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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 investigate the effect of evodiamine on the proliferation of rat vascular smooth muscle cells (VSMCs) induced by angiotensin Ⅱ (Ang Ⅱ) and its mechanism. Methods VSMCs of rat thoracic aorta were cultured by tissue block adhesion method. The proliferation of VSMCs was detected by MTT assay. The cell cycle was detected by flow cytometry. The activity of nitric oxide synthase (NOS) and the content of nitric oxide (NO) in the culture supernatant were measured by real time RT-PCR. The expression of proliferating cell nuclear antigen (PCNA), endothelial NOS Expression to explore the possible mechanism of Evo. Results 1μmol·L-1AngⅡignificantly increased the OD of VSMCs, increased the S phase in the cell cycle and decreased the ratio of G0 / G1 phase cells, significantly reduced the activity of NOS and the content of NO in the supernatant of cultured cells, NOS increased the expression of PCNA mRNA; 0.01, 0.1, 1μmol·L-1 evodiamine inhibited the effect of AngⅡ in a concentration-dependent manner, and decreased the S phase in the cell cycle and increased the ratio of G0 / G1 phase cells. Conclusion Evodiamine can promote the synthesis and release of NO. One of the mechanisms that inhibits the proliferation of VSMCs through arresting VSMCs in G0 / G1 phase is evodiamine.