小豆蔻明对人脐动脉平滑肌细胞增殖的影响及机制研究

Antiproliferation of cardamonin associated with mRNA expression of mTOR,Raptor and Rictor

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 antiproliferation effect of cardamonin（CAR） and its possible mechanisms on human umbilical artery smooth muscle cells（HUASMCs） cultured in the mimicking insulin resistance（IR） medium.Method： Proliferation of HUASMCs was assayed by MTT method.The mRNA expression of mTOR,Raptor and Rictor was detected by a real-time PCR.The expression content was calculated by Livak method using internal control of β-actin.Result： The proliferation of HUASMCs cultured in the mimicking IR medium was significantly increased.Both in normal and mimic IR culture medium,cells proliferation was inhibited by CAR（1×10-5,1×10-4 mol·L-1）.Pretreated with PD98059 and LY294002,cell proliferation induced by phosphatidic acid（PA） was inhibited,and the mRNA expression of mTOR,Raptor and Rictor was significantly decreased by CAR in the mimic IR medium.Conclusion： It is implicated that antiproliferation of CAR is involved in mRNA expression decrease of mTOR and its relative protein Raptor and Rictor.