白花丹醌对瘦素刺激的人肝星状细胞周期及其相关蛋白表达的影响  

Effects of plumbagin on cell cycle of human hepatic stellate cells stimulated by leptin and its related protein expression

作　　者：韦燕飞[1] 李景强[1] 张志伟[2] 张园[1] 王玖恒[1] 赵铁建[1] 

机构地区：[1]广西中医药大学基础医学院生理教研室,广西南宁530001 [2]广西中医药大学附属瑞康医院,广西南宁530001

出　　处：《中草药》2012年第9期1776-1780,共5页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(30760321);广西高等学校优秀人才资助项目(J11065);广西教育厅科研项目(200103YB083)

摘　　要：目的研究白花丹醌对瘦素刺激的体外培养人肝星状细胞(HSC-LX2)细胞周期及周期相关蛋白表达的影响,探讨白花丹醌抗肝纤维化的作用机制。方法 ELISA法检测HSC-LX2细胞培养上清液中自分泌瘦素水平。将HSC-LX2细胞分成对照组,瘦素组,白花丹醌2、8μmol/L组,秋水仙碱6.25μg/mL阳性对照组。除对照组外,其余各组细胞用瘦素100μg/mL刺激24 h、再与药物共同培养24 h后,流式细胞仪检测HSC-LX2细胞周期,Western blotting法检测细胞周期相关蛋白cyclin D1、cyclin E1、p21的表达。结果 HSC-LX2细胞自分泌瘦素的浓度先随着培养时间的延长而递减,24 h达到低谷值,48 h后又升至6 h水平,每个时间点分泌的瘦素量无显著差异(P>0.05)。与对照组相比,瘦素组HSC-LX2细胞增殖能力显著增强,S期和G2/M期细胞比例显著增加;白花丹醌2、8μmol/L干预HSC-LX2细胞后,G0/G1期细胞的比例明显提高,而S期和G2/M期细胞比例明显降低,且呈明显的浓度相关性。与对照组比较,瘦素组HSC-LX2细胞经瘦素刺激后细胞周期相关蛋白cyclin D1、cyclin E1的量显著增加,p21蛋白的表达受到抑制;白花丹醌2、8μmol/L和秋水仙碱明显降低cyclin Dl、cyclin E1蛋白水平,增强p21蛋白表达。结论白花丹醌可明显抑制瘦素诱导的HSC-LX2细胞增殖,该作用主要是通过抑制细胞由G0/G1期向S期转变而产生的,作用机制可能与其降低cyclin Dl、cyclin E1蛋白水平,增加p21蛋白表达相关。Objective To investigate the effects ofplumbagin on cell cycle of human hepatic satellite cells （HSC-LX2） stimulated by leptin in vitro and its expression of related proteins, so as to explore the mechanism of anti-hepatic fibrosis of plumbagin. Methods The level of autocrine leptin in culture supernate of HSC-LX2 was measured by ELISA. HSC-LX2 were divided into control, model （leptin）, plumbagin low- and high-dose （2 and 8 μmol/L）, and colchicines （6.25 μg/mL） groups. In addition to the control group, the other groups were stimulated by leptin （100 μg/mL） for 24 h and treated with plumbagin at different concentration and colchicine for 24 h later. HSC-LX2 cell cycle was detected by flow cytometry （FCM） and the expression of cell cycle-related proteins of cyclin D 1, cyclin E1, and p21 were tested by Western blotting. Results The concentration of autocrine leptin in HSC-LX2 decreased with cultural time prolonging, the minimum time was at 24 h and after 48 h the level recovered to original level of 6 h （P 〈 0.05）. There was no significant difference in concentration of autocrine leptin at each time point （P 〉 0.05）. FCM analysis showed that in model group HSC-LX2 cell proliferation and cell number ratio of S and G2/M phases significantly increased compared with the control group. At 24 h after addition ofplumbagin （2 and 8 μmol/L）, the percentage of HSC-LX2 at G0/G1 phase was apparent increased, while the total percentage of HSC-LX2 cells at S and G2/M phases was apparent decreased in a dose-dependent manner. Compared with the control group, the stimulation by leptin significantly increased the contents of cyclin DI and cyclin E1 in HSC-LX2, and inhibited the expression of p21 protein. However, plumbagin （2 and 8 μmol/L） and colchicine significantly decreased the level of cyclin D 1 and cyclin E1 and increased the expression of p21 protein. Conclusion Plumbagin could significantly inhibit the proliferation of HSC-LX2 induced by leptin through blocking HSC-LX2 to ent

关 键 词：白花丹醌 瘦素 人肝星状细胞 细胞增殖 细胞周期 细胞周期相关蛋白 

分 类 号：R285.5[医药卫生—中药学]