Decreased Serum AST Research Articles

Deletion of protein kinase C θ attenuates hepatic ischemia/reperfusion injury and further elucidates its mechanism in pathophysiology.

Hepatic ischemia-reperfusion (HIR) is a severe process in pathophysiology that occurs clinically in hepatectomy, and hepatic transplantations. The present study aimed to investigate the effect of PKC θ deletion against HIR injury and elucidate its mechanism in pathophysiology. HIR injury was induced in wild-type and PKC θ deletion mice treated with or without heme. The ALT and AST levels were determined to evaluate liver function. HIR injury was observed via histological examination. Oxidative stress and inflammatory response markers, and their signaling pathways were detected. The study found that PKC θ knockout decreased serum AST and ALT levels when compared to the WT mice. Furthermore, heme treatment significantly reduced the ALT and AST levels of the PKC θ deletion mice compared with the untreated PKC θ deletion mice. PKC θ deletion markedly elevated superoxide dismutase activity in the liver tissue, reduced malondialdehyde content in the tissue, and the serum TNF-α and IL-6 levels compared with the WT mice. Heme treatment was observed to elevate the activity of SOD and reduced MDA content and serum of TNF-α and IL 6 in the PKC θ deletion animals. Meanwhile, heme treatment increased HO-1 and Nrf 2 protein expression, and reduced the levels of TLR4, phosphorylated NF-κB, and IKB-α. These findings suggested that PKC θ deletion ameliorates HIR, and heme treatment further improves HIR, which is related to regulation of PKC θ deletion on Nrf 2/HO-1 and TLR4/NF-κB/IKB α pathway.

Piezo1 alleviates acetaminophen-induced acute liver injury by activating Nrf2 and reducing mitochondrial reactive oxygen species

Acetaminophen (APAP) overdose is the most common cause for acute liver failure (ALF) in the developed countries, with limited treatment options. Piezo1 is a mechanosensitive cation channel. We found that APAP caused upregulation of Piezo1 in both an APAP-induced acute liver injury (ALI) animal model and a mouse hepatocyte cell line AML12. Activation of Piezo1 by its activator Yoda1 reduced APAP-induced hepatotoxicity and ROS level. Mechanistically, activation of Piezo1 led to accumulation of the antioxidant regulator Nrf2 and upregulation of its target genes Nqo1 and Gsta1, while knockdown of Piezo1 downregulated this pathway. Finally, injection of Yoda1 decreased serum AST and ALT levels, reduced cell death and rescued liver injury in the APAP-induced ALI mouse model. Our findings suggested a previously undiscovered protective role of Piezo1 in APAP-induced ALI, which might shed light on a new therapeutic target for this disease.

Zinc-glutathione in Chinese Baijiu prevents alcohol-associated liver injury

Zinc depletion is associated with alcohol-associated liver injury. We tested the hypothesis that increasing zinc availability along with alcohol consumption prevents alcohol-associated liver injury. Zinc-glutathione (ZnGSH) was synthesized and directly added to Chinese Baijiu. Mice were administered a single gastric dose of 6 g/kg ethanol in Chinese Baijiu with or without ZnGSH. ZnGSH in Chinese Baijiu did not change the likeness of the drinkers but significantly reduced the recovery time from drunkenness along with elimination of high-dose mortality. ZnGSH in Chinese Baijiu decreased serum AST and ALT, suppressed steatosis and necrosis, and increased zinc and GSH concentrations in the liver. It also increased alcohol dehydrogenase and aldehyde dehydrogenase in the liver, stomach, and intestine and reduced acetaldehyde in the liver. Thus, ZnGSH in Chinese Baijiu prevents alcohol-associated liver injury by increasing alcohol metabolism timely with alcohol consumption, providing an alternative approach to the management of alcohol-associated drinking.

Leptadenia hastata Leaf Extract ameliorates oxidative stress and serum biochemical parameters in Streptozotocin-Induced diabetes in Wistar rats.

