuclear factor erythroid 2-related factor 2 (Nrf2) expression. Additionally, it increased the immunostaining of nuclear factor kappa-B (NF-κB) and downstream inflammatory mediators. Furthermore, the hepatic cellular apoptosis was dramatically up-regulated in the MTX group. On the contrary, prior treatment with SAME significantly improved biochemical, histopathological, immunohistochemical alterations caused by MTX in a dose-dependent manner. The antibacterial activity of SAME has also been investigated against Acinetobacter baumannii clinical isolates. LC-ESI-MS/MS contributed to the authentication of the studied plant and identified 24 active constituents that can be accountable for the SAME-exhibited effects. Thus, our findings reveal new evidence of the hepatoprotective and antibacterial properties of SAME that need further future investigation.

Hepatoprotective effect of Olea europaea L. seeds extracts against methotrexate induced liver injury in mice

The present study aimed to determine the hepatoprotective effect of different extracts of Olea europaea L. seeds in mice induced with MTX-induced liver injury and find out the most effective extract. Olea europaea L. (olive) seeds powder was extracted with three solvents with different polarities in a sequential manner (chloroform, methanol, and 50% aqueous ethanol) using the cold method. 48 male albino mice were divided into six groups. Mice were pretreated orally with silymarin (standard) and

Hepatoprotective Effects of Alpha-Lipoic Acid, Vitamin C Alone, or in Combination On Methotrexate-Induced Liver Injury

Background: Different mechanisms have been attributed to methotrexate (MTX)-induced liver injury, one of which is related to MTX generation of free radicals and inducing the disturbance of oxidative stress biomarker. Alpha-lipoic acid (ALA) is a strong antioxidant dietary supplement and has an anti-inflammatory effect. Vitamin C is an antioxidant and might produce an anti-inflammatory action through its antioxidant effect. The Aim of the Current Study: To evaluate the effect of ALA and Vitamin C single or concurrent use of them on MTX-induced liver injury. Materials and Methods: Thirty-five adult male albino mice were divided into five groups: Group 1 were administered distilled water and sodium bicarbonate orally by oral gavage for 10 days and injected normal saline intraperitoneally (ip) in the 10th day, Group 2 were injected MTX ip on the 10th day only, Group 3 were administered 100 mg/kg Vitamin C orally for 10 days and injected MTX ip on the 10th day, Group 4 were administered ALA 60 mg/kg orally for 10 days and injected MTX ip on the 10th day, Group 5 were administered both ALA and Vitamin C orally and injected MTX ip on the 10th day. After 2 days of the last treatment, the animal was anesthesized and the blood was withdrawn to be used for biochemical assessment of liver functions and liver tissue was dissected out to be used for oxidative stress biomarker determination and histopathological study. Results: MTX group showed an increase in serum level of alanine aminotransferase, alkaline phosphatase, and lactate dehydrogenase, an increase liver tissue malondialdehyde (MDA), and a reduction of tissue glutathione and superoxide dismutase (SOD), while the treatment groups showed an improvement of liver enzyme elevation and decreased lipid peroxidation MDA and increased SOD. Conclusion: Pretreatment by using ALA alone or vitamin alone provides comparable improvement of liver histology and liver injury while combined regimen did not provide more significant improvement of liver injury than using each one alone.

Attenuation of Liver Injury via Bicyclol–Bovine Serum Albumin Nanopreparation using a Green Synthetic Approach

Bicyclol (BIC) is a traditional antihepatitis drug that is used to treat chronic hepatitis B infections by improving liver function and reducing transaminase levels. Because the extensive use of BIC for treating liver injury is limited by its poor bioavailability, the course of treatment with oral BIC lasted for >6 months. This study aimed to develop a nano‐BIC injection to improve its bioavailability and anti‐hepatitis efficacy. We used a green synthetic approach to prepare BIC‐bovine serum albumin (BSA) nanoparticles (BIC‐NPs). Moderate protein denaturation is required to release free thiol groups in the intramolecular reactions of BSA, which may form an intermolecular disulfide network to stabilize the BIC–BSA nanoassembly. Our results showed that the administration of BIC‐NPs in rats resulted in a 2.42‐fold increase in drug concentration in plasma. Further, nano‐BIC injection showed high bioavailability and rapidly achieved therapeutic concentrations. BIC can restore mitochondrial function by scavenging reactive oxygen species. Moreover, we observed that 70 nm NPs can accumulate in the liver, and the nano‐BIC injection protected mice from methotrexate‐induced liver injury. This study provides an improved strategy for developing a liver‐protecting agent to meet various clinical needs of patients.

