Induced Liver Toxicity Research Articles

Hepatoprotective effect of berberine against methotrexate induced liver toxicity in rats

Hepatotoxicity is one of the major side effects of methotrexate (MTX), which restricts the clinical use of this drug. Berberine (BBR) is a natural compound with multiple pharmacological activities such as antioxidant, antiapoptotic and anti-inflammatory effects. In this study, the effect of BBR on MTX-induced hepatotoxicity was studied. A total number of 28 male Wistar rats were randomly divided into four experimental groups. Rats were pretreated with BBR orally with dose of 100mg/kg for 10 consecutive days and MTX (20mg/kg, intraperitoneally) was administrated on the 9th day. Then on day 11, blood samples were collected to determine serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase (ALP). The extracted livers were used for histological examination, biochemical assays and real time PCR studies. Malondialdehyde (MDA), glutathione (GSH), protein carbonyl (PC), nitric oxide (NO) levels, catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and myeloperoxidase (MPO) activities were assessed in hepatic tissue. In addition, the expression of SOD and PGx was measured using real-time PCR method in hepatic tissue. Results showed that MTX administration significantly increases AST, ALT and ALP levels (all p<0.001). It also, increases MDA, PC, NO levels and MPO activity (p<0.001, p<0.01, p<0.05 and p<0.01 respectively). Moreover, MTX decreases hepatic GSH level, SOD, GPx and CAT activities (all p<0.001). Pre-treatment with BBR for 10days prevented some of these changes. Serum levels of AST and ALT decreased (all p<0.001). Hepatic MDA level decreased (p<0.001) and GSH level as well as GPx activity increased (p<0.05 and p<0.01 respectively). Our results indicated that BBR might be useful for prevention of the hepatotoxicity induced by MTX via ameliorative effects on biochemical and oxidative stress indices.

HEPATOPROTECTIVE EFFECT OF CHENOPODIUM QUINOA SEED AGAINST CCL4-INDUCED LIVER TOXICITY IN SWISS ALBINO MALE MICE

Objective: To study the effect of Quinoa seed on carbon tetrachloride (CCL4)-induced liver toxicity in Swiss albino male mice.Methods: Swiss albino male mice were divided into three groups: Group 1 served as control group; Group 2 served as hepatotoxic group (CCl4 treated); and Group 3 served as combination of Quinoa seed powder (20 mg/kg) + CCl4-treated group. The effect of these groups on liver tissue was studied after three different periods of 4, 8, and 12 weeks. Liver marker enzyme level of serum glutamic-oxaloacetic transaminase, serum glutamic-pyruvic transaminase, serum alkaline phosphatase and liver antioxidant enzyme level of lipid peroxidation, superoxide dismutase, and reduced glutathione were measured. Further, histopathological analysis of liver was also conducted.Results: The results showed that the treatment with Quinoa seed powder significantly reduced CCl4-induced elevated serum level of hepatic enzymes and restored the level of liver antioxidant enzymes toward the normal levels, which was also confirmed by histopathological studies.Conclusion: Results of the present study concluded that the Quinoa seed (Chenopodium quinoa) showed hepatoprotective effect against CCl4-induced liver damage in Swiss albino male mice.

Low dose administration of Bisphenol A induces liver toxicity in adult rats

BackgroundBisphenol A (BPA) is one of the most widely used chemicals, often used in epoxy resins, health products and colors. This study aims to investigate the effect of various doses of BPA on hepatotoxicity in rats. MethodThis experimental study was conducted using 20 male adult Wistar rats older than 2 months weighing 150–200 g. (5, 25 and 125 μg/kg) BPA was administered by gavage for 35 consecutive days. The animals were weighed at the beginning and the end of the experiment. The level of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase were determined using colorimetric method. The liver tissue was kept in the freezer at −80 °C for histological studies. FindingThe body weight of rats receiving BPA decreased significantly compared to control group and this weight loss was more evident at doses of 25 and 125 μg/kg (p < 0.001). Results of the study demonstrated that the level of ALP and AST decreases significantly (p < 0.001 and p < 0.05, respectively), while the level of ALT did not change. The results that BPA significantly decreased Beta-2 protein and increased Gama protein serum levels in rats (p < 0.01). ConclusionResults of this study demonstrated that BPA increase gamma globulin protein levels and decreases the level of alkaline phosphatase, aspartate aminotransferase and serum protein β2 and causes weight loss in rats after treatment. This research also demonstrated that the toxic effect of BPA on liver is induced by oxidative stress.