: Diabetes Mellitus is a major health problem characterized by hyperglycemia and disturbances in metabolism and implicated in causing oxidative stress. Treatment includes administration of oral hypoglycaemic agents with lifestyle modifications, these offer glycemic control, however, present limitations about availability, affordability and side effects. Traditional anti-diabetic plants are becoming popular in management of diabetes mellitus. This study was carried out to determine the efficacy of Leptadenaia hastata in treatment of diabetes. Diabetes mellitus was induced in using a single injection of streptozotocin (50mg kg- 1 i.p.). The rats were divided into four groups of 5 rats each. Groups 3-6 received olive oil, 100mg kg- 1 extract, 200mg.kg- 1 extract and insulin (6IU kg- 1), respectively. 10 non-diabetic rats were grouped into two group receiving olive oil and 200mg kg- 1 extract for 28 days. All groups were sacrificed by injecting with ketamine hydrochloride, blood was collected by cardiac puncture and centrifuged. The serum was analyzed for biochemical parameters. The liver was removed and homogenized with the supernatant of the resultant homogenate collected and used for analysis of oxidative stress enzymes. The extract significantly decreased serum AST (p < 0.05), ALP (p < 0.001), ALT (p < 0.05), TG (p < 0.01), TC (p < 0.001), creatinine (p < 0.001). It had no effect on SOD and CAT levels but it significantly increased (p < 0.001) GSH levels and reduced (p < 0.05) MDA level. The n-hexane extract of Leptadenia hastata significantly decreased the levels of hepatic and renal serum biomarkers proving that it was beneficial in ameliorating diabetic related complications. The extract significantly increased GSH levels and reduced MDA level. The online version contains supplementary material available at 10.1007/s40200-022-01017-z.

Liraglutide attenuates hepatic iron levels and ferroptosis in db/db mice

ABSTRACT Liver pathological changes are as high as 21%-78% in diabetic patients, and treatment options are lacking. Liraglutide is a glucagon-like peptide-1 (GLP-1) receptor that is widely used in the clinic and is approved to treat obesity and diabetes. However, the specific protection mechanism needs to be clarified. In the present study, db/db mice were used to simulate Type 2 diabetes mellitus (T2DM), and they were intraperitoneally injected daily with liraglutide (200 μg/kg/d) for 5 weeks. Hepatic function, pathologic changes, oxidative stress, iron levels, and ferroptosis were evaluated. First, liraglutide decreased serum AST and ALT levels, and suppressed liver fibrosis in db/db mice. Second, liraglutide inhibited the ROS production by upregulating SOD, GSH-PX, and GSH activity as well as by downregulating MDA, 4-HNE, and NOX4 expression in db/db mice. Furthermore, liraglutide attenuated iron deposition by decreasing TfR1 expression and increasing FPN1 expression. At the same time, liraglutide decreased ferroptosis by elevating the expression of SLC7A11 and the Nrf2/HO-1/GPX4 signaling pathway in the livers of db/db mice. In addition, liraglutide decreased the high level of labile iron pools (LIPs) and intracellular lipid ROS induced by high glucose in vitro. Therefore, we speculated that liraglutide played a crucial role in reducing iron accumulation, oxidative damage and ferroptosis in db/db mice.

Hepatoprotective effect of anemoside B4 against sepsis-induced acute liver injury through modulating the mTOR/p70S6K-mediated autophagy

Sepsis triggers liver dysfunction with high morbidity and mortality. Here, we elucidated the effect of anemoside B4 on sepsis in cecal ligation and puncture (CLP)-induced mouse model and LPS-induced primary hepatocytes. Following CLP surgery, septic mice were intraperitoneally injected with anemoside B4 (50 or 100 mg/kg). Anemoside B4 improved septic mouse survival rate, decreased serum AST and ALT levels and attenuated liver histopathologic damages. Western blot analysis showed that anemoside B4 elevated the expression of Beclin-1, LC3II/LC3I, Atg3, Atg5, and Atg7, and reduced p62, suggesting the restoration of autophagy flux in liver. More autophagic vesicles were observed in liver after anemoside B4 treatment using transmission electron microscopy. Using ELISA and commercial enzyme kits, we found that anemoside B4 decreased serum TNF-α, IL-6, and IL-1β levels and increased CAT, SOD and GSH activities. TUNEL staining and western blot revealed that anemoside B4 suppressed cell apoptosis, along with decreased Bax, leaved caspase-3, cleaved PARP, but increased Bcl-2. Consistent with in vivo findings, anemoside B4 inhibited apoptosis, inflammatory response, and oxidative stress and enhanced autophagy in LPS-induced primary hepatocytes. Importantly, these cellular processes were possibly mediated by mTOR/p70S6K signaling, as reflected by the offset of 3-MA in the immunosuppression of anemoside B4.

