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目的研究转染基因Akt对心肌细胞凋亡的影响。方法培养新生SD大鼠心肌细胞;提取人Akt质粒,用脂质体将其转染心肌细胞。实验分5组:空白对照(A)组、缺血-再灌注损伤对照(B)组、Akt基因(C)组、阳离子脂质体空载体(D)组和Akt抑制剂(E)组;各组进行模拟缺血-再灌注后测定乳酸脱氢酶(LDH)含量、细胞活性、细胞凋亡率及Akt、Bcl-2、NF-κB p65蛋白表达。结果C组LDH含量较B组、D组和E组明显减少(P<0.05),细胞活性高(P<0.05),细胞凋亡率低(P<0.05),Akt、Bcl-2与NF-κB p65蛋白表达增加(P<0.05)。结论细胞凋亡参与了心肌缺血-再灌注损伤过程,转染Akt基因可减少缺血-再灌注损伤心肌细胞凋亡率。
Objective To study the effect of transfection gene Akt on cardiomyocyte apoptosis. Methods Cardiomyocytes of neonatal SD rats were cultured. Human Akt plasmid was extracted and transfected into cardiomyocytes with liposome. The experiment was divided into five groups: blank control group (A), ischemia - reperfusion injury control group (B), Akt gene group (C), cationic liposome empty vector group (D) and Akt inhibitor group The levels of lactate dehydrogenase (LDH), cell viability, apoptotic rate and expression of Akt, Bcl-2, NF-κB p65 protein were measured in each group after simulated ischemia-reperfusion. Results The level of LDH in group C was significantly lower than that in group B, D and E (P <0.05), the cell activity was higher (P <0.05) κB p65 protein expression increased (P <0.05). Conclusions Apoptosis is involved in the process of myocardial ischemia - reperfusion injury. Transfection of Akt gene can reduce the rate of myocardial cell apoptosis induced by ischemia - reperfusion.