CAJ | 학술논문

目的：探讨外源性 H 2 S对柯萨奇病毒B3引起的大鼠乳鼠心肌细胞凋亡的影响及其作用机制。方法培养SD大鼠乳鼠原代心肌细胞,建立病毒性心肌炎(viral myocarditis,VMC)体外模型,随机分为正常组、模型组、模型组+50μmol/L GYY4137组、正常组+50μmol/L GYY4137组。Hoechst 33342染色法及流式细胞术分别检测心肌细胞凋亡,TBA 法及酶标法分别检测各组细胞丙二醛(MDA)、超氧化物歧化酶(SOD)的表达,Western blot 法检测各组细胞Bcl-2、Bax、Cleaved caspase-3蛋白的表达。结果与模型组比较,给予50μmol/L GYY4137后,心肌细胞凋亡明显好转(P<0.01)；50μmol/L GYY4137能使模型组心肌细胞 MDA 显著降低、SOD明显增加(均P<0.05)；给予50μmol/L GYY4137后,模型组心肌细胞Bcl-2蛋白表达增加、Bax蛋白表达明显降低(均P<0.05)、Cleaved caspase-3蛋白表达显著降低(P<0.05)；单纯给予50μmol/L GYY4137对正常心肌细胞上述各项检测指标无明显影响(均P>0.05)。结论GYY4137可显著改善柯萨奇病毒B3引起的大鼠乳鼠心肌细胞凋亡,其作用机制与抑制 Bcl-2/Bax/Cleaved caspase-3信号通路激活有关。
목적：탐토외원성 H 2 S대가살기병독B3인기적대서유서심기세포조망적영향급기작용궤제。방법배양SD대서유서원대심기세포,건립병독성심기염(viral myocarditis,VMC)체외모형,수궤분위정상조、모형조、모형조+50μmol/L GYY4137조、정상조+50μmol/L GYY4137조。Hoechst 33342염색법급류식세포술분별검측심기세포조망,TBA 법급매표법분별검측각조세포병이철(MDA)、초양화물기화매(SOD)적표체,Western blot 법검측각조세포Bcl-2、Bax、Cleaved caspase-3단백적표체。결과여모형조비교,급여50μmol/L GYY4137후,심기세포조망명현호전(P<0.01)；50μmol/L GYY4137능사모형조심기세포 MDA 현저강저、SOD명현증가(균P<0.05)；급여50μmol/L GYY4137후,모형조심기세포Bcl-2단백표체증가、Bax단백표체명현강저(균P<0.05)、Cleaved caspase-3단백표체현저강저(P<0.05)；단순급여50μmol/L GYY4137대정상심기세포상술각항검측지표무명현영향(균P>0.05)。결론GYY4137가현저개선가살기병독B3인기적대서유서심기세포조망,기작용궤제여억제 Bcl-2/Bax/Cleaved caspase-3신호통로격활유관。
Objective To investigate the effect of exogenous H2 S on coxsackie virus B3 (CVB3 )-induced apoptosis of neo-natal rat myocardial cells and its action mechanism.Methods The primary SD neonatal rat myocardial cells were cultured and used for establishment of the in vitro model of viral myocarditis(VMC).They were randomly divided into normal group,model group,model group +50μmol/L GYY4137,normal group+50μmol/L GYY4137.Hoechst 33342 staining and flow cytometry were used to detect myocardial apoptosis,the TBA method and enzyme linked immunosorbent assay to detect the levels of ma-londialdehyde(MDA)and superoxide dismutase(SOD)and Western blot to measure the expression of Bcl-2,Bax and Cleaved caspase-3 proteins in cells in each group.Results Compared with the model group,treatment with 50μmol/L GYY4137 signif-icantly improved the myocardial apoptosis(P<0.01),reduced the level of MDA(P<0.05),and increased the level of SOD in myocardial cells(P<0.05);it conspicuously increased the expression level of Bcl-2 protein,decreased the expression levels of Bax and Cleaved caspase-3 proteins(P<0.05).Moreover,50μmol/L GYY4137 had no effects on these indexes in normal car-diomyocytes(P>0.05).Conclusion GYY4137 can significantly ameliorate the CVB3-induced apoptosis of cardiomyocytes by inhibiting the activation of Bcl-2/Bax/Cleaved caspase-3 signal pathway.