Therapeutic Potential And Hepatoprotective Activity Of Proanthocyanidins And In Combination With Clopidogrel Against Bimolecular Disturbances Related To Non Alcoholic Fatty Liver Disease Induced In Rats

Nonalcoholic fatty liver disease (NAFLD) is a complex disease caused by a number of different pathogenic processes as a result of the systemic interaction between the liver and several other organs. Grape seeds proanthocyanidins extract (GSPE) potency in the liver is associated with improvement of the hepatic enzyme activities as their powerful antioxidant property results from its ability to directly scavenge free radicals and/or chelate metals. 32 male albino rats were assigned into 4 equal groups of 8 rats: 1. Normal control group (G1) of rats provided with standard normal diet for 12 weeks, 2. NAFL group (G2) of rats received high fat diet (HFD)-feeding for 6 weeks for induction of NAFL, followed by normal feeding for other 6 weeks, 3. GSPE treated group (G3) of the HFD-induced NAFL, followed by administration of GSPE for 6 weeks else, 4. GSPE and antiplatletes drug (Clopidogrel) co-treated group (G4), of the NAFL rats, followed by administration of GSPE and clopidogrel co-treatment for another 6 weeks. The results revealed that GSPE alone or with clopidogrel treatment significantly decreased serum AST, ALT and γGGT (G3, 4) that were significantly increased in HFD-NAFL rats (G2), and showed significant upregulation of IL1β, PPARα, TGFβ1 and TIMP1 in livers of NAFL rats (G2), that was significantly downregulated following administration of GSPE alone (G3), or with clopidogrel (G4). Subsequantly, treatment with GSPE alone or with antiplatelet drug can improve the metabolic disruptions associated with the induction of NAFL by improving the liver biomarkers and inflammatory mediator's expressions.

Chemical and biological effects of some mixtures of plant oils and Thymus vulgaris on liver diseases

Liver is the vital organ with a wide range of functions that can influence other body organs. Dietary components are essential for the healthy or diseased liver. Selected food plants can provide nutritional and medicinal support for liver diseases. Herbal and oil medicinal products are increasingly being used and many of them have shown promising potential for the treatment of liver diseases. The aim of this study is to investigate the protective role of thyme, olive oil and flaxseed oil on carbon tetrachloride CCl4 induced changes in liver enzymes of albino rats. Sixty adult male Albino rats weighing about 130±5 g were taken and divided into twelve groups, each with five rats. The first group is the control (-) and fed on normal diet for 10 weeks. The second group received subcutaneous injection with CCL4 in paraffin oil (50% v/v 2 ml/kg) twice per week for 2 weeks to induce chronic damage in the liver tissue and fed on normal diet (control +). Another experimental groups (n = 5/group) were fed a Commercial diet with different doses from thyme, flaxseed oil and olive oil for 10 weeks. There were a significant increase in the activities of serum ALT and AST in rats of the positive control group (+) as compared to negative control group (-) (P ≤ 0.05).While the protected groups with high doses of thyme (4,6,8,10,12) and olive oil and flaxseed oil (3,5,7,9 ,11 ) recorded a significant decreased serum AST and ALT enzyme compared to the low doses groups. Recommendations and Conclusion: High doses of thyme, olive oil, and flaxseed oil could ameliorate carbon tetrachloride (CCl4)-induced liver injury in rats, which. Suggesting that diet rich in flaxseed oil, olive oil and thyme might be a promising approach for the prevention of liver diseases.

Estrogen protects against liver damage in sepsis through inhibiting oxidative stress mediated activation of pyroptosis signaling pathway.

