FAD通过激活SCAD抑制tBHP诱导的大鼠H9C2心肌细胞凋亡

FAD inhibits tBHP-induced apoptosis of rat H9C2 cardiomyocytes by activating SCAD

目的:探讨黄素腺嘌呤二核苷酸(FAD)对叔丁基过氧化氢(tBHP)诱导的大鼠心肌细胞凋亡的影响及其机制。方法:体外培养大鼠H9C2心肌细胞,设立对照(Con)组、tBHP组、FAD组及tBHP+FAD组。采用FAD孵育H9C2细胞24 h,tBHP损伤6 h。CCK-8法检测细胞活力;Western blot检测短链酰基辅酶A脱氢酶(SCAD)蛋白表达;RT-qPCR检测SCAD的mRNA表达水平;检测SCAD酶活性,以及游离脂肪酸、ATP和活性氧(ROS)含量。结果:与Con组相比,tBHP组心肌细胞内SCAD的蛋白和mRNA表达减少,SCAD酶活性下降,ATP含量减少(P<0.01),但游离脂肪酸和ROS含量增加(P<0.01)。与tBHP组相比,tBHP+FAD组SCAD的蛋白和mRNA表达增加,SCAD酶活性增加,ATP含量增加(P<0.01),而游离脂肪酸和ROS含量减少(P<0.01)。结论:FAD可抑制tBHP诱导的大鼠H9C2心肌细胞凋亡;该作用可能通过促进心肌细胞SCAD表达、增强脂肪酸β氧化、减少氧化应激、改善能量代谢而实现。

AIM:To explore the effect of flavin adenine dinucleotide(FAD)on H9C2 cardiomyocyte apoptosis induced by tert-butyl hydroperoxide(tBHP)and its mechanism.METHODS:Rat H9C2 cardiomyocytes were divided into four groups:control(Con)group,tBHP group,FAD group and tBHP+FAD group.The H9C2 cells were cultured with FAD for 24 h and then treated with tBHP for 6 h.The cell viability was detected by CCK-8 assay.The protein level of short-chain acyl-CoA dehydrogenase(SCAD)was detected by Western blot.The mRNA level of SCAD was detected by RT-qPCR.The enzyme activity of SCAD,and the content of free fatty acids,ATP and reactive oxygen species(ROS)were also detected.RESULTS:Compared with Con group,the mRNA and protein expression levels of SCAD,the enzyme activity of SCAD and the content of ATP were decreased(P<0.01),while the content of free fatty acids and ROS was increased in tBHP group(P<0.01).Compared with tBHP group,the mRNA and protein expression levels of SCAD,the enzyme activity of SCAD and the content of ATP were increased(P<0.01),while the content of free fatty acids and ROS was decreased in tBHP+FAD group(P<0.01).CONCLUSION:FAD may inhibit H9C2 cardiomyocyte apoptosis by promoting the expression of SCAD,enhancing cardiomyocyte fatty acidβoxidation,reducing oxidative stress and improving energy metabolism.