人参皂苷Rg1调节NF-κB/VCAM-1减轻胆汁淤积性肝纤维化
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摘要
本文通过胆总管结扎(bile duct ligation, BDL)建立胆汁淤积性肝纤维化模型小鼠,研究人参皂苷Rg1对胆汁淤积诱发的肝纤维化的治疗作用,并探讨潜在的作用机制。本文中所有动物实验都获得单位伦理学委员会批准。检测血清生化指标和病理切片评估小鼠肝功能、肝损伤和纤维化;免疫组织化学和qPCR等方法检测血管细胞黏附分子-1 (vascular cell adhesion molecule 1, VCAM-1)在BDL诱导的肝纤维化过程中的表达变化规律;为探讨Rg1治疗作用的相关机制,检测转录因子-κB (nuclear factor-κB, NF-κB)及炎症因子。结果显示, VCAM-1表达先上调,至第7天达到高峰,随后表达有所下降,但与假手术组相比仍有高表达。与模型组相比, 40 mg·kg~(-1)·d~(-1)Rg1治疗可降低血清中谷草转氨酶(AST)、谷丙转氨酶(ALT)和总胆红素(T.Bili)水平(P<0.05或P<0.01),减轻肝功能损害,减缓BDL诱导的肝纤维化进程,显著地下调VCAM-1蛋白表达(P<0.05),抑制炎症反应。此外, Rg1可明显降低细胞核内NF-κB p65蛋白水平(P<0.05)。本研究表明VCAM-1在BDL诱导的肝纤维化过程中的表达呈动态变化, Rg1通过调节NF-κB/VCAM-1信号途径抑制炎症损伤,减轻胆汁淤积性肝纤维化,为将来Rg1开发成有效的肝纤维化治疗药物提供实验基础。
A mouse model of cholestatic liver fibrosis was established by bile duct ligation(BDL) method.The effect of ginsenoside Rg1 in the disease progress and the mechanism of cholestatic liver fibrosis are investigated in this mouse model. All animal experiments in this paper have been approved by the Unit Ethics Committee.Analysis of serum biochemical indicators and pathological sections assessed liver function, liver damage and fibrosis in mice. Immunohistochemistry and Western blot assays were used to detect vascular cell adhesion molecule-1(VCAM-1) in BDL-induced mice. Nuclear factor-κB(NF-κB) and inflammatory factors were detected to investigate related mechanism of Rg1. The results showed that expression of VCAM-1 was up-regulated and peaked at 7 days, followed by decreased expression, but still efficiently expressed compared to the sham-operated group. Compared with the model group, 40 mg · kg~(-1)· d~(-1)Rg1 treatment reduced serum aspartate transaminase(AST), alanine transaminase(ALT) and total bilirubin(T.Bili) levels(P<0.05 or P<0.01) and liver function damage,alleviated BDL-induced liver fibrosis, significantly down-regulated the expression of VCAM-1(P<0.05), and inhibited the inflammatory response. In addition, Rg1 significantly reduced NF-κB p65 level in the cellular nucleus(P<0.05). This study demonstrates that VCAM-1 is dynamically altered during BDL-induced liver fibrosis. Rg1 could dampen inflammation and alleviate cholestatic liver fibrosis via regulation of the NF-κB/VCAM-1 pathway.The results provide an experimental basis for Rg1 application for treating liver fibrosis.
A mouse model of cholestatic liver fibrosis was established by bile duct ligation(BDL) method.The effect of ginsenoside Rg1 in the disease progress and the mechanism of cholestatic liver fibrosis are investigated in this mouse model. All animal experiments in this paper have been approved by the Unit Ethics Committee.Analysis of serum biochemical indicators and pathological sections assessed liver function, liver damage and fibrosis in mice. Immunohistochemistry and Western blot assays were used to detect vascular cell adhesion molecule-1(VCAM-1) in BDL-induced mice. Nuclear factor-κB(NF-κB) and inflammatory factors were detected to investigate related mechanism of Rg1. The results showed that expression of VCAM-1 was up-regulated and peaked at 7 days, followed by decreased expression, but still efficiently expressed compared to the sham-operated group. Compared with the model group, 40 mg · kg~(-1)· d~(-1)Rg1 treatment reduced serum aspartate transaminase(AST), alanine transaminase(ALT) and total bilirubin(T.Bili) levels(P<0.05 or P<0.01) and liver function damage,alleviated BDL-induced liver fibrosis, significantly down-regulated the expression of VCAM-1(P<0.05), and inhibited the inflammatory response. In addition, Rg1 significantly reduced NF-κB p65 level in the cellular nucleus(P<0.05). This study demonstrates that VCAM-1 is dynamically altered during BDL-induced liver fibrosis. Rg1 could dampen inflammation and alleviate cholestatic liver fibrosis via regulation of the NF-κB/VCAM-1 pathway.The results provide an experimental basis for Rg1 application for treating liver fibrosis.
