阳离子脂质体转染Akt基因对缺血性心脏病细胞凋亡的影响

Effect of transgenic human Akt gene ion apoptosis of cardiac myocyte after ischemia-reperfusion injury

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 explore the effect and mechanism of transgenic human Akt gene against ischemia-reperfusion （I-R） apoptosis in cardiac myocytes in vitro. Methods Cardiac myocytes from new born SD rats were isolated and cultured in vitro, which were randomly divided into five groups of blank group（A）, I-R group（B）, Akt group（C）, vehicle blank group（D） and Akt blockade group（E）. The levels of lactate dehydrogenase（LDH） and the cell activity in the medium were tested and cell apoptosis was evaluated by flow cytometry. The expression levels of Akt, Bcl-2, NF-κB p65 proteins were measured by Western blot. Results Compared with groups of B, D and E;, the LDH was decreased, cell activity was increased, the apoptosis rate was decreased and expression levels of Akt, Bcl-2,NF-κBp65 were increased significantly in group C （P〈0.05）. Conclusion Transgenic plasmid Akt gene shows a specific protective effect on the cultured cardiac myocytes against I-R injury by inhibiting cell apoptosis.