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目的:研究小豆蔻明(CAR)对人脐动脉平滑肌细胞(HUASMCs)增殖的影响,探讨其可能作用机制。方法:采用高糖高胰岛素培养基模拟胰岛素抵抗(IR)诱导HUASMCs增殖,MTT法考察CAR对细胞增殖的作用,荧光实时定量PCR检测哺乳类雷帕霉素靶蛋白(mTOR)及其结合蛋白Raptor,Rictor mRNA表达,Livak法计算mRNA表达量。结果:高糖高胰岛素培养基明显促进HUASMCs的增殖。低剂量CAR(1×10-8,1×10-7,1×10-6mol.L-1)对正常及模拟IR培养的HUASMCs增殖无影响,而高剂量CAR(1×10-5,1×10-4mol.L-1)抑制细胞增殖。模拟胰岛素抵抗状态下,PD98059和LY294002干预后,CAR(1×10-5mol.L-1)对磷脂酸(PA)诱导的增殖具有抑制作用,可降低mTOR mRNA的表达,同时减少Raptor,Rictorm RNA的含量。结论:CAR抑制细胞增殖机制与mTOR通路密切相关,可能直接或主要作用于mTOR。
Objective: To investigate the effect of cardamine (CAR) on the proliferation of human umbilical artery smooth muscle cells (HUASMCs) and to explore its possible mechanism. Methods: High glucose and high insulin medium was used to simulate insulin resistance (IR) -induced proliferation of HUASMCs. MTT assay was used to detect the effect of CAR on cell proliferation. Real-time quantitative PCR was used to detect mammalian target of rapamycin (mTOR) and its binding protein Raptor , Rictor mRNA expression, Livak method to calculate the mRNA expression. Results: High glucose and high insulin medium significantly promoted the proliferation of HUASMCs. Low dose CAR (1 × 10-8, 1 × 10-7, 1 × 10-6mol.L-1) had no effect on proliferation of HUASMCs cultured in normal and simulated IR, while high dose CAR (1 × 10-5,1 × 10-4mol.L-1) inhibited cell proliferation. Under simulated insulin resistance, CAR (1 × 10-5mol.L-1) inhibited the proliferation induced by phosphatidic acid (PA) after the intervention of PD98059 and LY294002, which could reduce the expression of mTOR mRNA and reduce the expression of Raptor and Rictorm RNA Content. CONCLUSION: The mechanism of CAR inhibition of cell proliferation is closely related to mTOR pathway and may directly or mainly affect mTOR.