Hepatic Malondialdehyde Content Research Articles

Effects of fishmeal replacement by Tenebrio molitor meal on growth performance, antioxidant enzyme activities and disease resistance of the juvenile pearl gentian grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀ )

A 50-day feeding trial was conducted for juvenile pearl gentian grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀) to determine the optimal amount of Tenebrio molitor meal (TM) that could replace fishmeal (FM) in feeds without reducing growth performance, antioxidant enzyme activities and resistance. Six isonitrogenous and isolipidic diets were formulated with TM replacing 0% (T0), 6.25% (T6), 12.5% (T12), 18.75% (T18), 25% (T25), 31.25% (T31) FM respectively. Each diet was randomly divided into triplicate groups with 30 juveniles (initial weight 6.60 ± 0.00 g). After feeding trail, fish were challenged by Vibrio harveyi for 7 days. The results showed that each treatment had better survival rate between 96.67% and 100%. Fish fed with control and T6 diet showed lower final weight and weight gain rate as compared to other diets. The feed conversion ratio of fish fed on T12 and T18 diet had no significant difference compared to control diet. As compared to control diet, other diets had no significant differences on specific growth rate and temperature growth coefficient. Fish fed with T6 diet had highest liver glutathione reductase activities. No significant differences were observed in liver superoxide dismutase activities in fish fed with T6, T12, T25, T31 and control diet. Fish fed on T18 diet had lowest hepatic malondialdehyde contents and a highest relative per cent survival. According to a broken-line model, the recommended substituted ratio of TM for FM was estimated to be 12.3% for grouper obtaining maximum WGR, corresponding to additive amount of TM is 4.92%.

Alpha lipoic acid protects against chlorpyrifos-induced toxicity in Wistar rats via modulating the apoptotic pathway

The chronic exposure to chlorpyrifos (CPF) pesticide induces several human disorders including hepatotoxicity. Alpha-lipoic acid (ALA) is a natural antioxidant compound found in plants and animals. The present study aimed to investigate the possible protective effect of ALA against CPF-induced hepatotoxicity and the possible underlying molecular mechanism. Thirty-two male Wistar rats were divided into: Normal rats received only vehicle; ALA group received ALA (10 mg/kg, i.p.); CPF group received CPF (18 mg/kg, s.c.) and CPF-ALA group received CPF (18 mg/kg, s.c.) once daily for 14 days. The present results demonstrated that administration of ALA significantly improved liver functions (p < 0.05) and limited the histopathological lesions induced by CPF in liver tissues. Furthermore, ALA decreased hepatic malondialdehyde contents while increased the glutathione peroxidase, catalase, superoxide dismutase and acetylcholinesterase activities. Interestingly, ALA showed significant antiapoptotic effects through downregulation of Bax and Caspase-3 expression levels. In conclusion, ALA possess protective effects against CPF-induced liver injury through attenuation of apoptosis and oxidative stress.

Nigella sativa oil modulates the therapeutic efficacy of mesenchymal stem cells against liver injury in irradiated rats

Stem cell transplantation is a novel strategy for regenerative medicine in liver disease. This study was conducted to explore the modulatory effect of Nigella sativa oil (NSO) on the therapeutic potential of mesenchymal stem cells (MSCs) against irradiation-induced liver damage in rats. Liver damage was induced by a total body exposure to a single dose of 7Gy. NSO (2mg/kg/day) was then given orally for 4 consecutive weeks starting 24h after irradiation with or without a single intravenous MSCs administration, then rats were sacrificed four weeks after exposure to γ radiation. Data revealed that irradiation elevated aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in serum, increased hepatic malondialdehyde (MDA) content and reduced hepatic superoxide dismutase (SOD) activity. Furthermore, it caused elevation in pro-inflammatory mediators such as interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) associated with reduction in anti-inflammatory cytokine interleukin-10 (IL-10) and it increased fibrogenic marker transforming growth factor-β (TGF-β) in liver tissues. It was observed that combined NSO/MSCs therapy provided more beneficial tissue repair comparable to MSCs alone as demonstrated by modulating the tested parameters. Finally, these results were confirmed by histopathological examination. In conclusion, dual therapy with NSO and MSCs could serve as a promising approach for alleviating radiation-induced liver injury in patients with radiotherapy.

Caspase-Mediated Anti-Apoptotic Effect of Ginsenoside Rg5, a Main Rare Ginsenoside, on Acetaminophen-Induced Hepatotoxicity in Mice.