Sepsis was characterized by systemic inflammatory response and multisystem organ dysfunction, refering to the activation of inflammatory and oxidative stress pathways. Estrogen has been shown to have anti-inflammatory and antioxidant effects as well as extensive organ protective role. However, whether estrogen alleviates sepsis-induced liver injury and the mechanisms involved remain unknown. Septic mice were constructed by intraperitoneal injection lipopolysaccharide, and the effect of estrogen on liver injury was investigated. Furthermore, the roles of NLRP3 inhibitor MCC950 and mitochondrial ROS specific scavenger Mito-tempo, on the liver injury were explored in septic mice. Female septic mice exhibited liver damage with increased serum AST and ALT level as well as the existence of extensive necrosis, and which was more serious in male septic mice. Moreover, Ovariectomy (OVX) aggravated sepsis-induced liver damage and activation of pyroptosis signaling pathway, which was alleviated by estrogen as evidenced by decreased serum AST, ALT level and number of infiltrating inflammatory cell, as well as protein expression related to pyroptosis. OVX aggravated mitochondrial dysfunction and liver injury in septic mice was also partly reversed by Mito-tempo and MCC950. These results demonstrated that estrogen protected against sepsis-induced liver damage through alteration of mitochondrial function and activation of inflammatory-mediated pyroptosis signaling pathway.

Dietary administration of the probiotic Shewanella putrefaciens to experimentally wounded gilthead seabream (Sparus aurata L.) facilitates the skin wound healing

The effect of the probiotic Shewanella putrefaciens Pdp11 (SpPdp11) was studied on the skin healing of experimentally wounded gilthead seabream (Sparus aurata L.). Two replicates (n = 12) of fish were fed CON diet or SP diet for 30 days. Half of the fish were sampled while the others were injured and sampled 7 days post-wounding. Results by image analysis of wound areas showed that SpPdp11 inclusion facilitated wound closure. Compared with the CON group, fish in SP group sampled 7 days post-wounding had a significantly decreased serum AST and increased ALB/GLOB ratio. Furthermore, protease and peroxidase activities were significantly increased in skin mucus from fish in SP group sampled 7 days post-wounding, compared with those fed CON diet. Additionally, SP diet up-regulated the gene expression of antioxidant enzymes, anti-inflammatory cytokines, and re-epithelialization related genes in the fish skin. Furthermore, significant decreases in pro-inflammatory cytokines expression were detected in fish from SP group, respect to control ones. Overall, SpPdp11 inclusion facilitated the wound healing and the re-epithelialization of the damaged skin, alleviated the inflammatory response in the wound area through intensifying the antioxidant system, and enhancing the neo-vascularization and the synthesis of matrix proteins in the skin wound sites of fish.

Beet root (Beta vulgaris) protects lipopolysaccharide and alcohol-induced liver damage in rat.

The purpose of this study was to investigate the protective effects of beet root (Beta vulgaris var. rubra) in lipopolysaccharide (LPS) and alcohol induced liver damage. Beta vulgaris ethanol extract (BVEE) showed good antioxidant activity in the contents of polyphenol and flavonoid compounds, and the electron-donating ability and ABTS+ radical scavenging activity. As for anti-inflammatory effect in RAW 264.7 cells, inhibition rate of nitric oxide production was increased in dose dependent manner. In hepatotoxicity model induced by LPS and alcohol in rat, BVEE significantly decreased serum AST, ALT and γ-GTP concentrations in a dose-dependent manner. The histopathological changes after H&E staining showed that fat accumulation and inflammatory cell infiltration were decreased by BVEE. The collagen fibers around the central lobule observed by Masson's trichrome staining were also decreased by BVEE. In addition, as for the immunohistochemical staining and Transmission electron microscopy, BVEE improved morphological characteristics of damaged liver lesion. The increased mRNA expressions of NF-κB, MAPK1, MAPK3, CYP2E1, and α-SMA were significantly decreased in BVEE treated group. These results indicated that BVEE would have protective effects in hepatotoxicity by altering various indicators related to the liver damage induced by LPS or alcohol.

The impact of Lactobacillus acidophilus on hepatic and colonic fibrosis induced by ethephon in a rat model.

