大鼠心肌细胞PTEN基因的表达情况及沉默对其氧糖剥夺/复氧模型耐受能力的影响

The expression of PTEN in rat myocardial cells and cell protective effect of silencing of PTEN gene on oxygen-glucose deprivation/reoxygenation model of MC

目的研究第10号染色体同源丢失性磷酸酶张力蛋白基因(PTEN)在大鼠心肌细胞的表达情况,并观察PTEN基因核糖核酸(RNA)干扰沉默后对大鼠心肌细胞氧糖剥夺/复氧模型耐受能力的影响。为临床心肌梗死的治疗提供新思路。方法体外分离培养大鼠心肌细胞并鉴定,使用逆转录-聚合酶链反应(RT-PCR)法检测其PTEN基因表达;使用脂质体法把PTEN基因特异的小干扰RNA(siRNA)转染沉默PTEN基因;制备体外氧糖剥夺/复氧模型,观察PTEN基因的沉默对细胞的保护作用。结果成功培养大鼠心肌细胞并鉴定。RT-PCR检测显示大鼠心肌细胞高表达PTEN基因,通过siRNA成功干扰了大鼠心肌细胞PTEN基因的表达,流式细胞仪检测结果显示PTEN基因沉默后的细胞对氧糖剥夺/复氧模型有较强的耐受能力。结论 PTEN基因沉默后,大鼠心肌细胞耐受氧糖剥夺/复氧环境的能力增强,具有更强的存活能力,为临床心肌梗死区域的心肌细胞存活提供新的治疗思路。

Objective To study the expression of phosphatase and tensin homolog deleted on chromosome 10(PTEN)in rat myocardial cells,and observe the cell protective effect of silencing of PTEN gene on oxygen-glucose deprivation/reoxygenation model of MC,so as to offer better way to cure myocardial infarction.Methods The rat myocardial cells was isolate in vitro and cultured.Reverse transcription-polymerase chain reaction(RT-PCR)method was used to detect the expression of PTEN gene,and liposome method was used to transfect PTEN gene-specific small interfering RNA(siRNA)to silence PTEN gene.An in vitro oxygen-glucose deprivation/reoxygenation model was prepared to observe the protective effect of PTEN gene silencing on cells.Results The rat myocardial cells was successfully cultured and identified.RT-PCR detection showed high expression of PTEN gene in rat myocardial cells,and siRNA successfully interfered with the expression of PTEN gene in rat myocardial cells.The results of flow cytometry showed that the PTEN gene silenced cells had a strong protective effect to the oxygen-glucose deprivation/reoxygenation model.Conclusion After silencing of PTEN gene,the protective effect of rat myocardial cells to oxygen-glucose deprivation/reoxygenation environment is enhanced,and they have stronger survival ability,which provide new treatment ideas for the survival of myocardial cells in clinical myocardial infarction areas.