Frequent overdose of acetaminophen (APAP) is one of the most common and important incentives of acute hepatotoxicity. Prior to this work, our research group confirmed that black ginseng (Panax ginseng, BG) showed powerful protective effects on APAP-induced ALI. However, it is not clear which kind of individual ginsenoside from BG plays such a liver protection effect. The objective of the current investigation was to evaluate whether ginsenoside Rg5 (G-Rg5) protected against APAP-induced hepatotoxicity and the involved action mechanisms. Mice were administrated with G-Rg5 at two dosages of 10 or 20 mg/kg for 7 consecutive days. After the last treatment, all of the animals that received a single intraperitoneal injection of APAP (250 mg/kg) showed severe liver toxicity after 24 h, and the liver protection effects of G-Rg5 were examined. The results clearly indicated that pretreatment with G-Rg5 remarkably inhibited the production of serum tumor necrosis factor (TNF-α) and interleukin-1β (IL-1β) compared with the APAP group. Meanwhile, G-Rg5 decreased the hepatic malondialdehyde (MDA) content, the protein expression levels of 4-hydroxynonenal (4-HNE) and cytochrome P450 2E1 (CYP2E1) in the liver tissues. G-Rg5 decreased APAP caused the hepatic overexpression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). Furthermore, analysis of immunohistochemistry and Western blotting also indicated that G-Rg5 pretreatment inhibited activation of apoptotic pathways mainly via increasing the expression of Bcl-2 protein, decreasing the expression of Bax protein, proliferating cell nuclear antigen (PCNA), cytochrome c, caspase-3, caspase-8, and caspase-9. Liver histopathological observation provided further evidence that pretreatment with G-Rg5 could significantly inhibit hepatocyte necrosis, inflammatory cell infiltration, and apoptosis caused by APAP. In conclusion, the present study clearly demonstrates that G-Rg5 exerts a liver protection effect against APAP-induced acute hepatotoxicity mainly via a caspase-mediated anti-apoptotic effect.

Sildenafil protects against bile duct ligation induced hepatic fibrosis in rats: Potential role for silent information regulator 1 (SIRT1)

Hepatic fibrosis is a potential health problem that may end with life-threatening cirrhosis and primary liver cancer. Recent studies point out to the protective effects of silent information regulator1 (SIRT1), against different models of organs fibrosis. This work aimed to investigate the possible protective effect of sildenafil (SIRT1 activator) against hepatic fibrosis induced by bile duct ligation (BDL). Firstly, three different doses of sildenafil (5, 10, 20mg/kg/day) were investigated; to detect the most protective one against BDL induced liver dysfunction and hepatic fibrosis. The most protective dose is then used; to study its effect on BDL induced SIRT1 downregulation, imbalance of oxidant/antioxidant status, increased inflammatory cytokines and fibrosis. Sildenafil (20mg/kg/day) was the most protective one, it caused upregulation of SIRT1, reduction of hepatic malondialdehyde (MDA) content, increase in expression of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenease (HO)-1, reduced glutathione (GSH) content and superoxide dismutase (SOD) activity. Hepatic content of tumor necrosis factor-α (TNF-α) and nuclear factor κB (NFκB) expression & content displayed significant reductions with sildenafil treatment, Furthermore, sildenafil caused marked reductions of transforming growth factor (TGF)-β content, expression of plasminogen activator inhibitor-1 (PAI-1), matrix metalloproteinase-9 (MMP-9), tissue inhibitor of metalloproteinase-1 (TIMP-1), α-smooth muscle actin (α-SMA), fibronectin, collagen I (α1) and hydroxyproline content. However, sildenafil protective effects were significantly reduced by co-administration of EX527 (SIRT1 inhibitor). Our work showed, for the first time that, sildenafil has promising protective effects against BDL induced liver dysfunction and hepatic fibrosis. These effects may be, in part, mediated by up regulation of SIRT1.