引文
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[15]Yao X,Jiang W,Ma CH,et al.Protective effects of ginsenoside Rg1 against carbon tetrachloride-induced liver injury in mice through suppression of inflammation[J].Phytomedicine,2016,23:583-588.
[16]Dong XQ,Duan LP,Miao YL,et al.A stereological analysis of ginsenoside Rg1 in inhibiting hepatic fibrosis[J].Chongqing Med(重庆医学),2014,43:3599-3601,3616.
[17]Geng J,Peng W,Huang Y,et al.Ginsenoside-Rg1 from Panaxnotoginseng prevents hepatic fibrosis induced by thioacet‐amidein rats[J].Eur J Pharmacol,2010,634:162-169.
[18]Kirkland JG,Godfrey CB,Garrett R,et al.Reversible surgical model of biliary inflammation and obstructive jaundice in mice[J].J Surg Res,2010,164:221-227.
[19]Friedman SL.Mechanisms of hepatic fibrogenesis[J].Gastroen‐terology,2008,134:1655-1669.
[20]Volpes R,van den Oord JJ,Desmet VJ.Immunohistochemical study of adhesion molecules in liver inflammation[J].Hepatology,1990,12:59-65.
[21]Xie CL,Wang WW,Xue XD,et al.A systematic review and meta-analysis of ginsenoside-Rg1(G-Rg1)in experimental ischemic stroke[J].Sci Rep,2015,5:7790.
[22]Ghonem NS,Assis DN,Boyer JL.Fibrates and cholestasis[J].Hepatology,2015,62:635-643.
[23]Lee UE,Friedman SL.Mechanisms of hepatic fibrogenesis[J].Best Pract Res Clin Gastroenterol,2011,25:195-206.
[24]Zhang D,Yuan D,Shen J,et al.Up-regulation of VCAM1relates to neuronal apoptosis after intracerebral hemorrhage in adult rats[J].Neurochem Res,2015,40:1042-1052.
[25]Yamada Y,Arao T,Matsumoto K,et al.Plasma concentrations of VCAM-1 and PAI-1:a predictive biomarker for post-operative recurrence in colorectal cancer[J].Cancer Sci,2010,101:1886-1890.
[26]Lo Iacono O,Rincón D,Hernando A,et al.Serum levels of soluble vascular cell adhesion molecule are related to hyperdynamic circulation in patients with liver cirrhosis[J].Liver Int,2008,28:1129-1135.
[27]Afford SC,Humphreys EH,Reid DT,et al.Vascular cell adhesion molecule 1 expression bybiliaryepithelium promotes persistence of inflammation by inhibiting effector T-cell apoptosis[J].Hepatology,2014,59:1932-1943.
[28]Agassandian M,Tedrow JR,Sembrat J,et al.VCAM-1 is a TGF-β1 inducible gene upregulated in idiopathic pulmonary fibrosis[J].Cell Signal,2015,27:2467-2473.
[29]Chu SF,Zhou X,Zhang Z,et al.HMGB1 is involved into the anti-ischemic stroke effect of ginsenoside Rg1 in diabetic rats[J].Acta Pharm Sin(药学学报),2018,53:958-965.
[30]Marx N,Sukhova GK,Collins T,et al.PPARαactivators inhibit cytokine-induced vascular cell adhesion molecule-1 expression in human endothelial cells[J].Circulation,1999,99:3125-3131.
[2]Davis GL,Alter MJ,El-Serag H,et al.Aging of hepatitis C virus(HCV)-infected persons in the United States:a multiple cohort model of HCV prevalence and disease progression[J].Gastroen‐terology,2010,138:513-521.e6.
[3]Mitchell AE,Colvin HM,Beasley RP.Institute of medicine recommendations for the prevention and control of hepatitis Band C[J].Hepatology,2010,51:729-733.
[4]Fujii H,Kawada N.Fibrogenesis in alcoholic liver disease[J].World J Gastroenterol,2014,20:8048-8054.
[5]Hernandez-Gea V,Friedman SL.Pathogenesis of liver fibrosis[J].Annu Rev Pathol,2011,6:425-456.
[6]Budiarto BR,Chan WH.Oxidative stresses-mediatedapoptotic effects of ginsenoside Rb1 on pre-and post-implantation mouse embryos in vitro and in vivo[J].Environ Toxicol,2017,32:1990-2003.
[7]Tian W,Chen L,Zhang L,et al.Effects of ginsenoside Rg1 on glucose metabolism and liver injury in streptozotocin-induced type 2 diabetic rats[J].Genet Mol Res.2017.DOI:10.4238/gmr16019463.
[8]Luo P,Chu SF,Gao Y,et al.Advances in study of pharmacologi‐cal effects of ginsenoside Rg1 in hepatic diseases[J].Acta Pharm Sin(药学学报),2018,53:21-27.