A Risk Scoring Model for High-Dose Methotrexate-Induced Liver Injury in Children With Acute Lymphoblastic Leukemia Based on Gene Polymorphism Study.

A study on 70 acute lymphoblastic leukemia (ALL) children (age ≤16 years) treated with high-dose methotrexate (HD-MTX) in Sichuan Provincial People’s Hospital was conducted. The aim of the study was to establish a risk-scoring model to predict HD-MTX-induced liver injury, considering gene polymorphisms’ effects. Data screening was performed through t-test, chi-square test, and ridge regression, and six predictors were identified: age, MTRR_AA, MTRR_AG, SLCO1B1_11045879_CC, albumin_1 day before MTX administration, and IBIL_1 day before MTX administration (p < 0.1). Then, the risk-scoring model was established by ridge regression and evaluated the prediction performance. In a training cohort (n = 49), the area under the curve (AUC) was 0.76, and metrics including accuracy, precision, sensitivity, specificity, positive predictive value, and negative predictive value were promising (0.86, 0.81, 0.76, 0.91, 0.81, 0.88, respectively). In a test cohort (n = 21), the AUC was 0.62 and negative predictive value was 0.80; other evaluation metrics were not satisfactory, possibly due to the limited sample size. Ultimately, the risk scores were stratified into three groups based on their distributions: low- (≤48), medium- (49–89), and high-risk (>89) groups. This study could provide knowledge for the prediction of HD-MTX-induced liver injury and reference for the clinical medication.

Human placental extract ameliorates methotrexate-induced hepatotoxicity in rats via regulating antioxidative and anti-inflammatory responses

PurposeMethotrexate (MTX) induces hepatotoxicity, limiting its clinical efficacy as a widely known chemotherapy drug. In the current study, we examined the protective effect of human placenta extract (HPE) against MTX-induced liver damage in rats, as well as its ability to regulate antioxidative and anti-inflammatory liver responses.MethodsMale rats were orally administered MTX at a daily dose of 5 mg/kg-body-weight in the presence or absence of HPE (10.08 mg/kg) for 2 weeks. We measured the biological effects of MTX and HPE on the levels of liver enzymes, lipid profile, lipid peroxidation, oxidative stress biomarkers, and cytokines [tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10)]. In addition, histological examination and histopathological scoring of liver tissues were performed.ResultsMTX-treated rats showed significantly increased (p < 0.001) liver enzyme levels for aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total bilirubin, total cholesterol, and triglyceride levels. However, HPE supplementation in MTX-treated rats significantly decreased (p < 0.001) these elevated levels. HPE supplementation also significantly reduced the oxidative stress biomarker malondialdehyde (MDA), reversed the reduction in glutathione (GSH), and markedly increased the antioxidant enzyme activities of catalase (CAT) and superoxide dismutase (SOD) in the livers of MTX-treated rats. Furthermore, HPE supplementation significantly decreased the MTX-elevated levels of the pro-inflammatory cytokines TNF-α, IL-6, and IL-10. Histopathological examinations showed that MTX produced severe cellular damage and inflammatory lesions in liver tissues, while treatment with HPE improved hepatic histologic architecture.ConclusionHPE has the ability to ameliorate methotrexate-induced liver injury in rats by mechanisms that include boosting antioxidative responses and down-regulating MDA and pro-inflammatory cytokine production.

Comparative Study between Effect of Niclosamide and Vitamin C on Methotrexate-Induced Liver Injury in Mice

Background: Niclosamide is an old drug used before in the treatment of tapeworms, its main mechanism of action is through the amelioration of mitochondrial-free radical generation. It has been thought that free radical generation has an important role in methotrexate-induced liver injury. Vitamin C, an antioxidant agent, has an essential role in the methotrexate-induced oxidative stress (OS) pathways in mice hepatocytes. Aim: The aim of the study was to evaluate the effect of pretreatment with Vitamin C in different doses and niclosamide in different doses on liver injury, when we induced liver injury with methotrexate. Materials and Methods: Forty-two albino mice were divided equally into six groups, the first group was considered as “the control group,” which received a normal saline solution, the second group was considered as “Methotrexate group,” third and fourth groups were orally pretreated with “niclosamide at a dose (70 mg/kg/day) and (140 mg/kg/day)” respectively, fifth and sixth groups were orally pretreated with “Vitamin C at a dose (100 mg/kg/day) and (200 mg/kg/day),” respectively, all groups, except the control group, were injected with “Methotrexate (20 mg/kg)” intraperitoneally on the 10th day, to induced hepatotoxicity and assessed the effect of the pretreatment with these medications on OS biomarker and histopathological alteration that induced by methotrexate-induced hepatotoxicity. Results: It was found that pretreatment with niclosamide 70 mg/kg/day and 140 mg/kg/day and Vitamin C 100 mg/kg/day and 200 mg/kg/day, in mice injected with methotrexate, led to a decrease in “liver function tests, OS parameters, as well as improvement in liver tissue.” There was an improvement in “serum alanine aminotransferase, alkaline phosphatase, malondialdehyde” in addition to an improvement in “histological appearance,” but it was noted that pretreatment with niclosamide gives a better improvement in “liver function, OS, and liver tissue.” Conclusion: Niclosamide is better than Vitamin C in protecting the hepatocytes against methotrexate-induced liver injury, also niclosamide and Vitamin C have a dose-dependent protecting effect against methotrexate-induced liver injury.