Objective(s):The study is aimed to elucidate the impact of antioxidant, anti-inflammatory and antifibrosis properties of Lactobacillus acidophilus (L. acidophilus) on liver and colon in ethephon treated rats through measuring Pro- inflammatory cytokines, oxidative stress index, lysosomal cathepsin-D enzyme activity and fibrosis markers.Materials and Methods:Rats divided into three groups; Group 1: distilled water control, Group 2: rats at day 16 from experiment beginning were orally received ethephon 50 mg/ kg BW in distilled water once daily for 60 days. Group 3: rats were orally received L. acidophilus enriched diet 1% (w/w) for 15 days as prophylactic, then received both L. acidophilus enriched diet 1% (w/w) and ethephon 50 mg/kg BW for 60 days. Results:Ethephon exerts hepatic and colonic oxidative stress, inflammatory response and fibrosis through NF-κB activation. On contrary, L. acidophilus supplementation evokes hepatoprotective properties as revealed by decreased serum AST, ALT, γ -GT and increased IGF-1. L. acidophilus exerts antioxidant and anti-inflammatory properties as indicated by decreased TOS, OSI, TNF-α, IL-1β, cathepsins D activity, NF-κB expression and increased TAC, lysosomal membrane stability. L. acidophilus shows antifibrotic activity as demonstrated by down-regulation of TGF-β1, α-SMA, collagen expression. Conclusion: L. acidophilus possess antioxidant, anti -inflammatory and antifibrotic activity through inhibition of NF-kB.

Gallic acid produces hepatoprotection by modulating EGFR expression and phosphorylation in induced preneoplastic liver foci in rats

The purpose of this study was to analyze the role of gallic acid as liver protector and identify its role in the regulation of EGFR expression and phosphorylation in induced preneoplastic liver lesions in rats. Male Wistar rats were randomly divided into four groups. (1) Control; (2) animals receiving gallic acid (AG) 50 mg/kg v.o. for 8 weeks; (3) animals with preneoplasia (P) induced by a single dose of diethylnitrosamine 200 mg/kg i.p. (DEN) and two weeks after a single dose of carbon tetrachloride 2 mL/kg i.p. (CCl4); and (4) animals with preneoplasia treated with GA during 8 weeks. In order to evaluate GA hepatoprotection on preneoplastic lesions, we performed histological examination of liver tissue using H&E staining as well as an immunohistochemical analysis for PCNA. To evaluate the effect of GA on EGFR expression and phosphorylation, we performed an immunohistochemical and western blot analysis. The results indicated that GA significantly decreased EGFR expression and pY1068 EGFR phosphorylation in animals with preneoplastic lesions. GA significantly decreased PCNA expression in animals with preneoplastic lesions, suggesting it may work as an antiproliferative agent. Additionally, GA improved the architecture and organization of liver tissue and significantly decreased serum AST, ALT and FA, which are indicators of hepatocellular damage. By histopathological and immunohistochemical analysis we demonstrated an improvement in liver morphology, a reduction of preneoplastic liver foci and a reduction of cell proliferation, as well as an improvement on liver functionality. In conclusion, GA produces hepatoprotection by modulating EGFR expression and phosphorylation in preneoplastic lesions.

Protective Effects the Akt Activator SC79 in Hepatic Ischemia-Reperfusion Injury.

BackgroundSC79 has been reported to protect against experimental ischemia-elicited neuronal death and brain injury and to protect myocardiocytes from hypoxia/reoxygenation (H/R) injury. Here, we investigated the effects of SC79 in primary hepatocytes in vitro and in rat liver in vivo following hypoxia-reoxygenation (H/R) and hepatic I/R injury.Material/MethodsThe livers of Sprague-Dawley rats were subjected to 45 min of ischemia followed by 2–24 h of reperfusion. The primary hepatocytes were subjected to hypoxia for 6 h and for 2–24 h. The hepatocytes cells or the hepatic I/R injury model livers were treated with SC79 or/and LY294002 at different times and concentrations. The serum ALT, AST, histologic examination, cellular viability, and cell apoptosis were assessed. The levels of phospho-Akt, Bad, Bim, Bax, Bcl-2, and Bcl-XL were determined by Western blot analysis.ResultsSC79 improved viability and inhibited apoptosis in hepatocytes following H/R. SC79 decreased serum AST and ALT, markedly improved pathology, and decreased cell apoptosis in livers following I/R. In addition, SC79 promoted the expression of phospho-Akt, Bcl-2, and Bcl-XL, and decreased the expression of Bid, Bax, and Bim. PI3K inhibitor (LY294002) pre-treatment completely abolished the above-mentioned effects of SC79.ConclusionsThe protective role of SC79 against H/R of hepatocytes or hepatic I/R injury is related to activation of phosphorylation of Akt, resulting in the decrease of pro-apoptotic protein of Bim, Bax, and Bad, and increase of the anti-apoptotic protein Bcl-2 and Bcl-xL induced by cell H/R and hepatic I/R injury.

