目的:研究霍山石斛多糖抗心肌缺血-再灌注损伤的作用。方法:采用Langendorff离体灌流系统建造大鼠离体缺血-再灌注损伤模型。实验分为正常对照组(不停灌处理)、模型组(停灌20 min)及霍山石斛多糖低、中、高浓度(停灌前灌流已配制浓度为0.1、0.5、1.0 mg/L的KH液10 min)组。利用生物多导记录仪连续测定血流动力学参数。通过TTC染色检测心肌梗死面积。并用ELISA法检测冠脉流出液中肌酸激酶(CK)和乳酸脱氢酶(LDH)的活性。ELISA法检测心肌组织中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量、还原型谷胱甘肽/氧化型谷胱甘肽(GSH/GSSG)比率、白介素6(IL-6)和肿瘤坏死因子α(TNF-α)的含量。利用HE染色和TUNEL法分别检测心肌细胞的坏死和凋亡。Western blot检测心肌组织中GSK-3β的磷酸化水平。结果:与正常对照组比较,模型组出现严重的心肌坏死和极高的凋亡率,心功能下降,冠脉流出液中LDH和CK水平显著升高,心肌组织中MDA、IL-6和TNF-α含量显著升高,SOD的活性与GSH/GSSG比值均显著降低(P<0.01)。与模型组比较,霍山石斛多糖中、高剂量组的心肌坏死程度、梗死面积和细胞凋亡率显著降低,心功能显著改善;冠脉流出液中LDH、CK水平逐渐降低;心肌组织中MDA、TNF-α及IL-6含量显著降低,SOD活性和GSH/GSSG比值显著升高(P<0.01)。高剂量霍山石斛多糖可显著降低心肌细胞凋亡指数,提高心肌组织中GSK-3β磷酸化水平。结论:霍山石斛多糖具有显著的抗心肌缺血-再灌注损伤作用,其作用机制可能与激活GSK-3β信号通路发挥抗氧化、抗炎和抗凋亡有关。 Objective: To study the effect of Dendrobium huoshanense polysaccharides on myocardial ischemia-reperfusion injury. Methods: Langendorff isolated perfused system was used to establish the rat model of ischemia / reperfusion injury in vitro. The experiment was divided into normal control group (non-stop irrigation), model group (stop-irrigation for 20 min) and Dendrobium huoshanense at low, medium and high concentration (perfusion before perfusion has been prepared at concentrations of 0.1,0.5,1.0 mg / L of KH Liquid 10 min) group. Continuous measurement of hemodynamic parameters using a bio-guide recorder. Myocardial infarction area was measured by TTC staining. The activities of creatine kinase (CK) and lactate dehydrogenase (LDH) in coronary effluent were detected by ELISA. The activities of superoxide dismutase (SOD), malondialdehyde (MDA), reducing glutathione / glutathione (GSH / GSSG), interleukin 6 (IL- And tumor necrosis factor alpha (TNF-alpha) content. Cardiomyocyte necrosis and apoptosis were detected by HE staining and TUNEL method respectively. Western blot was used to detect the phosphorylation of GSK-3β in myocardium. Results: Compared with the normal control group, severe myocardial necrosis and high apoptosis rate were found in the model group. The cardiac function was decreased. The levels of LDH and CK in coronary effluent were significantly increased. The levels of MDA, IL-6 and TNF -α, and the activity of SOD and the ratio of GSH / GSSG were significantly decreased (P <0.01). Compared with the model group, the myocardial necrosis, infarction area and apoptosis rate of the Dendrobium huoshanense polysaccharide group were significantly decreased, the cardiac function was significantly improved; the levels of LDH and CK in the coronary effluent fluid decreased gradually; the levels of MDA, The levels of TNF-α and IL-6 were significantly decreased and the activities of SOD and GSH / GSSG were significantly increased (P <0.01). High doses of D. huoshanense polysaccharide can significantly reduce the apoptosis index of cardiac myocytes and increase the phosphorylation of GSK-3β in myocardium. CONCLUSION: Dendrobium huoshanense polysaccharides have significant anti-myocardial ischemia-reperfusion injury and its mechanism may be related to the activation of GSK-3β signaling pathway to exert anti-oxidant, anti-inflammatory and anti-apoptotic effects.