Protective Effect of Water Extract of Citri Grandis Exocarpium on Alcohol-induced Acute Liver Injury / 中国实验方剂学杂志

ObjectiveTo explore the pharmacodynamic effect of the water extract of Citri Grandis exocarpium （WEC） on mice with alcohol-induced acute liver injury and provide data support for the development of this medicinal for anti-alcoholism and liver protection. MethodThe main components of WEC were determined by high performance liquid chromatography （HPLC）. Sixty Balb/c mice were randomized into 6 groups： control group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， model group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， low-， medium-， and high-dose WEC groups （0.5， 1.0， 2.0 g·kg-1）， and Haiwang Jinzun tablet positive control group （2.0 g·kg-1）. The administration lasted 14 days. One day before the end of the administration， mice were fasted for 12 h with free access to water. The mice， except the control group， were given 56° Chinese liquor （13 mL·kg-1）. After 2 h， blood was taken from eyeballs and the liver was dissected and weighed. Automatic biochemical analyzer was employed to detect the expression of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and alcohol dehydrogenase （ADH）. The pathological changes of liver tissues were observed based on hematoxylin-eosin （HE） staining， and apoptosis of hepatocytes based on TUNEL/DAB staining. The expression of proteins related to apoptosis was detected by Western blot. ResultAccording to the HPLC fingerprint， the main components of WEC were rhoifolin and naringin. Compared with the control group， the model group showed increase in liver/body weight ratio （P<0.01） and the expression of ALT and AST （P<0.05， P<0.01）， decrease in the expression of ADH （P<0.05）， blurred structure of hepatic lobules， pathological changes of liver tissue， loose cytoplasm with edema， severe steatosis， rise of the TUNEL-positive rate （P<0.01）， reduction in expression of Bcl-2 （P<0.01）， and increase in Bax and Caspase-3 （P<0.01）. Compared with the model group， medium-dose WEC lowered liver/body weight ratio （P<0.05）. All doses of WEC depressed the activity of ALT and AST （P<0.05， P<0.01）， up-regulated the expression of ADH （P<0.05）， significantly improved the pathological features of alcohol-induced cytoplasmic porosity， edema， and steatosis， down-regulated the TUNEL-positive rate （P<0.05， P<0.01）， enhanced the expression of Bcl-2 （P<0.05）， and decreased Bax and Caspase-3 （P<0.01）. ConclusionWEC regulates the expression of ALT， AST， and ADH and improves hepatic steatosis and hepatocyte apoptosis to fight against acute liver injury.

Protective Effect of Water Extract of Citri Grandis Exocarpium on Alcohol-induced Acute Liver Injury / 中国实验方剂学杂志

ObjectiveTo explore the pharmacodynamic effect of the water extract of Citri Grandis exocarpium （WEC） on mice with alcohol-induced acute liver injury and provide data support for the development of this medicinal for anti-alcoholism and liver protection. MethodThe main components of WEC were determined by high performance liquid chromatography （HPLC）. Sixty Balb/c mice were randomized into 6 groups： control group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， model group （equal volume of 0.5% carboxymethyl cellulose sodium solution）， low-， medium-， and high-dose WEC groups （0.5， 1.0， 2.0 g·kg-1）， and Haiwang Jinzun tablet positive control group （2.0 g·kg-1）. The administration lasted 14 days. One day before the end of the administration， mice were fasted for 12 h with free access to water. The mice， except the control group， were given 56° Chinese liquor （13 mL·kg-1）. After 2 h， blood was taken from eyeballs and the liver was dissected and weighed. Automatic biochemical analyzer was employed to detect the expression of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， and alcohol dehydrogenase （ADH）. The pathological changes of liver tissues were observed based on hematoxylin-eosin （HE） staining， and apoptosis of hepatocytes based on TUNEL/DAB staining. The expression of proteins related to apoptosis was detected by Western blot. ResultAccording to the HPLC fingerprint， the main components of WEC were rhoifolin and naringin. Compared with the control group， the model group showed increase in liver/body weight ratio （P<0.01） and the expression of ALT and AST （P<0.05， P<0.01）， decrease in the expression of ADH （P<0.05）， blurred structure of hepatic lobules， pathological changes of liver tissue， loose cytoplasm with edema， severe steatosis， rise of the TUNEL-positive rate （P<0.01）， reduction in expression of Bcl-2 （P<0.01）， and increase in Bax and Caspase-3 （P<0.01）. Compared with the model group， medium-dose WEC lowered liver/body weight ratio （P<0.05）. All doses of WEC depressed the activity of ALT and AST （P<0.05， P<0.01）， up-regulated the expression of ADH （P<0.05）， significantly improved the pathological features of alcohol-induced cytoplasmic porosity， edema， and steatosis， down-regulated the TUNEL-positive rate （P<0.05， P<0.01）， enhanced the expression of Bcl-2 （P<0.05）， and decreased Bax and Caspase-3 （P<0.01）. ConclusionWEC regulates the expression of ALT， AST， and ADH and improves hepatic steatosis and hepatocyte apoptosis to fight against acute liver injury.