A Controlled Fermented Samjunghwan Herbal Formula Ameliorates Non-alcoholic Hepatosteatosis in HepG2 Cells and OLETF Rats.

Hepatosteatosis (HS), a clinical feature of fatty liver with the excessive intracellular accumulation of triglyceride in hepatocytes, is manifested by perturbation of the maintenance of liver lipid homeostasis. Samjunghwan (SJH) is an herbal formula used mostly in Korean traditional medicine that is effective against a number of metabolic diseases, including obesity. Herbal drugs, enriched with numerous bioactive substances, possess health-protective benefits. Meanwhile, fermented herbal products enriched with probiotics are known to improve metabolic processes. Additionally, current lines of evidence indicate that probiotics-derived metabolites, termed as postbiotics, produce the same beneficial effects as their precursors. Herein, the anti-HS effects of 5-weeks naturally fermented SJH (FSJH) was investigated with FSJH-mixed chow diet in vivo using Otsuka Long-Evans Tokushima Fatty (OLETF) and Long-Evans Tokushima Otsuka (LETO) rats as animal models of HS and controls, respectively. In parallel, the anti-HS effects of postbiotic-metabolites of three bacterial strains [Lactobacillus brevis (LBB), Lactococcus lactis (LCL) and Lactobacillus plantarum (LBP)] isolated from FSJH were also evaluated in vitro using the FFAs-induced HepG2 cells. Feeding OLETF rats with FSJH-diet effectively reduced body, liver, and visceral adipose tissue (VAT) weights, produced marked hypolipidemic effects on serum and hepatic lipid parameters, decreased serum AST and ALT levels, and upregulated the HMGCOR, SREBP, and ACC, and downregulated the AMPK and LDLR gene expressions levels. Additionally, exposure of FFAs-induced HepG2 cells to postbiotic metabolic media (PMM) of bacterial strains also produced marked hypolipidemic effects on intracellular lipid contents and significantly unregulated the HMGCOR, SREBP, and ACC, and downregulated the AMPK and LDLR genes expressions levels. Overall, our results indicate that FSJH enriched with fermented metabolites could be an effective anti-HS formulation.

Hepatocyte miRNA‐21‐Deficiency Promotes Hepatic Lipid Accumulation but Ameliorates Alcohol‐Induced Liver Injury by Targeting DUSP16

