吴茱萸碱抑制血管紧张素Ⅱ致大鼠血管平滑肌细胞增殖的研究  

Effect of evodiamine on the vascular smooth muscle cells proliferation induced by Ang Ⅱ in rats

作　　者：侯化化[1,2] 张婧怡[1] 林淑娴[1] 徐洋[1] 李强[3] 孙安盛[1] 

机构地区：[1]遵义医学院药理学教研室贵州省基础药理学重点实验室,贵州遵义563099 [2]贵州航天医院,贵州遵义563000 [3]遵义市第一人民医院,贵州遵义563002

出　　处：《华西药学杂志》2014年第6期645-647,共3页West China Journal of Pharmaceutical Sciences

基　　金：国家自然科学基金资助项目(批准号:81160528)

摘　　要：目的探讨吴茱萸碱对血管紧张素Ⅱ(AngⅡ)所致大鼠血管平滑肌细胞(VSMCs)增殖周期的影响,并探讨其机制。方法利用组织块贴壁法培养大鼠胸主动脉VSMCs,通过MTT比色法检测VSMCs的增殖,采用流式细胞仪检测细胞周期。通过测定培养细胞上清液中一氧化氮合酶(NOS)的活性和一氧化氮(NO)的含量、Real time RT-PCR检测增殖细胞核抗原(PCNA)、内皮型NOS(e NOS)mRNA的表达来探讨Evo可能的作用机制。结果 1μmol·L-1AngⅡ能明显增加VSMCs的OD值,升高细胞周期中S期而降低G0/G1期细胞的比率,明显减少培养细胞上清液中NOS的活性和NO的含量,同时下调e NOS而上调PCNAmRNA的表达;0.01、0.1、1μmol·L-1吴茱萸碱呈浓度依赖性抑制AngⅡ的上述作用,减少细胞周期中S期而升高G0/G1期细胞的比率。结论吴茱萸碱可促进NO的合成、释放,通过阻滞VSMCs于G0/G1期是抑制VSMCs增殖的机制之一。OBJECTIVE To observe the influences of evodiamine on the cell cycle of vascular smooth muscle cells（ VSMCs） proliferation induced by angiotensinⅡ（ AngⅡ） and explore the mechanisms. METHODS VSMCs from rat artery were cultured by tissue explant method and AngⅡ of 1 μmol·L- 1was taken as an inducer. The VSMCs proliferation and cell cycle were respectively measured by the MTT method and flow cytometry. The activity of nitric oxide synthetase（ NOS） and the content of nitric oxide（ NO） in supernatants of VSMCs were detected,respectively. For examining the mechanisms,the mRNA expressions of endothelial nitric oxide synthase（ e NOS） and proliferating cell nuclear antigen（ PCNA） of VSMCs were measured by real time RT- PCR. RESULTS Compared with control group,1 μmol·L- 1AngⅡ significantly increased the absorbance of MTT and the percentage of S and G2/ M phase cells,but G0/ G1 phase cells were decreased in the VSMCs cycle. At the same time,1 μmol·L- 1AngⅡ obviously decreased the activity of NOS and the content of NO in the supernatant of VSMCs,accompanied by up- regulating PCNA mRNA expression and down- regulating e NOS mRNA expression. Evodiamine（ 0. 01- 1 μmol·L- 1） attenuated Ang II- induced VSMCs proliferation and blocked cell transition from G0/ G1 to S phase,increased the e NOS mRNA expression,NOS activity and NO release. CONCLUSION Evodiamine could inhibit the VSMCs proliferation induced by AngⅡ（ 1 μmol·L- 1）,and this effect is partly due to the promotion of NO release and blockage cell transition from G0/ G1 to S phase in the VSMCs cycle.

关 键 词：吴茱萸碱 血管平滑肌细胞 增殖 细胞周期 血管紧张素Ⅱ 一氧化氮 

分 类 号：R96[医药卫生—药理学]