FoxO1 regulates Hcyinduced hepatocyte apoptosis through ATF6 / 中国药理学通报

: Aim To explore the role and possible mechanism of forked transcription factor (FoxO1) in hepatocyte apoptosis induced by homocysteine (Hey) . Methods The male cbs

Uric Acid Induces Inflammation, Hepatocyte Apoptosis and Deterioration of Liver Function

@#Introduction: Uric acid is a common cause of liver tissue damage due to its hepatotoxic effect. This study is aimed to investigate: (1) the effect of uric acid on liver damage which can be seen from the serum levels of SGOT and SGPT, (2) the inflammatory response demonstrated by TLR-4 and MCP-1 mRNA expression, and (3) the proportion of hepatocytes apoptosis in mice. Methods: A total of 25 adult male Swiss-Webster mice were divided into five groups: one control group and four uric acid groups (AU7, AU14, AU21 and AU28). The uric acid groups were administered with 125 mg/kgBW uric acid for 7, 14, 21, and 28 days. Following the treatment, mice were terminated and the liver was harvested. Blood sample was taken from retro-orbital vein to assess serum uric acid, SGOT, and SGPT levels. RT-PCR was performed to examine the mRNA expressions of TLR-4 and MCP-1. TUNEL staining was used to assess the proportion of apoptotic hepatocytes. Results: Induction of uric acid caused hyperuricemia, increased expression of TLR-4 and MCP-1 mRNA significantly (p<0.05) which indicated an inflammatory reaction. The levels of SGOT and SGPT were elevated significantly (p<0.05), as well as the number of hepatocyte apoptosis (p<0.05). Conclusion: Hyperuricemia affected the inflammatory response by increasing the mRNA expression of TLR-4 and MCP-1. An increased number of apoptotic hepatocytes was likely caused by the ongoing inflammatory reaction during the induction of uric acid.

Swertiamarin ameliorates carbon tetrachloride-induced hepatic apoptosis via blocking the PI3K/Akt pathway in rats

Swertiamarin (STM) is an iridoid compound that is present in the Gentianaceae swertia genus. Here we investigated antiapoptotic effects of STM on carbon tetrachloride (CCl₄)-induced liver injury and its possible mechanisms. Adult male Sprague Dawley rats were randomly divided into a control group, an STM 200 mg/kg group, a CCl₄ group, a CCl₄+STM 100 mg/kg group, and a CCl₄+STM 200 mg/kg group. Rats in experimental groups were subcutaneously injected with 40% CCl₄ twice weekly for 8 weeks. STM (100 and 200 mg/kg per day) was orally given to experimental rats by gavage for 8 consecutive weeks. Hepatocyte apoptosis was determined by TUNEL assay and the expression levels of Bcl-2, Bax, and cleaved caspase-3 proteins were evaluated by western blot analysis. The expression of TGF-β1, collagen I, collagen III, CTGF and fibronectin mRNA were estimated by qRT-PCR. The results showed that STM significantly reduced the number of TUNEL-positive cells compared with the CCl₄ group. The levels of Bax and cleaved caspase-3 proteins, and TGF-β1, collagen I, collagen III, CTGF, and fibronectin mRNA were significantly reduced by STM compared with the CCl₄ group. In addition, STM markedly abrogated the repression of Bcl-2 by CCl₄. STM also attenuated the activation of the PI3K/Akt pathway in the liver. These results suggested that STM ameliorated CCl₄-induced hepatocyte apoptosis in rats.