The effect of DL-, L-isomers and DL-hydroxy analog administered at 2 levels as dietary sources of methionine on the metabolic and antioxidant parameters and growth performance of turkeys

A hypothesis was verified that dietary methionine (Met) improves the growth and antioxidant status of turkeys, and that its effects depend on dietary inclusion levels and sources. A total of 816 female Hybrid Converter turkeys was fed wheat-soybean meal-based diets supplemented with 3 sources of Met: DL-, L-isomers and DL-hydroxy analog (DLM, LM, and MHA, respectively). In 4 4-week periods (from one to 16 wk of age), dietary Met content corresponded to NRC (1994) recommendations or was increased by approximately 50% (in one to 8 wk by 44 to 46% and in 9 to 16 wk by 55 to 56% vs. the NRC guidelines) to match the recommendations of some breeding companies. Increased Met content resulted in higher final body weights of turkeys (P = 0.002), an improved feed conversion ratio (P = 0.049), increased total glutathione concentration and ferric reducing ability of plasma (FRAP) values, and decreased malondialdehyde (MDA) concentration (all P < 0.001) in the blood plasma of turkeys. In comparison with DLM, LM and MHA contributed to an increase in plasma glutathione concentration (P = 0.001), a decrease in plasma triacylglycerol (P = 0.003) and uric acid (P = 0.001) concentrations, and a decrease in liver MDA (P = 0.001) levels. A decrease in plasma MDA (vs. DLM) and lipid peroxides (LOOH) (vs. DLM and LM) concentrations as well as a decrease in plasma superoxide dismutase (SOD) activity (vs. DLM and LM) also were noted in the MHA treatment (P = 0.016, P = 0.001 and P = 0.011, respectively). In conclusion, the results of the study indicate that the antioxidant status of turkeys could be affected by dietary Met levels and sources. The dietary Met content increased by 50% relative to NRC recommendations, improved the growth performance of turkeys, and strengthened their antioxidant defense system. In comparison with DLM, LM and MHA could be considered positive nutritional factors as manifested by a beneficial decrease in plasma and hepatic MDA concentrations as well as an increase in plasma glutathione levels, and the effect of MHA was more pronounced.

The enantioselective toxicity and oxidative stress of beta-cypermethrin on zebrafish

Although the toxicity of beta-cypermethrin (beta-CYP) to aquatic organisms has become a significant concern in recent years, its enantioselective effects on non-target organisms is poorly understood. To investigate the enantioselective toxicity of beta-CYP on zebrafish, adult zebrafish were exposed to a series of isometric concentrations of four beta-CYP enantiomers and the beta-CYP racemate for 96 h. In addition, the activities of four antioxidant enzymes and the malondialdehyde (MDA) content in zebrafish liver and brain were tested after 15 and 30 days beta-CYP enantiomers and racemate exposure under environmentally relevant dosages (0.01 and 0.1 μg/L). According to the acute toxicity results, the 1R-cis-αS and 1R-trans-αS enantiomers were more lethal than 1S-cis-αR and 1S-trans-αR. At 0.1 μg/L, the 1R-cis-αS and 1R-trans-αS enantiomers, and the beta-CYP racemate could significantly induce a hepatic MDA content at 30 days post exposure (dpe), while only 1R-cis-αS caused brain lipid peroxidation. An apparent regulation of antioxidase levels was observed in zebrafish liver and brain after exposure to the 1R-cis-αS and 1R-trans-αS enantiomers, and the beta-CYP racemate. In contrast, no significant oxidative stress was observed in zebrafish exposed to 1S-cis-αR and 1S-trans-αR enantiomers under the test concentrations. This work demonstrated the occurrence of enantioselectivity in toxicity and oxidative stress of beta-CYP to adult zebrafish, which should be considered in environmental risk assessments.

Regulation of growth, tissue fatty acid composition, biochemical parameters and lipid related genes expression by different dietary lipid sources in juvenile black seabream, Acanthopagrus schlegelii