Hepatoprotective Effect of Curcumin and Capsaicin against Lipopolysaccharide Induced Liver Damage in Mice

Objective: The present study was undertaken to evaluate the possible ameliorative role of curcumin, capsaicin and their combination against lipopolysaccharide (LPS) induced hepatic toxicity in mice. Methods: Animals were distributed into five experimental groups: Normal control, vehicle control, curcumin, capsaicin and combined curcumin and capsaicin treatment groups respectively, for 7 days prior to LPS induced liver toxicity (3 mg/kg b.w. in saline). Hepatoprotective effect of individual and combined spice principles were evidenced by the measurement of serum marker enzyme activities such as, SGPT, ALP and TB and it was further confirmed by histopathological observation of liver tissue section. Results: The administration of LPS increased serum nonspecific enzymes (SGOT; 174.2±3.79 IU/L, SGPT; 124.0±3.14 IU/L, ALP; 320.15±3.88 IU/L and total bilirubin level; 2.32±1.23 mg/dL), however dietary curcumin and capsaicin decreased the activities of these non–specific serum enzymes including total bilirubin indicating amelioration of the severe LPS induced hepatotoxicity, while the combined spice principles were more significant as shown by the levels of enzymes activities SGOT; 89.9±1.39 IU/L, SGPT; 85.9±1.83 IU/L, ALP; 138.4±2.05 IU/L including total bilirubin level; 0.86±0.03 mg/dL. Conclusion: Dietary curcumin and capsaicin individually are protective to LPS induced hepatotoxicity, the beneficial effect was found to be more when the two compounds were fed in combination.

Antioxidant Potential of Stem Bark Extract of Shorea roxburghii against CCl 4 induced Liver Damage in Wistar Rats

The aim of the present study was to evaluate the antioxidant potential of the stem bark extract of Shorea roxburghii (S. roxburghii) on CCl4 induced liver toxicity. Stem bark extract of Shorea roxburghii (SRE) was prepared using ethyl acetate. Administration of S. roxburghii inhibited the elevation marker enzymes such as aminotransferase (AST), alkaline phosphatase (ALP) and alanine aminotransferase (ALT) and liver lipid peroxides in CCl4-treated Wistar rats. The natural antioxidant enzymes such catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) and lipid peroxidation were significantly improved after the treatment of S.roxburghii. In this study, the results indicate that S.roxburghii can inhibit lipid peroxidation and improve the activities of antioxidant enzymes. The quantitative analysis of biochemical parameters, marker enzymes, antioxidant activities and histological study suggest the administration of the stem bark extract of S.roxburghii protecting the liver from CCl4 toxicity.

Enhanced Hepatic Functions of Genetically Modified Mouse Hepatoma Cells by Spheroid Culture for Drug Toxicity Screening.

While hepatic cell lines are mainly used for in vitro drug induced toxicity studies, they exhibit limited functionalities. To overcome this, the authors have employed genetically engineered mouse hepatoma cells, Hepa/8F5, wherein expression of liver enriched transcription factors is induced by doxycycline leading to increased functionality. Further enhancement in functionality is achieved by spheroid culture in a previously developed 3D cell culture platform. Cells are seeded in presence of temperature-responsive poly(N-isopropylacrylamide) on poly(N-isopropylacrylamide--co-gelatin) cryogel scaffold based high throughput platform. Cells seeded in presence of poly(N-isopropylacrylamide) and induced with doxycycline exhibited highest functionalities. There is an increase of ≈26, 36, and 39% in albumin secretion, ammonia removal, and CYP3A4 activity, respectively. Morphological analysis showed arrest in cell proliferation and enlarged nucleus in presence of doxycyline and spheroid formation in presence of poly(N-isopropylacrylamide). Drug induced liver toxicity studies revealed that cells induced with doxycycline are resistive to tamoxifen but sensitive to acetaminophen whereas, cultures initiated in presence of poly(N-isopropylacrylamide) are resistive to both the drugs which is indicative of diffusional barrier of the spheroids. The authors conclude that Hepa/8F5 cells show enhanced functionality in cryogel based spheroid culture platform which can be successfully used for high throughput screening of hepatic toxicity in vitro.

