丹参单体通过下调粘着斑激酶诱导H_2O_2刺激的肝星状细胞凋亡  

Induction of apoptosis with Salvia miltiorrhiza monomer IH764-3 via downregulating focal adhesion kinase in H_2O_2-stimulated hepatic stellate cells

作　　者：姜慧卿[1] 张晓岚[1] 刘丽[1] 

机构地区：[1]河北医科大学第二医院消化科,河北石家庄050000

出　　处：《中国病理生理杂志》2003年第1期18-21,共4页Chinese Journal of Pathophysiology

基　　金：河北省科技攻关项目 (No .0 1 2 761 34);河北省自然科学基金资助项目 (No.30 1 361 )

摘　　要：目的 :研究丹参单体IH764 - 3对H2 O2 刺激的肝星状细胞 (HSCs)凋亡的诱导作用以及此过程中粘着斑激酶 (FAK)的变化。方法 :通过直接细胞计数法测定HSCs增殖 ;光学显微镜及透射电镜技术观察凋亡细胞的形态学改变 ,并应用膜联蛋白 (Annexin -V) /碘化丙啶 (PI)联合标记法检测HSCs的凋亡率 ;运用RT -PCR方法观察FAKmRNA表达。结果 :H2 O2 具有刺激HSCs增殖的作用 ;丹参单体IH764 - 3能够诱导H2 O2 刺激的HSCs凋亡 ,并呈剂量依赖关系 :1 0mg/L、2 0mg/L、30mg/L及 40mg/LIH764 - 3干预 48h后各组凋亡率分别为 6 35 %、9 2 8%、1 5 1 0 %、1 9 69% ,而H2 O2 组为 2 30 % ;30mg/LIH764 - 3干预HSCs不同时间 (1 2h、2 4h、48h)的凋亡率分别是6 73 %、1 0 34 %、1 5 1 0 % ,呈时间依赖关系 ;RT -PCR分析表明FAK基因表达强度在加入IH764 - 3后 2h即明显下调 ,1 0mg/L ,2 0mg/L ,30mg/L及 40mg/L组分别比H2 O2 组低了 68 71 %、71 0 0 %、86 72 %、95 1 6 %。结论 :丹参单体IH764 - 3可以诱导H2 O2 刺激的HSCs凋亡 ,下调FAKmRNA表达是其诱导HSCs凋亡的机制之一。AIM: To investigate the effect of IH764-3 on the apoptosis of H 2O 2-stimulated hepatic stellate cells(HSCs) and the alteration of focal adhesion kinase (FAK). METHODS: The proliferation of HSCs was examined by direct cell count and the apoptosis was determined by Annexin-V/PI labeling, while the morphological change was observed by light microscopy and transmission electron microscopy. In addition, FAK mRNA was detected by RT-PCR. RESULTS: H 2O 2 promoted the proliferation of HSCs. IH764-3 induced the apoptosis of HSCs in a dose-dependent manner. The HSC apoptotic rates of different groups were 6.35%,9.28%,15.10%,19.69%,respectively, after treated with different concentrations of IH764-3 for 48 h while H 2O 2 group showed 2.30%. In 30 mg/L group, the apoptosis rates were 6.73%?10.34%?15.10% for the indicated time periods(12 h, 24 h, 48 h). In the presence of IH764-3, FAK mRNA decreased. The FAK mRNA reduction began at 2 h after adding IH764-3. CONCLUSION: IH764-3 induced the apoptosis of HSCs. Down-regulating the expression of FAK mRNA may be one of its mechanisms.

关 键 词：丹参单体 肝星状细胞 细胞凋亡 粘着斑激酶 过氧化氢 

分 类 号：R285.5[医药卫生—中药学]