The effects of different dietary lipid sources on physiological characteristics and lipid metabolism relevant genes expression were assessed in juvenile black seabream Acanthopagrus schlegelii. 720 black seabream juveniles (initial weight was 5.4±0.0g) were randomly allocated in 18 floating net cages corresponding to triplicate cages of the six dietary treatments, and the 8-week feeding trail was conducted. Five isonitrogenus and isolipidic diets were formulated to contain lipid sources: coconut oil (CO), perilla oil (PO), sunflower oil (SO), arachidonic acid (ARA) and eicosapentaenoic acid combined with docosahexaenoic acid (EPA+DHA), and fish oil (FO) was set as a control group. The results showed that weight gain (WG), specific growth rate (SGR) and feed efficiency (FE) in fish fed diets containing FO and EPA+DHA were significantly higher than CO. In serum, the highest activity of glutathione peroxidase (GSH-PX) and glutathione (GSH) content were recorded in fish fed EPA+DHA. The significantly highest content of malondialdehyde (MDA) was observed in fish fed the EPA+DHA and FO diets. The maximum hepatic content of MDA and the minimum activity of superoxide dismutase (SOD) were all found in fish fed the diet containing EPA+DHA. High contents of cholesterol (CHOL) and triacylglycerol (TAG) were found in fish fed SO, whereas glucose (GLU) content in fish fed EPA+DHA was significantly higher than in the other treatments. Fish fed FO had the highest hemoglobin (HGB) among treatments. The anabolic pathway relevant genes: accα, fas and srebp-1 were up-regulated by dietary SO and PO. Whereas, the catabolic pathway relevant genes: lpl, cpt1a, atgl, hsl and pparα were down-regulated by dietary CO. Furthermore, the long chain polyunsaturated fatty acid (LC-PUFA) biosynthesis relevant genes elovl5 and fads2 were significantly up-regulated by dietary SO and PO. The present study focused on comparing the effects of different dietary lipid sources on biochemical characteristics and lipid metabolism relevant genes expression in black seabream. Overall, it demonstrated that physiological-biochemical characteristics as well as lipid metabolism relevant genes expression were significantly regulated by different dietary lipid sources. Moreover, dietary PO and SO regulated some physiological and biochemical indicators. These findings could contribute to optimize aquatic feeds when considering selection of optimal lipid sources or their optimal proportion in the future.

Saponins (Ginsenosides) from the Leaves of Panax quinquefolius Ameliorated Acetaminophen-Induced Hepatotoxicity in Mice.

Acetaminophen (APAP) overdose is one of the most common inducements of drug-induced liver injury (DILI) in the world. The main purpose of this paper was to investigate the liver protection activity of saponins (ginsenosides) from the leaves of Panax quinquefolius (PQS) against APAP-induced hepatotoxicity, and the involved mechanisms were demonstrated for the first time. Mice were pretreated with PQS (150 and 300 mg/kg) by oral gavage for 7 days before being treated with 250 mg/kg APAP. Severe liver injury was exerted at 24 h post-APAP, and hepatotoxicity was assessed. Our results showed that pretreatment with PQS significantly decreased the serum alanine aminotransferase (ALT), aspartate transaminase (AST), tumor necrosis factor (TNF-α), and interleukin-1β (IL-1β) levels in a dose-dependent manner as compared to the APAP administration. Meanwhile, compared with that in the APAP group, PQS decreased hepatic malondialdehyde (MDA) contents and 4-hydroxynonenal (4-HNE) expression and restored reduced glutathione (GSH) content and superoxide dismutase (SOD) activity in livers of mice. PQS inhibited the overexpression of pro-inflammatory factors cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) in the liver tissues. Furthermore, Western blotting analysis revealed that PQS pretreatment inhibited the activation of apoptotic signaling pathways via increase of Bcl-2 and decrease of Bax and caspase-3 protein expression levels. Liver histopathological observation provided further evidence that PQS pretreatment significantly inhibited APAP-induced hepatocyte necrosis, inflammatory cell infiltration, and congestion. Biological indicators of nitrative stress such as 3-nitrotyrosine (3-NT) were inhibited after PQS pretreatment, compared to the APAP group. The present study clearly demonstrates that PQS exerts a protective effect against APAP-induced hepatic injury because of its antioxidant, anti-apoptotic, and anti-inflammatory activities. The findings from the present investigation show that PQS might be a promising candidate treatment agent against drug-induced ALI.

Strain differences in arsenic-induced oxidative lesion via arsenic biomethylation between C57BL/6J and 129X1/SvJ mice

Arsenic is a common environmental and occupational toxicant with dramatic species differences in its susceptibility and metabolism. Mouse strain variability may provide a better understanding of the arsenic pathological profile but is largely unknown. Here we investigated oxidative lesion induced by acute arsenic exposure in the two frequently used mouse strains C57BL/6J and 129X1/SvJ in classical gene targeting technique. A dose of 5 mg/kg body weight arsenic led to a significant alteration of blood glutathione towards oxidized redox potential and increased hepatic malondialdehyde content in C57BL/6J mice, but not in 129X1/SvJ mice. Hepatic antioxidant enzymes were induced by arsenic in transcription in both strains and many were higher in C57BL/6J than 129X1/SvJ mice. Arsenic profiles in the liver, blood and urine and transcription of genes encoding enzymes involved in arsenic biomethylation all indicate a higher arsenic methylation capacity, which contributes to a faster hepatic arsenic excretion, in 129X1/SvJ mice than C57BL/6J mice. Taken together, C57BL/6J mice are more susceptible to oxidative hepatic injury compared with 129X1/SvJ mice after acute arsenic exposure, which is closely associated with arsenic methylation pattern of the two strains.