Licochalcone E protects against carbon tetrachloride‑induced liver toxicity by activating peroxisome proliferator-activated receptor gamma.

The present study aimed to investigate the hepatoprotective role of Licochalcone E (LCE) and its mechanism of action in a mouse model of carbon tetrachloride (CCl4)-induced liver toxicity. Hepatotoxicity was induced in Kunming mice via an intraperitoneal injection (IP) of CCl4, 10 ml/kg body weight, diluted with corn oil at a 1:500 ratio. LCE was administered once a day for 7 days (IP) as pretreatment at a dose of 5 mg/kg/day. The levels of C-reactive protein (CRP) and tumor necrosis factor (TNF)-α were analyzed to determine the inflammation status. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed using ELISA assays. Liver ultrastructure was observed via optical microscopy. The mRNA and protein expression levels of peroxisome proliferator-activated receptor (PPAR)γ, and nuclear factor (NF)-κB were assayed using quantitative polymerase chain reaction and western blot analysis, respectively. Pretreatment with LCE decreased levels of ALT, AST, CRP and TNF-α, and NF-κB expression in the experimental hepatotoxicity mice model induced by CCl4. In addition, LCE increased the expression of PPARγ and normalized the hepatic histoarchitecture. However, the effects of LCE were reversed by cotreatment with the PPARγ inhibitor GW9662. The present study suggests that LCE may be used for the treatment of hepatotoxicity, and primarily exhibits its protective role through a PPARγ/NF-κB-mediated pathway.

HEPATOPROTECTIVE PROPERTIES OF SELECTED PLANTS AGAINST PARACETAMOL-INDUCED HEPATOTOXICITY IN MICE

The hepatoprotective properties of the ethanolic extracts of Punica granatum peels, Crataeva religiosa leaves and Jasminum polyanthrum leaves were evaluated for the paracetamol induced liver toxicity in mice. Antioxidant, cytotoxicity and phytochemical screening were carried out to assure the therapeutic efficacy of these plants. Oral administration of plant extracts for 7 days in mice significantly reduced the impact of paracetamol toxicity on the serum markers of liver damage, aspartate transaminase (AST), alanine transaminase (ALT) and total protein. The extract showed significant hepatoprotective effects as evidenced by decreased serum enzyme activities like ALT, AST and changed total protein. The phytochemical and antioxidant studies also supported that plant extract markedly reduced the toxicity due to presence of antioxidant phytochemicals. The result suggests that Jasminum polyanthrum is potent hepatoprotective agent against paracetamol induced hepatotoxicity in mice.Journal of Institute of Science and TechnologyVolume 22, Issue 1, July 2017, Page: 147-151