[9]Gao Y,Chu SF,Li JW,et al.Anti-inflammatory function of ginsenoside Rg1 on alcoholic hepatitis through glucocorticoid receptor related nuclear factor-kappa B pathway[J].J Ethnophar‐macol,2015,173:231-240.
[10]Gao Y,Chu SF,Xia CY,et al.Rg1 attenuates alcoholic hepatic damage through regulating AMPK and Nrf2 signal pathways[J].J Asian Nat Prod Res,2016,18:765-778.
[11]Gao Y,Chu SF,Shao QH,et al.Antioxidant activities of ginsen‐oside Rg1 against cisplatin-induced hepatic injury through Nrf2signaling pathway in mice[J].Free Radic Res,2017,51:1-13.
[12]Gao Y,Chu S,Zhang Z,et al.Hepataprotective effects of ginsen‐oside Rg1-a review[J].J Ethnopharmacol,2017,206:178-183.
[13]Zhao J,Shi Z,Liu S,et al.Ginsenosides Rg1 from Panaxgin‐seng:a potential therapy for acute liver failure patients?[J].Evid Based Complement Alternat Med,2014,2014:538059.
[14]Li JP,Gao Y,Chu SF,et al.Nrf2 pathway activationcontributes to anti-fibrosis effects of ginsenoside Rg1 in arat model of alcoholand CCl4-induced hepatic fibrosis[J].Acta Pharmacol Sin,2014,35:1031-1044.
[15]Yao X,Jiang W,Ma CH,et al.Protective effects of ginsenoside Rg1 against carbon tetrachloride-induced liver injury in mice through suppression of inflammation[J].Phytomedicine,2016,23:583-588.
[16]Dong XQ,Duan LP,Miao YL,et al.A stereological analysis of ginsenoside Rg1 in inhibiting hepatic fibrosis[J].Chongqing Med(重庆医学),2014,43:3599-3601,3616.
[17]Geng J,Peng W,Huang Y,et al.Ginsenoside-Rg1 from Panaxnotoginseng prevents hepatic fibrosis induced by thioacet‐amidein rats[J].Eur J Pharmacol,2010,634:162-169.
[18]Kirkland JG,Godfrey CB,Garrett R,et al.Reversible surgical model of biliary inflammation and obstructive jaundice in mice[J].J Surg Res,2010,164:221-227.
[19]Friedman SL.Mechanisms of hepatic fibrogenesis[J].Gastroen‐terology,2008,134:1655-1669.
[20]Volpes R,van den Oord JJ,Desmet VJ.Immunohistochemical study of adhesion molecules in liver inflammation[J].Hepatology,1990,12:59-65.
[21]Xie CL,Wang WW,Xue XD,et al.A systematic review and meta-analysis of ginsenoside-Rg1(G-Rg1)in experimental ischemic stroke[J].Sci Rep,2015,5:7790.
[22]Ghonem NS,Assis DN,Boyer JL.Fibrates and cholestasis[J].Hepatology,2015,62:635-643.
[23]Lee UE,Friedman SL.Mechanisms of hepatic fibrogenesis[J].Best Pract Res Clin Gastroenterol,2011,25:195-206.
[24]Zhang D,Yuan D,Shen J,et al.Up-regulation of VCAM1relates to neuronal apoptosis after intracerebral hemorrhage in adult rats[J].Neurochem Res,2015,40:1042-1052.
[25]Yamada Y,Arao T,Matsumoto K,et al.Plasma concentrations of VCAM-1 and PAI-1:a predictive biomarker for post-operative recurrence in colorectal cancer[J].Cancer Sci,2010,101:1886-1890.
[26]Lo Iacono O,Rincón D,Hernando A,et al.Serum levels of soluble vascular cell adhesion molecule are related to hyperdynamic circulation in patients with liver cirrhosis[J].Liver Int,2008,28:1129-1135.
[27]Afford SC,Humphreys EH,Reid DT,et al.Vascular cell adhesion molecule 1 expression bybiliaryepithelium promotes persistence of inflammation by inhibiting effector T-cell apoptosis[J].Hepatology,2014,59:1932-1943.
[28]Agassandian M,Tedrow JR,Sembrat J,et al.VCAM-1 is a TGF-β1 inducible gene upregulated in idiopathic pulmonary fibrosis[J].Cell Signal,2015,27:2467-2473.
[29]Chu SF,Zhou X,Zhang Z,et al.HMGB1 is involved into the anti-ischemic stroke effect of ginsenoside Rg1 in diabetic rats[J].Acta Pharm Sin(药学学报),2018,53:958-965.
[30]Marx N,Sukhova GK,Collins T,et al.PPARαactivators inhibit cytokine-induced vascular cell adhesion molecule-1 expression in human endothelial cells[J].Circulation,1999,99:3125-3131.