丙泊酚降低高糖环境下H9C2心肌细胞缺氧后的损伤

目的探讨小窝蛋白3(Cav-3)在丙泊酚降低高糖环境下H9C2心肌细胞缺氧后损伤中的作用。方法培养大鼠原代H9C2心肌细胞,构建细胞缺氧-复氧模型。将心肌细胞分为六组:正常培养组(NC组)、高糖培养组(HG组)、高糖缺氧-复氧组(HR组)、丙泊酚处理组(P组)、Cav-3抑制剂组(PI组)和DMSO溶剂组(D组)。比较六组心肌细胞损伤程度、氧化应激反应、线粒体功能、Cav-3蛋白含量、蛋白激酶B(AKT)及信号传导及转录激活因子3(STAT3)活化情况。结果与HR组比较,P组细胞活力、总SOD(T-SOD)活性、线粒体活力、ATP含量、Cav-3蛋白含量、AKT及STAT3活化明显增加,LDH、CK-MB活性、cTnI含量、Bcl-2相关X蛋白(Bax)与B淋巴细胞瘤-2(Bcl-2)比值(Bax/Bcl-2)明显下降,含半胱氨酸的天冬氨酸蛋白水解酶-3(caspase 3)及含半胱氨酸的天冬氨酸蛋白水解酶-9(caspase-9)蛋白含量、MDA浓度、JC-1染色的荧光绿红比明显降低,DCF-DA荧光阳性细胞数、MPTP开放明显减少(P0.05);与P组比较,PI组和D100组细胞活力、T-SOD活性、线粒体活力、ATP含量、Cav-3蛋白含量明显降低,AKT及STAT3活化明显减少(P0.05),LDH、CK-MB活性、cTnI含量、Bax/Bcl-2值、caspase-3及caspase-9蛋白含量、MDA浓度、JC-1染色的荧光绿红比明显升高,DCF-DA荧光阳性细胞数、MPTP开放明显增加(P0.05)。结论丙泊酚通过上调Cav-3减少高糖环境下H9C2心肌细胞的缺氧后线粒体的损伤及细胞的死亡。

Role of propofol of protecting H9C2 myocardial against hypoxia injury under hyperglycemia

Objective To investigate the role of Cav-3 in the protection of cardiomyocytes against hypoxia by propofol.
Methods Rat primary H9C2 cardiomyocytes were cultured to establish cell hypoxia-reoxygenation model. The cells were assigned to the following groups: normal control group (group NC), high glucose group (group HG), HR under high glucose group (group HR), propofol treated group (group P), Cav-3 inhibitor group (group PI) or the solvent DMSO group (group D). The degree of cardiomyocyte injury, myocardial oxidative stress response, mitochondrial function, Cav-3 protein expression, AKT and STAT3 activation were compared.
Results Compared with group HR, the cell viability, total SOD (T-SOD), mitochondrial activity, ATP content, Cav-3 protein content, AKT and STAT3 activation in group P were significantly increased (P < 0.05), while LDH, CK-MB, cTnI, the ratio of Bax to Bcl-2, caspase-3 and caspase-9, the number of DCF-DA positive cells, the fluorescence green-red ratio of MDA and JC-1 staining and MPTP opening were significantly decreased (P < 0.05). Compared with group P, cell viability, total SOD (T-SOD), mitochondrial activity, ATP content, Cav-3 protein content, AKT and STAT3 activation in group PI and group D100 were significantly decreased, while LDH, CK-MB, cTnI, the ratio of Bax to Bcl-2, caspase-3 and caspase-9, the number of DCF-DA positive cells, the fluorescence green-red ratio of MDA and JC-1 staining and MPTP opening were significantly increased (P < 0.05).
Conclusion Propofol attenuates mitochondrial damage and cell death after hypoxia in H9C2 cardiomyocytes by up-regulating Cav-3.