摘要
目的探讨Cyp2j3基因通过STAT3信号通路对H9C2心肌细胞增殖凋亡的影响及机制。方法 H9C2大鼠心肌细胞分为正常对照组(对照组,常氧条件下培养)、pc DNA3.1组(转染空载体pcDNA3.1)、单纯缺氧复氧组(I/R+pcDNA3.1组,转染pc DNA3.1后,缺氧10 h后复氧2 h)和单纯缺氧复氧+pc DNA3.1-Cyp2j3组转染Cyp2j3的过表达载体后,缺氧10 h后复氧2 h)。Western bloting检测过表达Cyp2j3的效果;噻唑蓝(MTT)法检测各组细胞活力;乳酸脱氢酶(LDH)试剂盒检测各组细胞的LDH活性;流式细胞术检测各组细胞的凋亡率;Western bloting检测增殖相关蛋白Ki67、凋亡相关蛋白含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、B细胞淋巴瘤/白血病-2(Bcl-2)及信号转导与转录因子3(STAT3)信号通路STAT3和磷酸化的STAT3的蛋白表达。结果转染pc DNA3.1-Cyp2j3后的H9C2细胞Cyp2j3的蛋白表达显著升高;I/R+pc DNA3.1组和I/R+pcDNA3.1-Cyp2j3组细胞的OD值及Ki67、Bcl-2和pSTAT3的蛋白表达均显著低于pcDNA3.1组,LDH活性、细胞凋亡率及Caspase3蛋白表达均显著高于pcDNA3.1组(P<0.05),而I/R+pc DNA3.1-Cyp2j3组细胞的OD值及Ki67、Bcl-2和p-STAT3的蛋白表达均显著高于I/R+pc DNA3.1组,LDH活性、细胞凋亡率及Caspase3蛋白表达均显著低于I/R+pcDNA3.1组(P<0.05)。结论过表达Cyp2j3基因可通过激活STAT3信号减弱缺氧复氧引起的H9C2细胞增殖降低、LDH活性和凋亡增加,从而对心肌细胞起保护作用。
Objective To investigate the effect and mechanism of Cyp2j3 gene on proliferation and apoptosis of H9C2 cardiomyocytes through STAT3 signaling pathway. Methods Rats H9 C2 cardiomyocytes cells rats were divided into normal control group（ control group,cells are cultured under the condition of normal oxygen）,pcDNA3. 1（ Cells were transfected into empty carrier pcDNA3. 1）,hypoxia reoxygenation ＋pcDNA3. 1 group（ I/R ＋ pcDNA3. 1 group,after cells were transfected to pcDNA3. 1,reoxygenation 2 h after hypoxia 10 h） and hypoxia reoxygenation ＋ pcDNA3. 1-Cyp2j3 group（ after cells were transfected to pcDNA3. 1-Cyp2j3,reoxygenation 2 h after hypoxia 10 h）,the effect of Cyp2j3 expression was detected by Western bloting; The cell viability in each group were detected by MTT; LDH activity were detected by LDH detection kit; the apoptosis rate were detected by flow cytometry; the expression of Ki67,Caspase3,Bcl-2,STAT3 and p-STAT3 protein were detected by Western bloting. Results The protein expression of Cyp2j3 in H9 C2 cells increased significantly after transfection with pcDNA3. 1-Cyp2j3. The OD value,Ki67,Bcl-2 and p-STAT3 protein expression in I/R ＋ pcDNA3. 1 and I/R ＋ pcDNA3. 1-Cyp2j3 group were significantly lower than pcDNA3. 1 group,LDH activity,cell apoptosis rate and Caspase3 protein expression were significantly higher the pcDNA3. 1 group（ P〈0. 05）. The OD value,Ki67,Bcl-2 and p-STAT3 protein expression in I/R ＋ pcDNA3. 1-Cyp2j3 group were significantly higher than I/R ＋ pcDNA3. 1 group,LDH activity,cell apoptosis rate and Caspase3 protein expression were significantly lower than I/R ＋ pcDNA3. 1 group（ P〈0. 05）. Conclusion Overexpression of Cyp2j3 gene can reduce the proliferation of H9 C2 cells,increase the activity of LDH and increase apoptosis by activating the STAT3 signal,thus protecting cardiomyocytes.
作者
周咏梅
贺涛
ZHOU Yong - mei;HE Tao(Department of Cardiology,Sichuan Provincial People＇s Hospital,Chengdu Sichuan 610000,China.)
出处
《临床和实验医学杂志》
2018年第16期1691-1695,共5页
Journal of Clinical and Experimental Medicine