Dietary effects of soy and citrus flavonoid on antioxidation and microbial quality of meat in broilers.

The current study investigated dietary effects of soy genistein (SG) and citrus hesperidin (CH) on microbial spoilage and antioxidant quality of chicken meat. Broilers were divided to six treatment groups: control (C), SG and CH, supplemented with 5mg of SG and 20mg of CH per kg of feed, respectively, and SGCH1, SGCH2 and SGCH3, supplemented with 5, 10 or 20mg of SG+CH (1:4) per kg of feed respectively. Both SG and CH reduced (p<.01) the microbial load of spoilage organisms in breast meat at 0 and 15days of refrigeration. Hepatic and muscle total antioxidant and superoxide dismutase activities, and crude protein contents were generally improved (p<.05), whereas hepatic malondialdehyde concentration and muscle fat was decreased (p<.01) by the dietary treatments. In conclusion, soy genistein and CH supplementation could improve meat composition, antioxidant and microbial quality of broilers meat.

Effects of pentachlorophenol on the quail (Coturnix japonica) liver detoxification pathway

Pentachlorophenol (PCP), an extensively used pesticide and biocide, is of critical environmental concern due to its toxicity and recalcitrance to degradation. In this study, the effect of PCP on induction of transcription factors, cytochrome P450 (CYP450) genes, and the antioxidative enzyme system were investigated in the quail liver. A total of 60 (4- to 6-week-old) male quails (Coturnix japonica) were administered 0, 0.05, 0.5, and 5 mg/kg/d PCP orally for 42 d. Following exposure, both absolute and relative liver weights were significantly lower than those of the control. Using gas chromatography-mass spectrometry, PCP accumulation was, from highest to lowest, kidney > liver > muscle for all exposure groups. The expressions of CYP1A5, CYP1B1, CYP2C18, nuclear translocator 1 (ARNT1), and aryl hydrocarbon receptor 1 (AHR1) were induced after PCP treatment, and increases were found in the activities of hepatic superoxide dismutase (SOD) and catalase (CAT), and the content of hepatic malondialdehyde (MDA). In addition, exposure to PCP induced an increase in liver 8-hydroxydeoxyguanosine (8-OHdG) and significantly elevated ethoxyresorufin O-deethylase (EROD), methoxyresorufin O-demethylase (MROD), and 7-ethoxycoumarin-O-deethylase (ECOD) activity, but decreased that of glutathione peroxidase (GSH-Px), benzyloxyresorufin O-debenzylase (BROD), pentoxyresorufin O-depentylase (PROD), and erythromycin N-demethylase (END). No significant responses were observed for benzyloxy-trifluoromethyl-coumarin (BFC). The protein level of liver nuclear factor κB (NF-κB) was higher, whereas that of nuclear factor E2-related factor 2 (Nrf2) was lower for exposed quail. These results suggest that PCP affects quail oxidative stress by modulating CYP450 enzymes and nuclear transcription factors.

Formononetin protects against acetaminophen-induced hepatotoxicity through enhanced NRF2 activity.

To examine the effects of formononetin (FMN) on Acetaminophen (APAP)-induced liver injury in vitro and in vivo. Human non-tumor hepatic cells LO2 were pretreated with either vehicle or FMN (20, 40 μM), for 6 h, followed by incubation with or without APAP (10 mM) for 24 h. In an in vivo assay, male BALB/c mice were randomly divided into four groups: (1) control group; (2) APAP group; (3) APAP + FMN (50 mg/Kg); (4) APAP + FMN (100 mg/Kg). The mice in the control and APAP groups were pre-treated with vehicle; the other two groups were pretreated daily with FMN (50, 100 mg/Kg) orally for 7 consecutive days. After the final treatment, acute liver injury was induced in all groups, except the control group, by intraperitoneal (i.p.) injection of 300 mg/Kg APAP. In LO2 cells, APAP exposure decreased the cell viability and glutathione (GSH) content, which were both greatly restored by FMN pretreatment. Overdose of APAP increased hepatic malondialdehyde (MDA) content, serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activity in experimental mice. Supplementation with 100 mg/Kg FMN significantly reduced APAP-induced elevated levels of MDA (1.97 ± 0.27 vs 0.55 ± 0.14 nmol/mg protein, p < 0.001), ALT (955.80 ± 209.40 vs 46.90 ± 20.40 IU/L, p < 0.001) and AST (1533.80 ± 244.80 vs 56.70 ± 28.80 IU/L, p < 0.001), and hepatic GSH level (5.54 ± 0.93 vs 8.91 ± 1.11 μmol/mg protein, p < 0.001) was significantly increased. These results were further validated by histopathology and TdT-mediated biotin-dUTP nick-endlabeling (TUNEL) staining, pretreatment with 100 mg/Kg FMN significant decreased APAP-induced hepatocellular damage and cell apoptosis (36.55 ± 3.82 vs 2.58 ± 1.80%, p < 0.001). Concomitantly, FMN stimulated the expression of Nrf2 and antioxidant gene expression in the presence of APAP. These data provide an experimental basis for the use of FMN in the treatment of patients with APAP-induced hepatotoxicity.

