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目的探讨L-精氨酸(L-Arg)对高糖高胰岛素(High glucose and insulin,HGI)诱导心肌细胞肥大的抑制作用。方法体外培养乳鼠心肌细胞,将细胞分为正常对照组(5.5mmol/L葡萄糖)、模型组(HGI组,给予25.5mmol/L葡萄糖与0.1μmol/L胰岛素)、L-Arg低、中、高剂量组(给予HGI组药物,并分别加入0.01、0.1和1.0mmol/L的L-Arg)和L-Arg+L-NAME组[给予HGI组药物,并加入0.1mmol/LL-Arg和一氧化氮合酶(Nitric oxide synthase,NOS)特异性抑制剂L-NAME],以细胞表面积、蛋白质含量和心房利钠因子(Atrial natriuretic factor,ANF)mRNA表达为反映心肌肥大的指标,观察L-Arg对HGI致心肌细胞肥大作用的影响;采用Real-timePCR法检测内皮型一氧化氮合酶(Endothelial NOS,eNOS)和诱生型一氧化氮合酶(Inducible NOS,iNOS)mRNA的表达;比色法和硝酸还原法分别检测细胞培养液中NOS的活性和NO的含量。结果 L-Arg呈浓度依赖性地抑制HGI诱导的心肌细胞肥大;并上调eNOS mRNA的表达及NOS的活性,增加NO的浓度。NOS抑制剂L-NAME可完全阻断L-Arg的上述作用。结论 L-Arg可通过激活eNOS表达,促进NO释放,产生抗HGI诱导心肌肥大的作用。
Objective To investigate the inhibitory effect of L-arginine on the hypertrophy of cardiomyocytes induced by high glucose and insulin (HGI). Methods The neonatal rat cardiomyocytes were cultured in vitro and divided into normal control group (5.5 mmol / L glucose), model group (HGI group, 25.5 mmol / L glucose and 0.1 μmol / L insulin) High dose group (given HGI group and added 0.01, 0.1 and 1.0 mmol / L L-Arg respectively) and L-Arg + L-NAME group [given HGI group with 0.1 mmol / L L-Arg and Nitric oxide synthase (NOS) inhibitor L-NAME was used to measure the cell surface area, protein content and ANF mRNA expression, Arg on HGI-induced cardiomyocyte hypertrophy; Real-time PCR was used to detect the expression of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) mRNA; The colorimetric method and nitrate reduction method were used to detect the activity of NOS and NO content in cell culture solution respectively. Results L-Arg inhibited HGI-induced cardiomyocyte hypertrophy in a concentration-dependent manner and up-regulated eNOS mRNA expression and NOS activity and increased NO concentration. The NOS inhibitor L-NAME completely blocked the above effects of L-Arg. Conclusion L-Arg can promote the release of NO by activating the expression of eNOS and induce the anti-HGI-induced cardiac hypertrophy.