ANTIHEPATOTOXIC POTENTIALITY OF 2-3-6 TRIMETHYLOCT-6-ENAL AGAINST PARACETAMOL INDUCED LIVER DYSFUNCTION

Objective: We investigated the liver protective activity of 2-3-6 trimethyloct-6-enal from the methanol extract of Pajanelia longifolia (Willd.) K. Schuman. The liver protective activity of 2,3,6 trimethyloct-6-enal was evaluated against paracetamol (2 mg/kg body weight per orally) induced liver toxicity in swiss albino mice.Methods: Considering the Spectral data (IR spectrum, 1HNMR spectrum and 13C NMR spectrum) the predictable structure of 2,3,6 trimethyloct-6-enal was elucidated. To study the liver protective activity of the compound, Swiss albino mice of either sex were divided into six groups and treated for 5 d. Group I and II served as normal and toxic control, Group III were treated with Silymarin as a standard drug (50 mg/kg), and Group IV to VI was treated with 2-3-6 trimethyloct-6-enal at the dose of 50 mg/kg, 150 mg/kg and 250 mg/kg b.w. p. o. respectively. The liver protective activity of the compound was measured on biochemical parameters such as aspertate amino transferase (AST), alanine amino transferase (ALT), alkaline phosphatase (ALP), total bilirubin (TB), triglycerides (TGL), total cholesterol (TC) and protein. Further antioxidant activity of the compound was also measured on antioxidant enzymatic and non-enzymatic levels such as reduced glutathione (GSH), lipid peroxidation (LPO), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx)).Results: The study revealed that the compound has protective activity at the dose of 50, 150 and 250 mg/kg b.w. p. o. against paracetamol induced toxicity. In some biochemical parameters such as aspartate amino transferase and bilirubin, the compound has showed better result at a dose of 150 mg/kg compared to standard drug silymarin (value of aspartate amino transferase (compound) =71.10±0.12, (toxic) = 173.43±1.21, (silymarin) =79.86±0.02and total bilirubin (compound) = 1.04±0.11), (toxic) = 2.69±0.02, (silymarin) ==1.11±0.01. The findings were also confirmed by histopathological observations.Conclusion: 2,3,6 trimethyloct-6-enal from Pajanelia longifolia may be considered as a potent liver protective agent.

Discovery of phenylsulfonyl acetic acid derivatives with improved efficacy and safety as potent free fatty acid receptor 1 agonists for the treatment of type 2 diabetes

The free fatty acid receptor 1 (FFA1) has emerged as an attractive anti-diabetic target that mediates glucose-stimulated insulin secretion. Several FFA1 agonists have been reported, but many of them possessed somewhat high lipophilicity and/or molecular weight. Herein, we describe the identification of sulfone-carboxylic acid moiety with the multiple advantages of reducing lipophilicity, cytotoxicity and β-oxidation associated with compound 2. Further structure-activity relationship study based on the previleged scaffolds led to the discovery of 2-{(4-[(2'-chloro-[1,1'-biphenyl]-3-yl)methoxy]phenyl)sulfonyl}acetic acid (compound 20), which showed a better balance than compound 2 in terms of physicochemical properties, cytotoxicity profiles and pharmacokinetic properties. Subsequent in vivo studies demonstrated that compound 20 robustly improves the glucose tolerance both in normal and type 2 diabetic models without the risk of hypoglycemia. Compared to the high risk of TAK-875 induced liver toxicity, there was no significant adverse effects such as hepatic and renal toxicity were observed in the chronic toxicity studies of compound 20 even at the higher dose.

Exacerbation of N-nitrosodiethylamine Induced Hepatotoxicity and DNA Damage in Mice Exposed to Chronic Unpredictable Stress.

Psychological stress contributes to increased susceptibility to a number of diseases including cancer. The present study was designed to assess the effect of chronic unpredictable stress on N-nitrosodiethylamine induced liver toxicity in terms of in vivo antioxidant status and DNA damage in Swiss albino mice. The animals used in this study were randomized into different groups based on the treatment with N-nitrosodiethylamine or chronic unpredictable stress alone and post-stress administration of N-nitrosodiethylamine. The mice were sacrificed after 12 weeks of treatment, and the status of major enzymatic and non-enzymatic antioxidants, liver function markers, lipid peroxidation and the extent of DNA damage were determined in circulation and liver tissues of all the groups. The N-nitrosodiethylamine treated group showed significantly compromised levels of the antioxidant enzymes, lipid peroxidation, and the liver function markers with enhanced DNA damage as compared to chronic unpredictable stress or control groups. A similar but less typical pattern observed in the chronic unpredictable stress treated mice. All the measured biochemical parameters were significantly altered in the group treated with the combination of chronic unpredictable stress and N-nitrosodiethylamine when compared to controls, or chronic unpredictable stress alone and/or N-nitrosodiethylamine alone treated groups. Thus, exposure to continuous, unpredictable stress conditions even in general life may significantly enhance the hepatotoxic potential of N-nitrosodiethylamine through an increase in the oxidative stress and DNA damage.