Virgin coconut oil supplementation attenuates acute chemotherapy hepatotoxicity induced by anticancer drug methotrexate via inhibition of oxidative stress in rats.

The emerging health benefit of virgin coconut oil (VCO) has been associated with its potent natural antioxidants; however, the antioxidant and hepatoprotective effect of VCO against methotrexate-induced liver damage and oxidative stress remains unexplored. The study explored the antioxidant and hepatoprotective effects of VCO against oxidative stress and liver damage induced by anticancer drug methotrexate (MTX) in rats. Liver damage was induced in Wistar rats pretreated with dietary supplementation of VCO (5% and 15%) by intraperitoneal administration of MTX (20mg/kg bw) on day 10 only. After 12days of treatment, assays for serum liver biomarkers (aminotransferases), alkaline phosphatase, albumin and total protein as well as hepatic content of malondialdehyde, reduced glutathione and antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase) were carried out. Liver was used to examine histopathological changes. MTX administration induced significant increase in serum liver enzymes along with marked decrease in albumin and total protein compared to control group. Hepatic activities of antioxidant enzymes were significantly decreased, while malondialdehyde increased significantly. Treatment with VCO supplemented diet prior to MTX administration attenuated MTX-induced liver injury and oxidative stress evidenced by significant improvements in serum liver markers, hepatic antioxidant enzymes and malondialdehyde comparable to control group. Histopathological alterations were prevented and correlated well with the biochemical indices. The study suggests antioxidant and hepatoprotective effects of VCO supplementation against hepatotoxicity and oxidative damage via improving antioxidant defense system in rats. Our findings may have beneficial application in the management of hepatotoxicity associated with MTX cancer chemotherapy.

Protective effects of petroleum ether extracts of Herpetospermum caudigerum against α-naphthylisothiocyanate-induced acute cholestasis of rats

Ethnopharmacological relevanceThe ripe seeds of Herpetospermum caudigerum have been used in Tibetan folk medicine for treatment of bile or liver diseases including jaundice, hepatitis, intumescences or inflammation. Previously reports suggested that the seed oil and some lignans from H. caudigerum exhibited protective effects against carbon tetrachloride (CCl4)-induced hepatic damage in rats, which may be related to their free radical scavenging effect. However, the protective effect of H. caudigerum against cholestasis is still not revealed. The aim of the present study was to investigate the pharmacological effect and the chemical constituents of the petroleum ether extract (PEE) derived from H. caudigerum against α-naphthylisothiocyanate (ANIT)-induced acute cholestasis in rats. Materials and methodsMale cholestatic Sprague-Dawley (SD) rats induced by ANIT (60mg/kg) were orally administered with PEE (350, 700 and 1400mg/kg). Levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-Glutamyl transpeptidase (γ-GTP), total bilirubin (TBIL), direct bilirubin (DBIL) and total bile acid (TBA), as well as bile flow, and histopathological assay were evaluated. Hepatic malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione S-transferase (GST), and nitric monoxide (NO) in liver were measured to explore the possible protective mechanisms. Phytochemical analysis of PEE was performed by gas chromatography-mass spectrometer (GC-MS). ResultsPEE have exhibited significant and dose-dependent protective effect on ANIT-induced liver injury by reduce the increases in serum levels of ALT, AST, ALP, γ-GTP, TBIL, DBIL and TBA, restore the bile flow in cholestatic rats, and reduce the severity of the pathological tissue damage induced by ANIT. Hepatic MDA, MPO and NO contents in liver tissue were reduced, while SOD and GST activities were elevated in liver tissue. 49 compounds were detected and 39 of them were identified by GC-MS analysis, in which long-chain fatty acids were the main constituents. ConclusionsPEE exhibited a dose-dependently protective effect on ANIT-induced liver injury in cholestatic rats with the potential mechanism of attenuated oxidative stress in the liver tissue, and the possible active compounds were long-chain fatty acids.

