CAJ | 학술논문

目的：观察龙胆苦甙（Gentiopicroside，GPS）后处理对离体心肌细胞缺血/再灌注损伤（ischemia and reperfusion injury，I/R）的拮抗作用及其可能的机制。方法采用SD大鼠乳鼠培养心肌原代细胞，用缺氧复氧模型模拟缺血再灌注（stimulated ischemia reperfusion，SI/R）。实验分为正常对照组（Control＋ScrambleRNA＋Veh）；缺氧复氧组（I/R＋ScrambleRNA＋Veh）；缺氧复氧＋龙胆苦甙后处理组（I/R＋ScrambleRNA＋GPS 组）以及缺氧复氧＋AktSiRNA＋龙胆苦甙后处理组（I/R＋AktSiRNA＋GPS）。采用化学法缺氧复氧模型，缺氧2 h，复氧4 h。复氧前给予GPS药物，检测心肌细胞乳酸脱氢酶（LDH）以及TUNEL染色确定心肌细胞的损伤程度以及抑制Akt表达后GPS的保护作用变化。结果与I/R＋ScrambleRNA＋Veh组相比，I/R＋ScrambleRNA＋GPS组LDH明显降低（P＜0.01），TUNEL染色阳性率增加减少（P＜0.01），Akt/Gsk3β信号通路磷酸化程度明显增加（P＜0.01）。与I/R＋ScrambleRNA＋GPS组相比，SI/R＋SiAktRNA＋GPS组，LDH活性显著增加（P＜0.01），TUNEI染色阳性率增加（P＜0.01），Gsk3β磷酸化程度减弱（P＜0.01）。结论 GPS后处理对I/R大鼠心肌具有保护作用，其作用机制与AKT/Gsk3β信号通路的活化有关。
목적：관찰룡담고대（Gentiopicroside，GPS）후처리대리체심기세포결혈/재관주손상（ischemia and reperfusion injury，I/R）적길항작용급기가능적궤제。방법채용SD대서유서배양심기원대세포，용결양복양모형모의결혈재관주（stimulated ischemia reperfusion，SI/R）。실험분위정상대조조（Control＋ScrambleRNA＋Veh）；결양복양조（I/R＋ScrambleRNA＋Veh）；결양복양＋룡담고대후처리조（I/R＋ScrambleRNA＋GPS 조）이급결양복양＋AktSiRNA＋룡담고대후처리조（I/R＋AktSiRNA＋GPS）。채용화학법결양복양모형，결양2 h，복양4 h。복양전급여GPS약물，검측심기세포유산탈경매（LDH）이급TUNEL염색학정심기세포적손상정도이급억제Akt표체후GPS적보호작용변화。결과여I/R＋ScrambleRNA＋Veh조상비，I/R＋ScrambleRNA＋GPS조LDH명현강저（P＜0.01），TUNEL염색양성솔증가감소（P＜0.01），Akt/Gsk3β신호통로린산화정도명현증가（P＜0.01）。여I/R＋ScrambleRNA＋GPS조상비，SI/R＋SiAktRNA＋GPS조，LDH활성현저증가（P＜0.01），TUNEI염색양성솔증가（P＜0.01），Gsk3β린산화정도감약（P＜0.01）。결론 GPS후처리대I/R대서심기구유보호작용，기작용궤제여AKT/Gsk3β신호통로적활화유관。
Objective To investigate the protective effect of post-treatment with gentiopicroside (GPS) on myocardial ischemia reperfusion and to elucidate the underlying mechanism. Methods We cultivated the isolated neonatal rat cadiomyocyte to suffer the hypoxia/reoxygenation. The cells suffered control, stimulated ischemia/reperfusion(SI/R), SI/R+ScrambleRNA+GPS and SI/R+SiAktRNA+GPS respectively. After 2 h hypoxia following 4 h reoxygenation (GPS was given before reoxygenation), the LDH as well as the TUNEL positive staining was measured. We used the AktSiRNA to silence the Akt expression, and observed the changes of LDH and TUNEL positive staining. Results Compared with the Control group, the LDH amount and TUNEL positive level was enhanced in the SI/R group. The GPS post-treated measurement managed to decline the LDH amount as well as the TUNEL positive ratio and to upregulate the phosphorylation rate of Akt/Gsk3β signaling pathway compared with the SI/R. The protective effect of GPS was blurred after depleted the Akt expression by using SiRNA. Conclusion Post-treatment with gentiopicroside can significantly reduce myocardial ischemia/reperfusion injury partially via the Akt/Gsk3βsignaling pathway.