Self microemulsifying formulation of Lagerstroemia speciosa against chemically induced hepatotoxicity

Self microemulsifying formulation is an approach used for enhancing the bioavailability of poorly soluble molecules due to their lipidic nature and small particle size. The objective of the present study was to evaluate the hepatoprotective activity of poorly soluble hydroxy- and polyhydroxy-organic phytomolecules rich Lagerstroemia speciosa leaves extract in modern formulation i.e. “Self microemulsifying System”.Different doses of SME (Self microemulsifying) formulation of L. speciosa leaves extract were evaluated for the hepatoprotective activity against carbon tetrachloride induced liver toxicity in rats. The parameters evaluated were (a) biochemical parameters like serum enzymes: aspartate aminotransferase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP) and total bilirubin (b) liver antioxidant parameters as estimation of Lipid peroxidation (LPO), catalase (CAT), Superoxide dismutase (SOD) activity and concentration of reduced glutathione (GSH). Oral administration of SME formulation provided the significant protection in marker enzyme of treated group at 100 mg/kg, p.o. as AST (P < 0.001), ALT (P < 0.001), ALP (P < 0.001) and total bilirubin (P < 0.001) comparable to the group treated with silymarin. Treatment with SME formulation at the doses of 100 mg/kg, p.o. significantly prevented the rise in levels of LPO significantly (P < 0.001). The GSH, SOD and CAT contents had significantly (P < 0.001) increased in SME formulation treated groups whereas carbon tetrachloride intoxicated group had shown significant decrease in these parameters compared to control group. Formulation at the dose 100 mg/kg, p.o. has shown maximum protection which was almost comparable to those of the normal control and standard. The histological observations further uphold the results for hepatoprotective activity.

Disruption of Estrogen Receptor Alpha in Rats Results in Faster Initiation of Compensatory Regeneration Despite Higher Liver Injury After Carbon Tetrachloride Treatment.

Estrogen receptor alpha (ESR1) is 1 of the 2 intracellular receptors for estrogen and is expressed by hepatocytes in the liver. The role of ESR1 in the regulation of toxicant-induced liver injury and compensatory regeneration is not completely clear. We investigated the role of ESR1 in liver regeneration after carbon tetrachloride (CCl4)-induced liver injury using wild type (WT) and ESR1 knockout (ESR1-KO) rats. Adult female WT and ESR1-KO rats were treated with 1 mL/kg CCl4 and euthanized over a time course of 0 to 48 hours. Liver injury measured by serum alanine amino transaminase, and histopathological analysis showed significantly higher liver injury in ESR1-KO as compared to WT rats. Hematoxylin and eosin staining revealed 2-fold higher necrosis and significant inflammatory cell infiltration in ESR1-KO rats. Chloracetate esterase staining revealed higher neutrophil infiltration in ESR1-KO rat livers. Interestingly, proliferating cell nuclear antigen immunohistochemistry showed that in spite of 2-fold higher liver injury, the ESR1-KO rats had equal liver regeneration as compared to WT rats. Western blot analysis of cyclin D1 and phosphorylated Rb, proteins involved in the initiation of the cell cycle, was significantly higher at all time points in ESR1-KO rats. Further analysis revealed faster activation of canonical Wnt/β-catenin and NF-κB signaling in ESR1-KO rats characterized by higher activated β-catenin and phosphorylated p65 at 12 hours after CCl4 treatment. Taken together, these data indicate that ESR1-mediated signaling inhibits liver regeneration by downregulation of Wnt signaling resulting in lower cyclin D1 activation after chemical-induced liver injury.