Protective Effects of Ginsenosides on 17[Formula: see text]-Ethynyelstradiol-Induced Intrahepatic Cholestasis via Anti-Oxidative and Anti-Inflammatory Mechanisms in Rats.

The present study was designed to assess the effects and potential mechanisms of ginsenosides on 17[Formula: see text]-ethynyelstradiol (EE)-induced intrahepatic cholestasis (IC). Ginsenoside at doses of 30, 100, 300[Formula: see text]mg/kg body weight was intragastrically (i.g.) given to rats for 5 days to examine the effect on EE-induced IC. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and total bile acid (TBA) were measured. Hepatic malondialdehyde (MDA) content and superoxide dismutase (SOD) activity were determined. Protein expression of proinflammatory cytokines TNF-[Formula: see text], IL-6 and IL-1[Formula: see text] was analyzed by immunohistochemistry and Western blot. Results indicated that ginsenosides remarkably prevented EE-induced increase in the serum levels of AST, ALT, ALP and TBA. Moreover, the elevation of hepatic MDA content induced by EE was significantly reduced, while hepatic SOD activities were significantly increased when treated with ginsenosides. Histopathology of the liver tissue showed that pathological injuries were relieved after treatment with ginsenosides. In addition, treatment with ginsenosides could significantly downregulate the protein expression of TNF-[Formula: see text], IL-6 and IL-1[Formula: see text] compared with EE group. These findings indicate that ginsenosides exert the hepatoprotective effect on EE-induced intrahepatic cholestasis in rats, and this protection might be attributed to the attenuation of oxidative stress and inflammation.

Active form of vitamin D ameliorates non-alcoholic fatty liver disease by alleviating oxidative stress in a high-fat diet rat model.

The purpose of this study was to determine whether treatment using the active form of vitamin D (1,25(OH)2D3) could protect against high-fat diet (HFD)-induced non-alcoholic fatty liver disease (NAFLD) in rats and ameliorate oxidative stress. Male Sprague-Dawley rats were divided into three groups and treated with standard chow, HFD, or HFD plus intraperitoneal injection of 1,25(OH)2D3 (5 μg/kg body weight, twice per week), respectively, for 16 weeks. Serum lipid profiles, hepatic function, intrahepatic lipid, and calcium levels were determined. Hepatic histology was examined using hematoxylin/eosin, Masson's trichrome, and Oil Red O staining. Oxidative stress was assessed by measuring hepatic malondialdehyde (MDA) and F2α-isoprostane content. Expression of nuclear factor-erythroid-2-related factor 2 (Nrf2) and downstream target genes was analyzed using quantitative RT-PCR. 1,25(OH)2D3 treatment improved the serum lipid profile, reduced intrahepatic lipid levels, and attenuated hepatic steatosis and inflammation in HFD rats. Furthermore, MDA and F2α-isoprostane levels in liver tissue were reduced by 1,25(OH)2D3 administration. Although 1,25(OH)2D3 did not regulate the expression of Nrf2 mRNA, it did induce Nrf2 nuclear translocation. The expression of Nrf2 target genes, including Gclc, Nqo1, Sod2, and Cat, was up-regulated by 1,25(OH)2D3. We conclude that 1,25(OH)2D3 protects against HFD-induced NAFLD by attenuating oxidative stress, inducing NRF2 nuclear translocation, and up-regulating the expression of genes encoding antioxidant enzymes.