Protective effect of earthworm active ingredients against endoplasmic reticulum stress induced L-02 hepatocyte apoptosis / 中国中药杂志

This study focused on the protective effect of earthworm active ingredients (EWAs) on hepatocyte apoptosis induced by endoplasmic reticulum stress (ERS) in L-02 cells. The L-02 cells were cultured in vitro. The cell viability was measured with CCK-8, the apoptosis of L-02 cells was detected by flow cytometry, and the relevant protein and mRNA expressions were detected by Western blot and qPCR. According to the findings, tunicamycin (TM) could obviously reduce the survival rate of L-02 cells in a time-dependent and dose-dependent manner. Compared with normal group, the apoptosis rate in model group was significantly increased (<0.05 or <0.01). The protein and mRNA expressions of ERS-related signal molecules, such as GRP78, PERK, eLF2α, ATF4, CHOP and Bax, were significantly up-regulated (<0.05 or <0.01), while Bcl-2 was significantly down-regulated (<0.05 or <0.01). After the administration with different concentrations of EWAs, compared with model group, EWAs could significantly increase the survival rate ofL-02 hepatocyte and decrease the cell apoptosis rates. It could also reduce the protein and mRNA expressions of ERS-related signal molecules, such as GRP78, PERK, eLF2α, ATF4, CHOP and Bax, in a dose-dependent manner (<0.05 or <0.01) and increased the protein and mRNA expressions of Bcl-2(<0.05 or <0.01). These results showed that EWAs had a significantly protective effect on hepatocyte apoptosis induced by ERS in L-02 cells. Its mechanism may be related to the down-regulation of mRNA and protein expressions of GRP78, PERK, ATF4, eLF2α, CHOP and Bax, and the up-regulation, the relief of ERS and the promotion of the proliferation of impaired L-02 cells.

Liraglutide ameliorates liver injury of type 2 diabetic mice via micro-RNA-33-AMPK pathway / 中国病理生理杂志

AIM:To observe the effects of liraglutide on the level of microRNA-33 (miR-33) and the expres-sion of AMP-activated protein kinase ( AMPK ) and apoptosis-related proteins in mice with type 2 diabetes mellitus (T2DM), and to explore its possible mechanism .METHODS:High-fat diet and intraperitoneal injection of streptozocin were used to establish the type 2 diabetic model in C57BL/6 mice.The mice were randomly divided into 4 groups ( n=15 ):in control group , the normal mice were subcutaneously injected with equivalent volume of saline ;in model group , the T2DM mice were subcutaneously injected with equivalent volume of saline ; in low-and high-dose liraglutide treatment groups, the T2DM mice were subcutaneously injected with 100 and 200 μg? kg -1? d-1, respectively.After 4 weeks of administration, the levels of FBG, TG, TC, HDL-C, LDL-C, ALT and AST were determined.HE staining was used to ob-serve the pathological changes of the liver tissues .The protein level of cleaved caspase-3 in the liver tissue was detected by the technique of immunofluorescence .The protein levels of p-AMPK/AMPK and apoptosis-related proteins were detected by Western blot .The expression of miR-33 in the liver tissues was detected by real-time PCR.RESULTS: Compared with model group, the contents of FBG, TG, TC, LDL-C, ALT and AST were decreased significantly , while the content of HDL-C was increased significantly in low-dose liraglutide group and high-dose liraglutide group ( P<0.05 ) .The protein levels of phosphorylated AMPK and Bcl-2 were up-regulated significantly , and the expression of cleaved caspase-3 was down-regulated significantly (P<0.05).The level of miR-33 was decreased significantly (P<0.01).CONCLUSION:Liraglutide alleviates liver injury in type 2 diabetic mice , and the mechanism may be associated with reducing the level of miR-33 and increasing the phosphorylation of AMPK in the liver tissues , thereby inhibiting hepatocyte apoptosis .

