甘草甜素抗肝星状细胞增殖的相关机制研究(1)
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【摘要】目的:探讨甘草甜素抑制肝星状细胞增殖的相关机制。方法:用MTT法检测三组不同浓度甘草甜素对肝星状细胞增殖的抑制率,用免疫细胞化学法检测细胞周期素cyclinD1以及PCNA的表达情况。结果 MTT法检测不同药物浓度组:空白对照组、0.8mg/L 甘草甜素组、1.2mg/L甘草甜素组、1.6mg/L甘草甜素组细胞增殖抑制率分别为0,2.73%,14.75%,25.96%,有统计学意义(F=19.364,P<0.05);不同甘草甜素浓度组PCNA阳性表达细胞数与对照组比较差异有显著性(F=68.329,P<0.01);不同甘草甜素浓度组cyclinD1阳性表达细胞数与对照组比较差异有显著性(F=58.362,P<0.01)。结论:甘草甜素能够浓度依赖性的抑制大鼠肝星状细胞增殖;甘草甜素可通过降低细胞周期素cyclinD1的表达和PCNA的表达而抑制大鼠肝星状细胞增殖。
【关键词】甘草甜素;肝星状细胞;细胞周期素;PCNA ,cyclinD1
The Relevant Mechanisms Of Glycyrrhizin Inhibit Hepatic Stellate Cell Proliferation
WANG Xiaomei LIU JunmeiWANG JilingJI PengfeiZHU Lina
【Abstract】 ObjectiveTo discuss the relevant mechanisms of the glycyrrhizin inhibiting hepatic stellate cell proliferation. MethodeThe inhibition ratioes of the three group different density glycyrrhizin on hepatic stellatecell proliferation was detected by MTT method. The express of the cell cyclinD1 and PCNA was detected by immunocytochemical Method.RusultsThe inhibition ratio results of MTT method detecting different glycyrrhizin density groups inhibiting hepatic stellate cell proliferation: blank,0.8mg/l group,1.2mg/l group,1.6mg/l group in order were 0,2.73 percent,14.75 percent, 25.96 percent.It shows significant dose-dependent and have stati- stical significance (F=68.329,P<0.01).The cell population of PCNA positive express in the different glycyrrhizin groups have significant differences compared with control group (F=68.329, P<0.01). The cell population of cyclinD1 positive express in the different glycyrrhizin groups have significant differences compared with control group (F=58.362,P<0.01).ConclusionGlycyrrhizin relying on the density caninhibit the rat hepatic stellate cell proliferation. Glycyrrhizin may through cut down the cell cyclin express of cyclinD1 and PCNA inhibit the rat hepatic stellate cell proliferation.
【Key words】chronic liver disease,cytokine,IL-1,IL-6,TNF-α
【中图分类号】R248.2【文献标识码】B【文章编号】1005-0515(2010)010-0001-03
肝星状细胞(HSC)是细胞外基质(ECM)的主要来源细胞,在肝受到损伤时可被激活、转化及增殖,是肝纤维化发生、发展的核心环节。因此抑制HSC的活化和增殖是当今中医中药抗肝纤维化研究的一大热点。甘草甜素是由以β一甘草酸为主要成分的复方制剂,既往研究表明甘草甜素对肝星状细胞的增殖有抑制作用[1],但其对HSC活化、增值的抑制机制目前未见深入报道。本研究拟对肝星状细胞进行体外培养,从细胞周期调控角度探讨甘草甜素对肝星状细胞增殖的影响,进一步揭示肝纤维化的发生机制以及甘草甜素抑制肝星状细胞增殖的机制。
1 材料与方法
1.1 试验材料:肝星状细胞株HSC-T6,购自上海中医药大学肝病研究所。主要实验试剂有:特级胎牛血清(购自北京元亨圣马生物技术研究所),RPMI 1640 培养基、胰蛋白酶(购自Amresco),cyxlinD1抗体、PCNA抗体(购自Sant Cruz Biotechnoloyg),浓缩型DAB试剂盒:(购自北京中杉金桥生物技术有限公司),免疫组化染色试剂盒(购自北京中杉金桥生物技术有限公司),免疫染色一抗稀释液、免疫染色二抗稀释液以及封片液(购自江苏碧云天生物技术研究所) ......
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