Elevated Apoptosis in the Liver of Dairy Cows with Ketosis

Background/Aims: Dairy cows with ketosis are characterized by oxidative stress and hepatic damage. The aim of this study was to investigate hepatic oxidative stress and the apoptotic status of ketotic cows, as well as the underlying apoptosis pathway. Methods: The blood aspartate aminotransferase (AST), alanine aminotransferase (ALT), glutamate dehydrogenase (GLDH) and gamma-glutamyl transferase (GGT) activities and the haptoglobin (HP), serum amyloid A (SAA) and serum apoptotic cytokeratin 18 neo-epitope M30 (CK18 M30) concentrations were determined by commercially available kits and ELISA kits, respectively. Liver histology, TUNEL and Oil red O staining were performed in liver tissue samples. TG contents were measured using an enzymatic kit; Caspase 3 assays were carried out using the Caspase 3 activity assay kit; oxidation and antioxidant markers were measured using biochemical kits; apoptosis pathway were determined by qRT-PCR and western blot. Results: Ketotic cows displayed hepatic fat accumulation. The hepatic malondialdehyde (MDA) content was significantly increased, but the activities of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were markedly decreased in ketotic cows compared with control cows, indicating that ketotic cows displayed severe oxidative stress. Significantly higher serum levels of the hepatic damage markers AST, ALT, GGT and GLDH were observed in ketotic cows than in control cows. The blood concentration of the apoptotic marker CK18 M30 and the number of TUNEL-positive cells in the liver of ketotic cows were 1.19- and 2.61-fold, respectively, higher than the values observed in control cows. Besides, Caspase 3 activity was significantly increased in the liver of ketosis cows. Importantly, the levels of phosphorylated c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38MAPK) were significantly increased but the level of phosphorylated extracellular signal-regulated kinase1/2 (ERK1/2) was markedly decreased, which further promoted tumor protein 53 (p53) expression and inhibited nuclear factor E2-related factor 2 (Nrf2) expression. The apoptosis-related molecules p21, MDM2, Caspase 3, Caspase 9 and Bax were expressed at significantly higher levels in ketotic cows than in healthy cows, whereas the anti-apoptosis molecule Bcl-2 was expressed at significantly lower levels. Conclusions: Based on these results, ketotic cows display severe hepatic oxidative stress. The hepatic MAPK-p53-Nrf2 apoptotic pathway is over induced and partially mediated apoptotic damage in the liver.

N-acetylcysteine attenuates intrauterine growth retardation-induced hepatic damage in suckling piglets by improving glutathione synthesis and cellular homeostasis.

The objective of the present study was to test the hypothesis that N-acetylcysteine (NAC) may play beneficial roles against intrauterine growth retardation (IUGR)-induced hepatic damage in suckling piglets. Fourteen IUGR and seven normal birth weight (NBW) neonatal male piglets were selected. Piglets were weaned at 7days of postnatal age and fed the control formula milk (NBW-CON and IUGR-CON groups) or the control formula milk supplemented with 1.2g/kg NAC (IUGR-NAC group) for 14days (n=7). The plasma and liver samples were analyzed for the parameters related to hepatic damage, redox status, apoptosis, and autophagy. Compared with the NBW-CON group, IUGR-CON group exhibited increased activities of plasma aminotransferases, increased numbers of apoptotic hepatocytes, as well as higher concentrations of protein carbonyl, malondialdehyde (MDA), microtubule-associated protein 1 light chain 3 beta, and phospholipid-conjugated form (MAP1LC3B-II), along with a decrease in the content of reduced glutathione (GSH). NAC treatment increased GSH content and GSH-to-oxidized GSH ratio in the liver of IUGR-NAC group, most likely owing to the improved activities of γ-glutamine-cysteine ligase, γ-glutamine-cysteine synthetase, and glutathione reductase. The hepatic protein carbonyl and MDA contents were decreased in the IUGR-NAC group compared with the IUGR-CON group. In addition, NAC-treated piglets had an increased content of B cell lymphoma/leukemia 2 protein, whereas a decreased expression level of MAP1LC3B-II in the liver. NAC may have beneficial effects in improving GSH synthesis and cellular homeostasis in the liver of IUGR suckling piglets.

Ameliorative Effect of Gallic Acid on Methotrexate-Induced Hepatotoxicity and Nephrotoxicity in Rat.

We investigated the protective effect of gallic acid (GA) against methotrexate (MTX)-induced hepatotoxicity and nephrotoxicity. Male Wistar rats were randomized into five groups (n = 6/group): I, control; II, MTX-treated for seven days; III, pre-treated with GA for seven days, followed by MTX for seven days; IV, co-treated with MTX and GA for seven days and V, GA for seven days. MTX caused a significant increase (P<0.05) in plasma biomarkers of nephrotoxicity (urea, creatinine) and hepatotoxicity (Bilirubin, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase, gamma glutamyl transferase) when compared with control. Furthermore, MTX caused a significant decrease in the activities of hepatic enzymic antioxidants (superoxide dismutase, catalase, glutathione S-transferase) and nonenzymic antioxidants (Vitamin C and glutathione), followed by a significant increase in hepatic malondialdehyde content. However, pre-treatment and co-treatment with gallic acid ameliorated the MTX-induced biochemical changes observed. Taken together, GA protected against MTX-induced hepatotoxicity and nephrotoxicity in rats, by reducing the impact of oxidative damage to tissues.