Effect of nodosin on accelerating hepatocyte regeneration after partial liver transplantation in rats / 器官移植

Objective To evaluate the effect of nodosin,as an effective element extracted from rabdosiae serrae, on hepatocyte regeneration after partial liver transplantation.Methods Wistar rats were used as donors and SD rats as recipients.Rat models with partial liver transplantation were established by modified two-cuff technique.Twenty-four recipient rats were randomly assigned into the nodosin and control groups.In the nodosin group,nodosin at a dosage of 1 00 μg/ml was administered via tail venous route after liver transplantation.Peripheral plasma and liver specimen were obtained at postoperative 3 and 7 d.The levels of alanine transaminase (ALT),aspartate aminotransferase (ALT)and albumin (ALB)in the peripheral plasma were measured by spectrophotometry.Hepatic histomorphological changes were observed under light microscope.The positive cell count of proliferating cell nuclear antigen (PCNA)antibody in the liver tissue was detected by immunohistochemistry. The expression levels of phosphorylated protein kinase (p-AKT ), phosphorylated mammalian target of rapamycin (p-mTOR),cyclin D1 and heme oxygenase (HO)-1 proteins were measured by western blot.The apoptosis of liver cells was detected by Annexin V method and TdT mediated-dUTP nick end labeling (TUNEL).Results Compared with the control group,the serum levels of ALT and AST were significantly lower at 3 d and 7 d after operation,whereas the ALB content was significantly higher in the nodosin group (all in P <0.05).And nodosin could alleviate the pathological injury of rat liver tissue after transplantation.The positive cell count of PCNA in the nodosin group was significantly higher than that in the control group (P <0.05).In the nodosin group,the expression levels of p-AKT,p-mTOR,cyclin D1 and HO-1 proteins were significantly higher than those in the control group (all in P <0.05).The quantity and percentage of apoptotic hepatocytes in the nodosin group were significantly lower than those in the control group (both in P <0.05).Conclusions Application of nodosin can decrease the quantity of apoptotic hepatocytes and accelerate hepatocyte proliferation after liver transplantation in rat models.

Study on mechanism of NOXs in liver fibrosis / 中国药理学通报

Nicotinamide adenine dinucleotide phosphate oxidase ( NOXs) contributes to the production of reactive oxygen species ( ROS) in liver fibrosis, resulting in the activation of endoplas-mic reticulum stress ( ERS ) and IRE1α-XBP1 signaling path-way. ROS is a series of oxygen metabolites and its derivatives, produced by the single electron reduction of molecular oxygen ( O2 ) , including superoxide anion ( O2- ) , hydroxyl radical (-OH) , hydrogen peroxide ( H2 O2 ) , hypochlorite ion ( OCl-) and so on. They can interact with a large number of molecules, including small inorganic molecules, proteins, lipids, carbohy-drates and nucleic acids, resulting in lipid peroxidation of cell damaging molecules. And as a second messenger, ROS can also affect the proliferation and activation of HSC in liver fibrosis, and induce the hepatocyte apoptosis through a variety of cellular signal transduction. Here we review the current status of the study on the mechanism of NOXs in liver fibrosis.

Endoplasmic reticulum stress and non-alcoholic fatty liver disease / 国际药学研究杂志

The“two-hit”theory which underlies the mechanism of non-alcoholic fatty liver disease (NAFLD) is difficult to explain the progress of NAFLD from liver lipid accumulation to steatohepatitis, liver fibrosis and the gradually aggravated diseases.Recently the“multiple-hit”theory emphasizes the significance of endoplasmic reticulum stress (ERS) in the progression of NAFLD. The endoplasmic reticulum is a cytoplasmic organella for protein and lipid synthesis. ERS, which can be induced by the unbalance of endoplasmic reticulum homeostasis, plays a critical role in hepatocyte lipid metabolic disorders, Apoptosis and the progression of NAFLD. In this paper, the relevance of ERS and non-alcoholic liver lipid accumulation, steatohepatitis and hepatocyte apoptosis is reviewed.

Role of TNF-? in the mechanism of hepatocyte apoptosis induced by intestinal perforations due to abdominal firearm wound / 中国现代普通外科进展

Objective:To study the role of TNF-?in the mechanism of hepatocyte apoptosis induced by intestinal perforations due to abdominal firearm wound. Methods:A total of 42 Chang-Bai piglets were divided randomly into 7 groups: control group and wounded 1, 2 , 4, 8, 12, 24 hour groups.The model of intestinal perforations due to abdominal firearm wound were established in wounded groups. Levels of plasma endotoxin were measured using ehromogenic limulus amehoeyte lysate test.Hepatic TNF-? content was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum TNF-? levels were determined at the same time. Results: Levels of plasma endotoxin, serum TNF-?, hepatic TNF-? content and hepatocyte apoptosis indexes in wounded groups were all significantly elevated compared with control group(P

