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目的:探讨拉马克啉(levcromakalim)对内皮素-1(ET-1)诱导的大鼠血管平滑肌细胞增殖及蛋白激酶C(protein kinase C,PKC)表达的影响。方法:体外培养Wistar大鼠主动脉血管平滑肌细胞(vascularsmooth muscle cell,VSMCs),用ET-1诱导其增殖,用不同浓度拉马克啉共培养,MTT法评价VSMCs增殖情况,3H-TdR检测DNA合成,流式细胞术检测VSMCs凋亡,逆转录聚合酶链式反应(RT-PCR),Western blot检测PKCαmRNA及蛋白表达。结果:拉马克啉对ET-1所致VSMCs增殖有显著抑制作用,随浓度增加,其MTT活性细胞含量和3H-TdR掺入量都明显减少(P<0.05);拉马克啉呈剂量依赖性地增加G0/G1期VSMCs(P<0.05),促使VSMCs凋亡增多(P<0.05);拉马克啉抑制VSMCs内PKCαmRNA及蛋白表达。结论:拉马克啉抑制ET-1诱导的VSMCs增殖作用,可能的机制是通过下调VSMCs内PKCα表达水平而影响VSMCs增殖和凋亡。
Objective: To investigate the effects of levcromakalim on the proliferation and the expression of protein kinase C (PKC) in rat vascular smooth muscle cells induced by endothelin-1 (ET-1). Methods: Vascularsmooth muscle cells (VSMCs) of Wistar rats were cultured in vitro. The proliferation of VSMCs was induced by ET-1. The proliferation of VSMCs was evaluated by MTT assay. DNA synthesis was detected by 3H-TdR The apoptosis of VSMCs was detected by flow cytometry. The mRNA and protein expressions of PKCα were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. RESULTS: Ramipril had a significant inhibitory effect on the proliferation of VSMCs induced by ET-1, and the MTT activity and 3H-TdR incorporation were significantly decreased with increasing concentration (P <0.05). There was a dose-dependent (P <0.05), and increased the apoptosis of VSMCs (P <0.05). And the inhibition of PKCαmRNA and protein expression in VSMCs by lamacline. CONCLUSION: The inhibitory effect of Ramamarine on the proliferation of VSMCs induced by ET-1 may be related to the inhibition of proliferation and apoptosis of VSMCs by down-regulating the expression of PKCα in VSMCs.