Roscovitine对TNF-α诱导的大鼠血管平滑肌细胞增殖及细胞周期的影响  

Effects of roscovitine on proliferation and cell cycle induced by TNF-α of vascular smooth muscle cells in rats

作　　者：王超[1] 刘玉 安然 赵京山 孙佳欢 李爱英 WANG Chao;LIU Yu;AN Ran;ZHAO Jing-shan;SUN Jia-huan;LI Ai-ying(Key Lab of Geriatrics of Hebei General Hospital,Shijiazhuang050051,China;Dept of Biochemistry and Biology,Basic Medical College,Hebei University of Chinese Medicine and Hebei Key Lab of Chinese Medicine Research on Cardiocerebrovascular Diseases,Shijiazhuang050200,China)

机构地区：[1]河北省人民医院代谢病重点实验室,河北石家庄050051 [2]河北中医学院基础医学院生物化学与生物学教研室河北省心脑血管中医药防治重点实验室,河北石家庄050200

出　　处：《中国药理学通报》2019年第3期376-381,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 31271466;81573698);河北省自然科学基金资助项目(No H2017423033);河北省教育厅项目(No ZD2014005;ZD2017056)

摘　　要：目的观察Roscovitine对TNF-α诱导的大鼠血管平滑肌细胞(VSMC)增殖及细胞周期的影响。方法组织贴块法培养大鼠VSMC细胞,采用TNF-α诱导其增殖,加入不同浓度的Roscovitine预处理15 h,将细胞分为:对照组、TNF-α组、Roscovitine 5、10、15、30μmol·L^(-1)组。MTT比色法检测细胞增殖活性;Western blot检测增殖细胞核抗原(PCNA);流式细胞仪检测细胞周期;荧光定量RT-PCR及Western blot检测细胞周期蛋白(Cyclin A、Cyclin B、Cyclin D、Cyclin E)、细胞周期蛋白依赖激酶(CDK4、CDK5)、细胞周期抑制蛋白(p53、p21、p27)的表达。结果 Roscovitine能抑制VSMC增殖;抑制细胞周期从G_0/G_1期向S期转化。与TNF-α组比较,Roscovitine 5、10、15、30μmol·L^(-1)组能降低细胞周期蛋白Cyclin A、Cyclin B、Cyclin D、Cyclin E蛋白表达,降低细胞周期蛋白依赖激酶CDK4、CDK5蛋白表达,升高细胞周期抑制蛋白p53、p21、p27蛋白表达(P<0.05)。结论 Roscovitine可抑制大鼠VSMC细胞周期进程及增殖活性。Aim To observe the effect of roscovitine on TNF-α-induced proliferation and cell cycle of rat vascular smooth muscle cells(VSMCs).Methods VSMCs were isolated and cultured using tissue explant method.Tumor necrosis factor-alpha(TNF-α)was used to induce the proliferation of VSMCs.VSMCs were divided into control group,TNF-αgroup and roscovitine 5,10,15,30μmol·L^-1 group.Cell proliferation was detected with MTT,and PCNA expression was tested.Flow cytometry was used to detect the cell cycle.The expressions of cyclin proteins(Cyclin A,Cyclin B,Cyclin D,Cyclin E),cyclin-dependent kinase proteins(CDK4,CDK5),and cell cycle inhibitory proteins(p53,p21,p27)were examined by real-time reverse transcription PCR(qRT-PCR)and Western blot.Results Roscovitine could inhibit the proliferation of VSMCs.Roscovitine inhibited transformation of cell cycle from G0/G1 phase to S phase.Compared with TNF-αgroup,the expressions of Cyclin A,Cyclin B,Cyclin D,Cyclin E protein decreased,the expressions of CDK4 and CDK5 protein decreased,while the expression of p53,p21 and p27 protein in cell cycle increased of roscovitine 5,10,15,30μmol·L-1 group(P<0.05).Conclusion Roscovitine inhibits the cell cycle progression and proliferative activity of rat VSMCs.

关 键 词：ROSCOVITINE 血管平滑肌细胞 增殖 细胞周期 细胞周期蛋白 细胞周期蛋白依赖性激酶 细胞周期抑制蛋白 

分 类 号：R-332[医药卫生] R322.12