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目的探讨α-硫辛酸对高糖诱导的乳鼠心肌细胞肥大保护作用及其机制。方法取出生1～3 d乳鼠心尖部心肌细胞原代培养,用低糖培养基培养48 h后分为对照组、高糖组(25.5 mmol/L)、α-硫辛酸组(高糖+50、100、200、300 mg/L)、蛋白激酶C(PKC)抑制剂组(高糖+PKC抑制剂50 nmol/L)、核转录因子-κB(NF-κB)抑制剂组(高糖+NF-κB抑制剂5 mmol/L);细胞培养48 h,检测细胞表面积以及蛋白含量,Wesrtern blot法检测PKC-α、NF-κB、肿瘤坏死因子-α(TNF-α)、立早基因c-fos蛋白表达。结果与对照组比较,高糖组乳鼠心肌细胞肥大、细胞表面积增大、蛋白含量增多,心肌细胞内PKC-α(0.89±0.03)、NF-κB(0.88±0.14)、c-fos(0.62±0.14)、TNF-α(0.85±0.05)蛋白表达均升高(P<0.05);α-硫辛酸能够减轻乳鼠心肌细胞肥大,使心肌细胞表面积减小、蛋白含量减少(P<0.05),与高糖组比较,α-硫辛酸300 mg/L组细胞内PKC-α(0.44±0.07)、NF-κB(0.24±0.03)、TNF-α(0.39±0.05)、c-fos(0.15±0.06)蛋白表达均下降(P<0.05)。结论α-硫辛酸可抑制高糖诱导的乳鼠心肌细胞肥大,其机制可能与抑制PKC/NF-κB/TNF-α/c-fos的信号转导通路有关。
Objective To investigate the protective effect and its mechanism of α-lipoic acid on hypertrophy induced by high glucose in neonatal rat cardiomyocytes. Methods Primary cultured cardiomyocytes of neonatal rats were devided into control group, high glucose group (25.5 mmol / L), α-lipoic acid group (high glucose +50 , 100, 200 and 300 mg / L), PKC inhibitor group (high glucose + 50 nmol / L inhibitor of PKC) and NF-κB inhibitor group NF-κB inhibitor 5 mmol / L). The cells were cultured for 48 h, the cell surface area and protein content were detected. The protein expression of PKC-α, NF-κB, TNF- -fos protein expression. Results Compared with the control group, the cardiomyocyte hypertrophy, the cell surface area and the protein content were increased in the high glucose group. The levels of PKC-α (0.89 ± 0.03), NF-κB (0.88 ± 0.14) and c-fos (P <0.05); α-lipoic acid can reduce myocardial cell hypertrophy, myocardial cell surface area decreased, protein content decreased (P <0.05) (0.44 ± 0.07), NF-κB (0.24 ± 0.03), TNF-α (0.39 ± 0.05) and c-fos (0.15) in the a-lipoic acid 300 mg / ± 0.06) protein expression decreased (P <0.05). CONCLUSION: Alpha-lipoic acid can inhibit the hypertrophy induced by high glucose in neonatal rat cardiomyocytes. The mechanism may be related to the inhibition of signal transduction pathway of PKC / NF-|ÊB / TNF-|Á / c-fos.