Improved MECP2 Gene Therapy Extends the Survival of MeCP2-Null Mice without Apparent Toxicity after Intracisternal Delivery

Intravenous administration of adeno-associated virus serotype 9 (AAV9)/hMECP2 has been shown to extend the lifespan of Mecp2−/y mice, but this delivery route induces liver toxicity in wild-type (WT) mice. To reduce peripheral transgene expression, we explored the safety and efficacy of AAV9/hMECP2 injected into the cisterna magna (ICM). AAV9/hMECP2 (1 × 1012 viral genomes [vg]; ICM) extended Mecp2−/y survival but aggravated hindlimb clasping and abnormal gait phenotypes. In WT mice, 1 × 1012 vg of AAV9/hMECP2 induced clasping and abnormal gait. A lower dose mitigated these adverse phenotypes but failed to extend survival of Mecp2−/y mice. Thus, ICM delivery of this vector is impractical as a treatment for Rett syndrome (RTT). To improve the safety of MeCP2 gene therapy, the gene expression cassette was modified to include more endogenous regulatory elements believed to modulate MeCP2 expression in vivo. In Mecp2−/y mice, ICM injection of the modified vector extended lifespan and was well tolerated by the liver but did not rescue RTT behavioral phenotypes. In WT mice, these same doses of the modified vector had no adverse effects on survival or neurological phenotypes. In summary, we identified limitations of the original vector and demonstrated that an improved vector design extends Mecp2−/y survival, without apparent toxicity.

Study of the Antifibrotic Effect of Olmesartan on the Carbon Tetrachloride- Induced liver Toxicity in Rats

Study of the Antifibrotic Effect of Olmesartan on the Carbon Tetrachloride- Induced liver Toxicity in Rats

Sa1805 - Novel Immunomodulatory Role of Aspartate and Nmda Receptor in Acetaminophen Induced Liver Toxicity and Acute Pancreatitis

Sa1805 - Novel Immunomodulatory Role of Aspartate and Nmda Receptor in Acetaminophen Induced Liver Toxicity and Acute Pancreatitis

Hepatoprotective effect of sitagliptin against methotrexate induced liver toxicity.

Sitagliptin is selective dipeptidyl peptidase-4 inhibitor (DPP4-I), used clinically as a new oral anti-diabetic agent. This study explored the underlying mechanisms of the hepatoprotective role of sitagliptin pretreatment against methotrexate (MTX) induced hepatotoxicity in mice. Forty mice were divided into four groups (10 mice each); control, MTX, and two sitagliptin groups (pretreated with sitagliptin 10 and 20 mg/kg/day, respectively) for five consecutive days prior to MTX injection. Results showed that MTX induced marked hepatic injury in the form of cloudy swelling, hydropic degeneration, apoptosis and focal necrosis in all hepatic zones. Biochemical analysis revealed significant increase in the serum transaminases, alkaline phosphatase and lactate dehydrogenase in MTX group. Oxidative stress and depressed antioxidant system of the hepatic tissues were evident in MTX group. MTX down-regulated nuclear factor erythroid 2-related factor 2 (Nrf2) expression and reduced its binding capacity. Additionally, MTX increased the activation and the expression of nuclear factor kappa-B (NF-κB) and downstream inflammatory mediators. MTX induced the activation of inducible nitric oxide synthase (iNOS) and increased nitrite/nitrate level. Finally, hepatic cellular apoptosis was clearly obvious in MTX-intoxicated animals using TUNEL staining. Also, there was increase in the immunoexpression of pro-apoptotic protein Bax, increase in Bax and caspase-3 levels and decrease in the level of anti-apoptotic Bcl2 in liver. On the other hand, sitagliptin pretreatment significantly ameliorated all of the above mentioned biochemical, histopathological, immunohistochemical changes induced by MTX. These results provide new evidences that the hepatoprotective effect of sitagliptin is possibly mediated through modulation of Nrf2 and NF-κB signaling pathways with subsequent suppression of inflammatory and apoptotic processes.

Amikacin-induced liver toxicity: correlations between biochemical indexes and ultrastructural features in an experimental model

In previous studies, aminoglycosides (AG) as gentamicin (G), dibekacin (D), tobramycin (T), netilmicin (N) and Sysomicin (S) were proved to induce ultrastructural alterations in the liver of experimental animals. The aim of this studies is to investigate the effect of amikacin (AK) on rabbit liver which is commonly used in infections resistant to other AG; this was done studying both the common blood parameters and ultrastructural changes. The study was accomplished in 24 New-Zealand rabbits, twelve received 20 mg/kg AK every 12 hours for 2 weeks. Thereafter the animals were anesthesized and liver slices were obtained for transmission electron microscopy. As results obvious signs of primary and secondary microcholestasis associated to mitochondrial cristae detachment and phospholipid aggregations were noted; this last finding was less marked when compared to previous studies employing other AG. In the AK treated group, blood tests showed a significant increased in only Blood Urea Nitrogen (BUN) and an insignificant rise in AST levels. Our findings are consistent with an AK induced liver toxicity albeit less evident with respect to the other AG.

