琥珀酸对原代心肌细胞缺氧复氧损伤的保护作用

本研究采用SD大鼠乳鼠心肌细胞原代培养,复制心肌细胞缺氧复氧损伤模型,考察琥珀酸对心肌细胞缺氧复氧损伤的LDH漏出率的影响,并进一步采用流式细胞术及Western blot考察琥珀酸对心肌细胞凋亡,cleaved caspase-3及p-Akt的影响,探讨琥珀酸对新生大鼠原代心肌细胞缺氧复氧损伤的保护作用。研究结果发现:①琥珀酸31.25~500 mg·L^-1对原代心肌细胞活力无明显影响,琥珀酸400,200,100,50 mg·L^-1均可显著降低心肌细胞缺氧复氧损伤LDH漏出率(P<0.01或P<0.05);②琥珀酸400,200 mg·L^-1可显著降低缺氧复氧损伤所致的心肌细胞凋亡百分数(P<0.05),抑制心肌细胞缺氧复氧所致的cleaved caspase-3 蛋白表达增加(P<0.05);③琥珀酸400 mg·L^-1可显著增加心肌细胞的p-Akt蛋白表达(P<0.05),而琥珀酸200 mg·L^-1对p-Akt蛋白表达无明显影响。因此,本研究认为琥珀酸可通过激活Akt的磷酸化而抑制心肌细胞缺氧复氧所致的坏死和凋亡。

Protective effect of succinic acid on primary cardiomyocyte hypoxia/reoxygenation injury

To establish cardiomyocyte hypoxia/reoxygenation injury model by culturing primary cardiomyocytes from suckling SD rats, in order to study the effect of succinic acid on LDH leakage rate cardiomyocyte ischemia/reperfusion injury. Furthermore, flow cytometry and western blot were conducted to detect the effect of succinic acid on cardiomyocyte apoptosis, cleaved caspase-3 and p-Akt, and discuss the protective effect of succinic acid on primary cardiomyocyte hypoxia/reoxygenation injury of primary cardiomyocytes from neonatal SD rats. According to the findings of the study, succinic acid at the concentrations ranging between 31.25 mg·L^-1 and 500 mg·L^-1 had no significant effect on primary cardiomyocyte activity, and succinic acid at the concentrations of 400, 200, 100, 50 mg·L^-1 could notably reduce cardiomyocyte ischemia/reperfusion LDH leakage rate (P<0.01 or P<0.05, respectively). Succinic acid at the concentrations of 400 mg·L^-1 and 200 mg·L^-1 could significantly reduce the percentage of cardiomyocyte apoptosis (P<0.05), and inhibit the protein expression of cleaved caspase-3 caused by cardiomyocyte ischemia/reperfusion (P<0.05). Succinic acid at the concentration of 400 mg·L^-1 could remarkably increase the protein expression of cardiomyocyte Akt (P<0.05), while succinic acid at the concentration of 200 mg·L^-1 had no obvious effect on the protein expression of cardiomyocyte Akt. Therefore, this study demonstrated that succinic acid could inhibit necrosis and apoptosis caused by cardiomyocyte hypoxia/reoxygenation by activating Akt phosphorylation.