CAJ | 학술논문

目的探讨慢病毒介导的P38丝裂原活化蛋白激酶(P38MAPK)短发夹环RNA(shRNA)对醛固酮过负荷大鼠心肌梗死(心梗)后心功能的影响并探讨其机制. 方法制作醛固酮过负荷大鼠心梗模型,构建慢病毒P38MAPK shRNA(PGLV-shRNA)测序鉴定并经尾静脉注射,超声评价心功能,检测心肌细胞凋亡、P38MAPK mRNA、蛋白及Caspase-3蛋白的表达. 结果假手术组、PGLV空载组和PGLV-SH RNA组细胞凋亡指数分别为(15.20±2.18)％、(31.26±4.45)％和(22.35±3.59)％；醛固酮过负荷大鼠心梗后心脏收缩功能显著降低,伴随心肌细胞凋亡增加、P38MAPK mRNA、蛋白及caspase-3蛋白表达上调(P＜0.01).PGLV-shRNA明显改善心梗后的心脏功能,减少心肌细胞凋亡P38MAPK mRNA、蛋白及caspase-3表达(P＜0.05). 结论醛固酮过负荷大鼠心梗后心脏功能降低与P38MAPK信号通路介导的心肌细胞凋亡相关,PGLV-shRNA抑制细胞凋亡,改善心梗后的心脏功能.
목적 탐토만병독개도적P38사렬원활화단백격매(P38MAPK)단발협배RNA(shRNA)대철고동과부하대서심기경사(심경)후심공능적영향병탐토기궤제. 방법 제작철고동과부하대서심경모형,구건만병독P38MAPK shRNA(PGLV-shRNA)측서감정병경미정맥주사,초성평개심공능,검측심기세포조망、P38MAPK mRNA、단백급Caspase-3단백적표체. 결과 가수술조、PGLV공재조화PGLV-SH RNA조세포조망지수분별위(15.20±2.18)％、(31.26±4.45)％화(22.35±3.59)％；철고동과부하대서심경후심장수축공능현저강저,반수심기세포조망증가、P38MAPK mRNA、단백급caspase-3단백표체상조(P＜0.01).PGLV-shRNA명현개선심경후적심장공능,감소심기세포조망P38MAPK mRNA、단백급caspase-3표체(P＜0.05). 결론 철고동과부하대서심경후심장공능강저여P38MAPK신호통로개도적심기세포조망상관,PGLV-shRNA억제세포조망,개선심경후적심장공능.
Objective To investigate the effects of p38 mitogen-activated protein kinase (MAPK) short hair RNA (shRNA) delivered by lentiviral vectors (pGLV) on cardiac function after myocardial infarction (MI) in aldosterone overload rats and to explore the mechanism.Methods Aldosterone overload rat myocardial infarction model was obtained by ligating the left anterior descending coronary artery.The pGLV-shRNA was constructed,sequenced and injected into rats via tail vein.Rats were divided into 3 groups:pGLV-shRNA group (n=6),pGLV-shRNA-NC group (n=6,contained a nonsense shRNA) and the sham-operation group (n=6).Cardiac function was measured by cardiac ultrasound.Apoptosis was assessed by transferase (TdT)-mediated biotin-16-dUTP nick-end labelling (TUNEL).The p38 MAPK mRNA expression was analyzed by RT-PCR.The protein expressions of p38 MAPK and caspase-3 were detected by Western blot.Results Compared with the sham-operation group,cardiac systolic function was reduced and myocardial apoptosis index was significantly increased [(31.26 ± 4.45) ％ vs.(15.20 ± 2.18) ％,P ＜ 0.01] in pGLV-shRNA-NC group.The mRNA and protien expressions of p38MAPK and caspase 3 protein expression were significantly increased in pGLV-shRNA-NC group (all P＜0.01).Compared with pGLV-shRNA-NC group,cardiac function was improved,myocardial cell apoptosis index was reduced [(22.35±3.59)％ vs.(31.26±4.45)％,P＜0.05],and the mRNA and protien expressions of p38MAPK and caspase 3 protein expression were decreased in pGLV-shRNA group (all P＜0.05).Conclusions Cardiac dysfunction is associated with p38MAPK-mediated myocardial apoptosis in aldosterone overload MI rats.pGLV-shRNA may inhibit cardiomyocyte apoptosis and improve postMI cardiac function.