摘要
目的研究前蛋白转化酶枯草溶菌素9(PCSK9)基因沉默后对血管平滑肌细胞(VSMC)增殖的影响,初步探讨低密度脂蛋白受体相关蛋白1(LRP-1)蛋白表达在其中的作用。方法建立PCSK9小分子干扰RNA(siRNA)模型,培养VSMC,分为空白对照组、阴性转染组和阳性转染组,选取转染效率最高的PCSK9siRNA对VSMC进行干预,分别于12、24、36、48、60h用MTT法测定吸光度(A)值,绘制增殖曲线,观察其对VSMC增殖的影响,转染48h后用Western blot印迹法检测细胞内LRP-1蛋白表达情况。结果与阴性转染组比较,阳性转染组24、36、48和60h的A值明显下降(P<0.05),PCSK9siRNA抑制VSMC的增殖(P<0.05)。与空白对照组和阴性转染组比较,阳性转染组LRP-1蛋白表达明显上调(P<0.05)。结论 PCSK9siRNA能有效抑制VSMC的增殖作用,且这种作用可能与LRP-1蛋白表达上调有一定的关系。
Objective To study the effect of PCSK9 siRNA on proliferation of vascular smooth muscle cells(VSMC)and the role of LRP-1expression in proliferation of VSMC.Methods After a PCSK9 siRNA model was established,the cultured VSMC were divided into blank control group,negative transfection group and positive transfection group.The absorbance value(A)of VSMC was detected by MTT assay.Curves of proliferative VSMC were plotted.Effect of PCSK9 siRNA on proliferation of VSMC was detected at hours 12,24,36,48,60 respectively.LRP-1expression in VSMC was detected by Western blot at 48 hafter transfection.Results The Avalue was significantly lower in positive transfection group than in negative transfection group at hours24,36,48 and 60(P〈0.05).The LRP-1expression level was significantly higher in positive transfection group than in blank control group and negative transfection group(P〈0.05).Conclusion PCSK9 siRNA can effectively inhibit the proliferation of VSMC by up-regulating the LRP-1 expression.
出处
《中华老年心脑血管病杂志》
CAS
2016年第7期734-738,共5页
Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基金
湖南省2014年科技创新项目(2014658)
2012年仁术科研发展基金(201219)