HLA-B*57 Allele Is Associated with Concomitant Anti-tuberculosis and Antiretroviral Drugs Induced Liver Toxicity in Ethiopians.

Drug-induced liver injury (DILI) is a known adverse effect of both anti-tuberculosis (anti-TB) and antiretroviral (ARV) drugs. Recent studies highlight the implications of genetic predispositions to DILI. We performed a case-control study to identify Human Leukocyte Antigen-B (HLA-B) variant alleles associated with anti-TB and ARV co-treatment induced liver toxicity in Ethiopian TB and HIV co-infected patients. A total of 495 newly diagnosed TB and HIV co-infected patients were enrolled and received rifampicin based anti-TB and efavirenz based ARV therapy. Change in liver enzyme level from baseline was monitored 1st, 2nd, 4th, 8th, 12th, and 24th weeks after treatment initiation to identify patients who developed DILI (cases) and those who did not (treatment tolerants). Genomic DNA from 46 cases and 46 sex and age matched treatment tolerants were genotyped for HLA-B variant alleles using Olerup SSP®HLA-B DNA Typing Kits. The proportion of HLA-B*57 allele carriers in DILI cases (37.0%), particularly in those who developed cholestatic type of DILI (44.8%) was significantly higher compared with those who tolerated the treatment (2.2%). The HLA-B*57 allele frequency was significantly higher in cases (25%) than treatment tolerants (1.1%). In a multivariate logistic analysis, the proportion of patients carrying HLA-B*57 (P = 0.002) and HLA-B*14 (P = 0.014) alleles were significantly higher in DILI cases compared with treatment tolerants. HLA-B*57 was significantly associated with cholestatic (P = 0.001) and mixed (P = 0.017) types of liver toxicity, and mild-to-moderate severity (P = 0.001). Of all HLA-B*57 alleles detected, HLA-B*57:03 accounted 58.3% and HLA-B*57:02 accounted 41.7%. HLA-B*57:01 was not detected. The variant allele frequencies of HLA-B*57:03 (15.2 vs. 0%) and HLA-B*57:02 (9.8 vs. 1.1%) were significantly higher in the DILI cases than treatment tolerants (P < 0.03). We conclude that HLA-B*57 alleles (B*57:03 and B*57:02) confer susceptibility to the development of anti-TB and ARV drugs co-treatment induced liver toxicity, which is mainly of cholestatic type. The possible association of HLA-B*14 with anti-TB and ARV drugs co-treatment induced liver toxicity requires further investigations.

Hepatotoxicity of Eupatorium adenophorum extracts and the identification of major hepatotoxic components

Eupatorium adenophorum can induce liver toxicity in animals. For the safe utilisation of the weed, the hepatotoxic components need to be discovered. In this study, in vitro hepatotoxicity of different extracts from E. adenophorum were determined on human hepatocyte cell line L02 and hepatocellular carcinoma cell line HepG2. The results showed that water extracts of E. adenophorum exhibited no hepatotoxicity in vitro while high concentrations of the organic solvent extracts had obvious hepatotoxicity. Sesquiterpenes may contribute to the toxicity based on the comparison of composition analysis. Three cadinene sesquiterpenes were purified and identified as 9-oxo-10,11-dehydroageraphorone, 10Hα-9-oxo-ageraphorone and 10Hβ-9-oxo-ageraphorone. In vitro hepatotoxic effects of these components were investigated, the IC50 of the three compounds were 122.53, 87.52, and 108.80 μM in L02 cells and 151.92, 104.48, and 138.08 μM in HepG2 cells by Cell Counting Kit-8 (CCK-8) assay. The three components were confirmed to be, at least partial, hepatotoxic components.