摘要
目的:差异筛选淫羊藿苷(icariin,ICA)诱导血管平滑肌细胞(vascular smooth muscle cell,VSMC)凋亡相关基因葡萄糖调节蛋白78(glucose regulated protein78,GRP78),观察其在动脉粥样硬化(athemsclerosis,AS)病灶中的表达情况,探讨ICA抗AS的可能机制。方法:建立同型半胱氨酸(homocysteinemia,HCY)诱导的兔VSMC增殖模型,与不同浓度ICA共同培养48h后,原位缺口末端标记(TUNEL)法检测细胞凋亡率;逆转录聚合酶链式反应(RT-PCR)筛选差异基因片段,克隆、鉴定、测序,组织原位杂交观察该基因在正常及AS病灶中的表达水平。结果:TUNEL法显示ICA组凋亡细胞数目明显增多;筛选出全长648bp的基因片断与兔GRP78基因高度同源,组织原位杂交显示该基因在AS病灶VSMC胞质中高表达。结论:GRP78基因参与AS病变形成,其表达上调可能是ICA诱导VSMC凋亡作用靶点之一。
Objective：To select the differentially expressed vascular smooth muscle cell（VSMC） apoptosis gene induced by icariin,glucose regulated protein 78（GRP78）,to observe the expression of GRP78 in atherosclerosis tissue and to the elucidate the mechanism of anti-atherosclerosis of icariin.Methods：The model was established by using halfcystine in viro.The vascular smooth muscle cell was incubated with various concentrations of icariin for 48 hours.The apoptosis was detected by TUNEL assay.GRP78 gene was amplificated by PCR,recombinant plasmid were constructed,transfected into E.coli,positive clonings were selected and conf irmed by restriction endonucleas analysis,and sequence-ed.Tissue situ hybridization was used to observe the GRP78 expression in atherosclerosis tissue.Results：Icariin promoted obviously vascular smooth muscle cell apoptosis.Furthermore,the fragment,648bp,was similar to the gene of GRP78 from VSMC,and the expression level was signif icantly higher in high cholesterol group than in control nroup.Conclusions：The GRP78 plays a key role in inducing atherosclerotic lesions.The mechanism of anti-atherosclerosis of icariin might be partly due to promoting apoptosis of vascular smooth muscle cell.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2010年第5期771-774,共4页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
河南省科技攻关计划项目(No.0624420026)~~