Effects of Omega-3 and Vitamin C on Methotrexate-Induced Liver Injury

Context: Methotrexate (MTX)-induced liver injury is a serious side effect characterized by the increased level of hepatic biomarkers and resulted in acute liver failure. Omega 3 and Vitamin C act as antioxidant that participate in the fighting of free radicals generation during the inflammatory process. Aims: To evaluate the effect of omega 3 and Vitamin C on hepatotoxicity induced by MTX. Settings and Design: 42 (Swiss albino mice) used and divided into six groups (7 mice each): First: Maintained with normal saline, second: Received a single dose injection of MTX (20 mg/kg, intraperitoneally), third: Pretreated with omega 3 100 mg/kg, fourth: Pretreated with omega 3 200 mg/kg, fifth: Pretreated with Vitamin C 100 mg/kg, sixth: Pretreated with Vitamin C 200 mg/kg, then these group injected with MTX on day 10. Subjects and Methods: MTX as 50 mg injection. Omega 3 as capsule 1000 mg. Vitamin C as powder 1000 mg. Assessment of liver enzymes (alanine aminotransferase [ALT], aspartate aminotransferase [AST], and alkaline phosphatase [ALP]) made using automated computering device (Flexor–EL80) provider by Vitalab (South Africa). Assessment of oxidative stress markers (malondialdehyde [MDA], superoxide dismutase [SOD], reduced glutathione [GSH]) and lactate dehydrogenase (LDH) made by using competitive ELISA kits using (ELISA microplate Humareader). Results: This study showed a significant increase in the liver enzymes (ALT, AST, ALP, and LDH) as well oxidative stress markers (MDA, SOD, and GSH) with severe changes in the histopathological findings (severe inflammatory cell necrosis) among group injected with MTX as compared with control group and illustrated improvement in serum level of ALT, ALP, LDH, MDA, SOD and reduced GSH; besides improved histopathological findings (mild and moderate changes) for a group of mice pretreated with omega 3 and Vitamin C. Conclusions: This study concluded that pretreatment with omega 3 (which was strong antioxidant supplement) and Vitamin C (which was dose-dependent manner with beneficial antioxidant action) exert more hepatoprotective effect against oxidative tissue damage induced by MTX.

Glabridin ameliorates methotrexate-induced liver injury via attenuation of oxidative stress, inflammation, and apoptosis

Despite unprecedented advances in modern medicine, no safe and effective drug is available to date for oral administration to combat drug-induced liver injury, which is a vital concern nowadays. The present study deals with the hepatoprotective effect of pure glabridin, a key phytoconstituent from Glycyrrhiza glabra with mechanistic investigations using an in-vivo methotrexate-induced liver injury model as there is no such precedent. The study was performed in the Swiss mice model where a single dose of methotrexate (40 mg/kg) was given on the 7th day through an intraperitoneal route to induce hepatotoxicity, and glabridin as a test compound was administered orally for eleven consecutive days at 10 to 40 mg/kg. Glabridin markedly improved serum biochemical parameters (SGPT, SGOT), proinflammatory cytokine (TNF-α) level, oxidative stress markers (MDA, GSH, SOD, CAT) as compared to methotrexate alone. Alterations in methotrexate-induced liver architecture were considerably prevented by glabridin treatment as suggested by liver histopathological examination and SEM investigation. Glabridin substantially prevented methotrexate-induced down-regulation of Nrf2, & activation of NF-κB, and caused up-regulation of BAX at different dose levels. Overall, glabridin is found to protect methotrexate-induced hepatotoxicity by improving important factors for oxidative stress, inflammation, and apoptosis.