BackgroundAccumulation of lipid droplets and inflammatory cell infiltration in the liver is the pathophysiological hallmark of alcoholic liver disease (ALD). The cell‐type specific role of miR‐21 in ALD remains unknown due to the lack of cell‐type specific knockout mice. The current study aims at elucidating the pathophysiological role of miR‐21 in ALD using our newly generated hepatocyte specific miR‐21 knockout mice.MethodsWe generated a new mouse model for hepatocyte specific deletion of miR‐21 (Hep‐KO). To induce alcoholic steatosis, Hep‐KO and their wild‐type (WT) littermates were fed with isovolumetric/isocaloric control liquid diet or Lieber‐DeCarli ethanol liquid diet at different concentrations (1% to 6%) for 4 weeks followed by a single binge. Markers of steatosis, liver injury and inflammation were assessed. Lipidomics and metabolomics were conducted. Other methods: liver histology, 3′UTR luciferase reporter, qPCR, Western blots, Oil O red staining.ResultsLiver miR‐21 RNA levels were highly induced in WT mice fed the ethanol diet compared with mice fed the control diet. Moreover, miR‐21 levels were markedly upregulated in human patients with alcoholic cirrhosis vs normal controls, demonstrating their roles in the pathogenesis of ALD. Hep‐KO mice fed the ethanol diet had increased hepatic triglyceride (TG) but decreased plasma TG levels, which was associated with increased lipogenesis (FASN, SCD1 and SREBP protein expression) but decreased VLDL secretion (MTTP). Lipidomics analysis revealed increased TG but decreased phosphatidyl cholines (PC) and phosphatidyl ethanolamines (PE) lipid species in Hep‐KO vs WT mice. Interestingly, inflammation was attenuated in Hep‐KO, as evident by the decreased serum AST and ALT levels, and reduced expression of key hepatic pro‐inflammatory (Il‐6, Ccr2, MIP1a, LyG6) genes. At the cellular level, MAPKs, AMPK and PPARs signaling pathways were assessed to determine the key regulatory role of miR‐21 in steatosis and inflammation, which showed that the activation of P38 and JNK were suppressed in Hep‐KO. Using TargetScan, microrna and miRanda, we predicted a conserved miR‐21 seed match region in mouse and human DUSP16 3′UTRs; a gene known as a phosphatase of P38 and JNK. Transfection of miR‐21 mimics reduced the DUSP16 protein in HepG2 and Hepa1 cells. In addition, the mouse DUSP16 3′UTR containing the miR‐21 seed region was cloned into a luciferase reporter and miR‐21 mimic inhibited DUSP16 3′UTR activity in a dose‐dependent fashion, which was relieved when the miR‐21 seed region was mutated. Furthermore, DUSP16 protein expression was down‐regulated in patients with alcoholic liver cirrhosis, where miR21, P‐P38 and P‐JNK levels were elevated.ConclusionHepatocyte miR‐21‐deficiency promotes alcoholic steatosis, but ameliorates liver injury and inflammation through the miR‐21/DUSP16/P38 pathway. Our work provides novel insights into the understanding of the molecular mechanism of non‐coding RNAs as key regulators in alcoholic liver disease.Support or Funding InformationGrant support: NIH R01DK104656, R01DK080440, R01ES025909, R21AA022482, R21AA024935, VA Merit Award 1I01BX002634.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Acanthoic acid protectsagainst ethanol-induced liver injury: Possible role of AMPK activation and IRAK4 inhibition

The aim of this study was to investigate the effects of acanthoic acid (AA) on the regulation of inflammatory response, lipid accumulation, and fibrosis via AMPK- IRAK4 signaling against chronic alcohol consumption in mice. Ethanol-induced liver injury was induced in male mice by Lieber-DeCarli diet for 28d. And mice in AA groups were gavaged with AA (20 or 40mg/kg) for 28d. AA treatment significantly decreased serum AST and TG, hepatic TG levels, serum ethanol and LPS levels compared with chronic ethanol administration. AA ameliorated histological changes, lipid droplets, hepatic fibrosis, and inflammation induced by ethanol. AA significantly increased the expressions of p-LKB1, p-AMPK, and SIRT1 caused by chronic ethanol administration, and attenuated the increasing protein expressions of IRAK1 and IRAK4.siRNA against AMPKα1 blocked AMPKα1 and increased IRAK4 protein expressions, compared with control-siRNA-transfected group, while AA treatment significantly decreased IRAK4 expressions compared with AMPKα1-siRNA-transfected group. AMPK-siRNA also blocked the decreased effect of AA on inflammatory factors. AA decreased over-expression of IRAK4 and inflammation under ethanol plus LPS challenge. AA recruited LKB1-AMPK phosphorylation and activated SIRT1 to regulate alcoholic liver injury, especially, inhibited IRAK1/4 signaling pathway to regulate lipid metabolism, hepatic fibrosis and inflammation caused by alcohol consumption.

Anti-Diabetic and Hepato-Renal Protective Effects of Ziyuglycoside II Methyl Ester in Type 2 Diabetic Mice.

