摘要
目的研究白花丹醌对瘦素(leptin)刺激人肝星状细胞(HSC-LX2)增殖与α-平滑肌动蛋白(α-SMA)表达的影响。方法体外培养HSC-LX2,瘦素刺激24h,各药物组与细胞共孵育24h后,采用MTT法检测白花丹醌对HSC-LX2增殖的影响;采用荧光定量PCR检测各组α-SMAmRNA的表达和免疫组织化学方法检测α-SMA蛋白表达情况。结果白花丹醌呈剂量依赖性抑制瘦素诱导HSC-LX2增殖,以16μmol·L-1浓度时最为明显,与leptin对照组比较差异有显著性(P<0.01);白花丹醌各剂量组作用24h后,HSC-LX2细胞中α-SMAmRNA的水平降低,尤以中、高剂量组明显,与leptin组比较(P<0.01),且作用呈剂量依赖性。免疫组化结果显示白花丹醌各剂量组对HSC-LX2细胞α-SMA蛋白表达有一定的抑制作用。结论白花丹醌抗肝纤维化作用机制之一是从mRNA水平和蛋白水平抑制α-SMA表达,从而抑制HSCECM的合成,发挥抗肝纤维化作用。
Aim To investigate the effect of plumbagozeylanica(plumbago) on the expression of α-SMA in human hepatic stellate cells(HSC-LX2) activated by leptin.Methods HSC-LX2 were cultured in vitro,stimulated by leptin for 24 hours and treated with different concentrations of plumbago for 24 hours,before MTT assay was used to evaluate the inhibitive effect of plumbago.The expressions of α-SMA mRNA were determined by Real-time quantitative PCR.Results Different concentration of plumbago inhibited the activation and proliferation of HSC-LX2,in a dose-dependent manner.The inhibitive effect was most obvious in 16 μmol·L-1.The expressions of α-SMA mRNA were markedly reduced by the treatment with three concentrations of plumbago than that in the leptin group,especially in middle and high dose group(P 0.01),and was dose-dependent.Immunohistochemistry showed that different concentrations of plumbago also inhibited the expression of α-SMA to some extent.Conclusion The expressions of α-SMA can be down-regulated by plumbago,resulting in the reduced synthesis of ECM and the promoted degradation of ECM,which may be one of the possible molecular mechanism against hepatic fibrosis.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2010年第9期1154-1157,共4页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No30760321)
广西科学研究与技术开发项目(No桂科攻0992003-1)