Reduced apoptosis induced by endotoxin in mice blocking TRAIL with soluble death receptor 5 / 中华微生物学和免疫学杂志

Objective To explore the effect of TNF related apoptosis inducing ligand (TRAIL) in apoptosis induced by LPS. Methods After LPS injected mice blocking TRAIL with soluble death receptor 5 (sDRS), detecting ALT, AST and LDH of mice serum at different times, apoptotic effects of LPS to mice hepatocyte were detected by HE and flow eytometry (FCM) with Annexin V-FITC/PI staining. The expres-sion of DR5 in mice hepatocyte was assayed with immunohistochemistry and FCM. Results Apoptotic effect was promoted by up-regulated DR5 expression on hepatocyte. Blocking TRAIL with sDR5 markedly amelio-rated the hepatocyte damage and reduced apoptosis. Conclusion These results establish a critical role for TRAIL in apoptosis during disease process of LPS.

Role of NF-?B in hepatocyte apoptosis induced by intestinal perforations due to abdominal firearm wound / 中国现代普通外科进展

Objective: To investigate the role of NF-?B in signal transduction of hepatocyte apoptosis in liver injury. Methods: A total of 42 Chang-Bai piglets were divided into 7 groups: control group, 1, 2, 4, 8, 12, and 24 hours wound group. The model of intestinal perforations due to abdominal firearm wound was established in wound groups. Hepatic NF-?B activity was measured with immunohistochemical staining and image analysis in all groups. Hepatocyte apoptosis indexes and serum ALT levels were also determined. Results: Levels of hepatic NF-?B activity in wounded groups were significantly elevated compared with control group, and there were two peaks (1 and 8 hours group P

The Relationship Between the Apoptosis Hepatocyte and Its Genic Mediation and the Ischemia of Portal Vein / 中国普外基础与临床杂志

Objective To introduce the relationship between the apoptosis hepatocyte and its genic mediation and the ischemia of portal vein. Methods The combination of related literatures and our research findings were made.Results Portal vein ischemia may induced hepatocyte apoptosis, p53 and bcl-2 gene alternatively adjust hepatocyte apoptosis. Expression of p53 gene is enhanced in hepatic tissue when hepatocyte apoptosis is not obvious, but after 24-72 h of portal vein ischemia, when hepatocyte apoptosis is obvious, enhanced expression of p53 gene or reduced expression of bcl-2 gene occur. There exists close relationship between portal vein ischemia and hepatocyte apoptosis. Conclusion Apoptosis hepatocyte is involved in organic atrophy after ischemia of portal vein, and p53 and bcl-2 gene alternatively adjust hepatocyte apoptosis. At present, the mechanism of apoptosis of hepatocyte induced by ischemia of portal vein is not clear, which needs further study.

Intrathymic inoculation of liver specific antigen protects hepatocytes from apoptosis after liver allotransplantation / 中国病理生理杂志

AIM: To study the effects of intrathymic inoculation of liver specific antigen (LSA) on hepatocyte apoptosis after liver allotransplantation. METHODS: Orthotopic liver transplantation was used in this study. Group Ⅰ: syngenic control (Wistar-to-Wistar); Group Ⅱ: acute rejection (SD-to-Wistar); Group Ⅲ: thymus inoculation of SD rat LSA day 7 before transplantation. The observation of general situation and survival time, hepatocyte apoptosis and LAT expression in liver transplants were used to analyze immune state of animals in different groups. RESULTS: The general situation of group Ⅰ was very well after transplantation. Recipients of groupⅡ lost body weight progressively and all died within day 9 to day 13 post transplantation. As for group Ⅲ, the general situation of recipients was remarkably better than that in group Ⅱ. The positive cells of apoptosis in group Ⅲ detected by TUNEL were not significantly different from that in group Ⅰ, but was significantly lower than that in group Ⅱ. LAT was detected at any time in group Ⅱ with peak expression at day 5 and day 7 post transplantation. In contrast, LAT was not detected in any other groups. CONCLUSION: Intrathymic inoculation of LSA protects hepatocytes from apoptosis after liver allotransplantation.