Type 2 diabetes is a metabolic disorder caused by abnormal carbohydrate metabolism, and closely associated with abnormal lipid metabolism and hepato-renal dysfunction. This study investigated the anti-diabetic and hepato-renal protective properties of ziyuglycoside I (ZG01) derivative on type 2 diabetes. ZG01 was isolated from roots of Sanguisorba officinalis and chemically modified by deglycosylation and esterification to obtained ziyuglycoside II methyl ester (ZG02-ME). Here, we showed that ZG02-ME has stronger anti-diabetic activity than the original compound (ZG01) through decreasing blood glucose, glycated hemoglobin (HbA1c), and insulin levels in a mouse model of type 2 diabetes (db/db mice). We further found that ZG02-ME treatment effectively ameliorated serum insulin, leptin and C-peptide levels, which are key metabolic hormones, in db/db mice. In addition, we showed that elevated basal blood lipid levels were decreased by ZG02-ME treatment in db/db mice. Furthermore, treatment of ZG02-ME significantly decreased serum AST, ALT, BUN, creatinine, and liver lipid peroxidation in db/db mice. These results demonstrated that compared to ZG01, chemically modified ZG02-ME possess improved anti-diabetic properties, and has hepato-renal protective activities in type 2 diabetes.

Hepatoprotective effects of glycyrrhizin and omega-3 fatty acids on Nuclear Factor-kappa B pathway in thioacetamide-induced fibrosis in rats

Nuclear Factor kappa B (NF-κB) is a key transcriptional regulator that plays important roles in the pathogenesis of hepatic inflammation and fibrosis in chronic liver diseases. NF-κB activation leads to production of pro-inflammatory and fibrogenic cytokines. Glycyrrhizin (GL) is reported to suppress liver fibrosis and cirrhosis. Omega-3 fatty acids (ω-3) play an anti-inflammatory role and they are reported to decrease hepatic injury in Thioacetamide (TAA) fibrotic model. We investigated the effects of GL and ω-3 on liver inflammation and fibrosis in rats and clarified the effects of these natural compounds on NF-κB level. 50 male Wistar rats randomized to 5 groups: Control group and 4 groups received TAA 200 mg/kg i.p. twice weekly for 8 weeks: TAA group, GL group (received GL 25 mg/kg daily by oral tube), ω-3 group (received ω-3150 mg/kg daily by oral tube), (GL+ω-3) group (received similar combined doses of both natural compounds). GL and ω-3 alone or in combination protected the liver from TAA hepatotoxic effects as they significantly decreased serum AST activity and serum total bilirubin level, they also significantly increased serum albumin and total protein levels. The hepatoprotective effects of GL and ω-3 were confirmed by the histopathological analysis as they significantly reduced the necroinflammatory scores and the extent of fibrosis. GL and ω-3 significantly decreased liver malondialdehyde level (P < 0.005), liver NF-ƙB level (P < 0.005) and its tissue expression as detected by immunohistochemistry. In conclusion, glycyrrhizin and omega-3 fatty acids alone or in combination have potent anti-inflammatory, anti-oxidant and anti-fibrotic effects.

Ameliorative Effects of Soya Bean Oil and Vitamin C on Liver Enzymes in Ethanol -Induced Oxidative Stress in Wistar Rats

The protective potential of soya bean oil and vitamin C on Ethanol-induced oxidative stress in Wistar rats was evaluated. Thirty five Wistar rats of both sexes weighing 120-200g were grouped into five groups of seven rats each. Oxidative stress was induced by administration of 20% ethanol (w/v) for 28 days, at the end of the period of administration, the rats were fasted over night and their blood was collected directly from the heart via cardiac puncture in EDTA sample bottles and the blood was centrifuged and the serum was used to determine the levels of of AST, ALT and ALP in all the groups . The result revealed that Ethanol significantly (p<0.05) increased plasma AST, ALT and ALP levels. Vitamin C significantly (p<0.05) decreased serum AST while soya bean oil produced no significant (p<0.05) effect on plasma AST. The serum levels of ALT and ALP were significantly reduced following vitamin C and Soya bean oil administration. The elevation of serum AST, ALT and ALP after alcohol administration is an indication oxidative stress. The decrease in plasma concentration of the liver enzymes after treatment with vitamin C and Soya bean oil reveals the ameliorative potential of vitamin C and soya bean oil against ethanol-induced oxidative stress in Wistar rats.