Effects of asiatic acid on the expression of mRNA of smad7 and PPARγ in HSC-T6 in vitro
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摘要: 目的检测积雪草酸对HSC-T6细胞Smad-7和PPARγ的mRNA表达的影响,探讨积雪草酸抗肝纤维化的分子生物学机制。方法用MTT实验检测积雪草酸对HSC-T6细胞的增殖抑制作用,计算抑制率,确定积雪草酸作用浓度;设置空白对照组与积雪草酸干预组,用半定量RT-RCR检测两组HSC-T6细胞Smad7和PPARγ的mRNA表达差异。结果积雪草酸抑制HSC-T6细胞的增殖,药物浓度为10、20、30、40、50、70、90μmol/L时,抑制率分别为7.47%、12.42%、13.97%、21.32%、47.31%、51.18%、60.36%。空白对照组和积雪草酸干预组的半定量RT-PCR的结果分别为:Smad7/GAPDH mRNA空白对照组为0.267±0.037,积雪草酸干预组为0.358±0.035;PPARγ/GAPDH mRNA空白对照组为0.157±0.054,积雪草酸干预组为0.570±0.066。两组结果间比较差异具有统计学意义,P<0.05。结论积雪草酸可以上调HSC-T6细胞Smad7和PPARγ的mRNA的表达,从而抑制HSC-T6细胞合成和分泌胶原,改善肝...Abstract: Objective To investigate the effects of Asiatic acid on the expression of mRNA of smad7 and PPARγ in HSC-T6 in vitro and to explore the molecular mechanism of the anti-liver fibrosis activity of the Asiatic acid.Methods The influence of asiatic acid on HSC proliferation was detected by MTT assay and optimum concentration was determined.The cultured HSC were divided into control group and Asiatic acid treatment group.The influence of asiatic acid on the expression of mRNA of smad7 and PPARγ in HSC-T6 in vitro was detected by semi-quantitative RT-PCR.Results The asiatic acid significantly inhibits the proliferation of HSC in a dose-dependent manner.Compared with the control group, the expression of mRNA of smad7 and PPARγ of HSC-T6 in vitro in the Asiatic acid treatment group significantly increased (samd7/GAPDHmRNA:0.267±0.037vs0.358±0.035;PPARγ/GAPDHmRNA:0.157±0.054 vs 0.570±0.066) .There was significant difference between the two groups P<0.05.Conclusion Asiatic acid inhibits the collagen synthesis and secretion in HSC and improves the pathology of liver fibrosis possibly by up-regulating the expression of mRNA of smad7 and PPARγ.
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Key words:
- hepatocytes /
- Smad7 protein /
- PPAR gamma /
- RNA /
- messenger
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[1]杨光, 谭家驹, 何瑞华, 等.积雪草酸阻断大鼠肝纤维化的实验研究[J].中国中医基础医学杂志, 2009, 15 (07) :540-542. [2]Vogel S, Piantedosi R, Frank J, et al.An immortalized rat liver stellate cell line (HSC-T6) :a new cell model for the study of retinoid metabolism in vitro[J].J Lipid Res, 2000, 41 (6) :882-893. [3]汤丽霞, 杨光, 谭家驹.积雪草酸抑制HSC-T6细胞Ⅰ型胶原蛋白表达[J].第四军医大学学报, 2007, 28 (13) :1178-1180. [4]汤丽霞, 杨光, 谭家驹.积雪草酸诱导大鼠肝星状细胞凋亡[J].中草药, 2009 (40) :230-232. [5]Friedman SL.Molecular regulation of hepatic fibrosis, an integrated cellular response to tissue injury[J].J Biol Chem, 2000, 275 (4) :2247-2250. [6]Elsharkawy AM, Oakley F, Mann DA.The role and regulation of hepatic stellate cell apoptosis in reversal of liver fibrosis[J].Apoptosis, 2005, 10 (5) :927-939. [7]Friedman SL.Cytokines and fibrogenesis[J].Semin Liver Dis, 1999, 19 (2) :129-140. [8]Dooley S, Delvoux B, Lahme B, et al.Modulation of transforming growth factor beta response and signaling during transdifferentiation of rat hepatic stellate cells to myofibroblasts[J].Hepatology, 2000, 31 (5) :1094-1106. [9]Yan X, Liu Z, Chen Y.Regulation of TGF-beta signaling by Smad7[J].Acta Biochim Biophys Sin (Shanghai) , 2009, 41 (4) :263-272. [10]Tang LX, Yang G, Tang JJ.Smad7 inhibits collagen expression in human hepatic satellite cells in vitro[J].Nan Fang Yi Ke Da Xue Xue Bao, 2009, 29 (10) :2122-2123, 2127. [11]Dooley S, Hamzavi J, Ciuclan L, et al.Hepatocyte-specific Smad7 expression attenuates TGF-beta-mediated fibrogenesis and protects against liver damage[J].Gastroenterology, 2008, 135 (2) :642-659. [12]潘姝, 利天增, 李叶扬, 等.积雪草甙对增生性瘢痕成纤维细胞增殖与Smad7蛋白的影响[J].第三军医大学学报, 2005, 27 (1) :49-52. [13]Miyahara T, Schrum L, Rippe R, et al.Peroxisome pro-liferator-activated receptors and hepatic stellate cell activation[J].J Biol Chem, 2000, 275 (46) :35715-35722. [14]Marra F, Efsen E, Romanelli RG, et al.Ligands of peroxisome proliferator-activated receptor gamma modulate profibrogenic and proinflammatory actions in hepatic stellate cells[J].Gastroenterology, 2000, 119 (2) :466-478. [15]Gressner OA, Gressner AM.Connective tissue growth factor:a fibrogenic master switch in fibrotic liver diseases[J].Liver Int, 2008, 28 (8) :1065-1079. [16]Sun K, Wang Q, Huang XH.PPAR gamma inhibits growth of rat hepatic stellate cells and TGF beta-induced connective tissue growth factor expression[J].Acta Pharmacol Sin, 2006, 27 (6) :715-723. [17]Galli A, Crabb D, Price D, et al.Peroxisome proliferator-activated receptor gamma transcriptional regulation is involved in platelet-derived growth factor-induced proliferation of human hepatic stellate cells[J].Hepatology, 2000, 31 (1) :101-108. [18]Oakley F, Meso M, Iredale JP, et al.Inhibition of inhibitor of kappaB kinases stimulates hepatic stellate cell apoptosis and accelerated recovery from rat liver fibrosis[J].Gastroenterology, 2005, 128 (1) :108-120. [19]Straus DS, Pascual G, Li M, et al.15-deoxy-delta 12, 14-prostaglandin J2 inhibits multiple steps in the NF-kappa B signaling pathway[J].Proc Natl Acad Sci U S A, 2000, 97 (9) :4844-4849. [20]Zhao C, Chen W, Yang L, et al.PPAR gamma agonists prevent TGF beta1/Smad3-signaling in human hepatic stellate cells[J].Biochem Biophys Res Commun, 2006, 350 (2) :385-391.
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