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目的:研究肿瘤坏死因子-α(TNF-α)对高糖高胰岛素状态下血管平滑肌细胞(VSMC)增殖的影响及其可能机制。方法:采用正常和高糖高胰岛素两种培养基培养兔胸主动脉平滑肌细胞,用四唑盐比色法(MTT法)测定细胞增殖,Griess试剂法测定细胞培养液中NO含量,一氧化氮合酶(NOS)试剂盒法测定NOS活性。结果:在两种培养条件下L-NAME均能促进VSMC增殖。在正常培养条件下左旋精氮酸(L-Arg)、TNF-α不影响VSMC增殖,在高糖高胰岛素培养条件下L-Arg能抑制VSMC增殖,而TNF-α促进VSMC增殖。在高糖高胰岛素培养条件下VSMC的NOS活性及NO产量均低于常规培养,TNF-α增加NOS活性及NO产量。其作用可被L-NAME抑制。TNF-α对L- Arg及L-NAME的作用具有双向调节性。结论:TNF-α能加强L-Arg抑制VSMC增殖的作用,同时也协同L-NAME促进VSMC的增殖,TNF-α诱导VMSC增殖的作用可能与NO介导有关。
Objective: To investigate the effect of tumor necrosis factor-α (TNF-α) on the proliferation of vascular smooth muscle cells (VSMC) under high glucose and high insulin conditions and its possible mechanism. Methods: Smooth muscle cells of rabbit aorta were cultured with normal and high glucose and high insulin. Cell proliferation was measured by tetrazolium salt (MTT) method. The content of NO in cell culture medium was determined by Griess reagent method. Nitric oxide The activity of NOS was measured by the method of NOS kit. RESULTS: L-NAME promoted VSMC proliferation in both culture conditions. L-Arginine (L-Arg) and TNF-α did not affect the proliferation of VSMC under normal culture conditions. L-Arg inhibited the proliferation of VSMC in high glucose and high insulin medium, while TNF-α promoted the proliferation of VSMC. Under high glucose and high insulin culture conditions, NOS activity and NO production of VSMC were lower than that of conventional culture, and TNF-α increased NOS activity and NO production. Its effect can be inhibited by L-NAME. The effect of TNF-α on L-Arg and L-NAME is bidirectional. CONCLUSION: TNF-α can enhance the effect of L-Arg on proliferation of VSMCs, and also promote the proliferation of VSMCs in coordination with L-NAME. The effect of TNF-α on the proliferation of VMSCs may be related to the mediation of NO.