【摘要】 目的以乳鼠心肌细胞缺氧/复氧模型模拟在体心肌缺血再灌注损伤,观察附子多糖后处理对缺氧/复氧后心肌细胞的作用及机制。方法建立乳鼠心肌细胞缺氧/复氧模型,将乳鼠心肌细胞分为正常对照组、缺氧/复氧组、缺氧后适应组和附子多糖组。缺氧/复氧组给予心肌细胞缺氧3 h后复氧6 h;缺氧后适应组在细胞缺氧3 h后,复氧前即给予3个循环的5 min复氧/5 min缺氧,随后复氧6 h;附子多糖组在缺氧3 h后,将心肌细胞换入含附子多糖浓度为10 mg.mL-1的培养液中,常规培养6 h。MTT法检测细胞活力,流式细胞仪测定细胞内钙离子浓度和心肌细胞凋亡率,检测细胞培养液中乳酸脱氢酶(LDH)和肌酸激酶(CK)的活性。结果与缺氧/复氧组比较,经附子多糖后处理,可以增加心肌细胞存活率(P<0.01),减少LDH(P<0.01)和CK(P<0.05)的释放,降低细胞内钙离子浓度(P<0.05),有效抑制心肌细胞的凋亡(P<0.05)。结论附子多糖对缺氧/复氧后心肌细胞产生保护效应,机制与其抑制钙超载、减轻线粒体的损伤有关。更多还原

【Abstract】 Objective To investigate the effect and mechanism of monkshood polysaccharide,which was isolated from Radix Aconiti Lateralis Preparata,against hypoxia-reoxygenation injury in neonatal rat cardiomyocytes with hypoxia-reoxygenation(H/R),which was used for mimic in vivo ischemia-reperfusion injury(I/R).Methods Rat myocardial cell model of hypoxia-reoxygenation injury was established.Neonatal rat cardiomyocytes were divided into four groups:control,H/R,postconditioning,and monkshood polysaccharide postconditioning groups.The cells in H/R group were incubated primarily in hypoxic buffer solution for 3 hours firstly and then were incubated for 6 hours in normal culture medium.The cells in postconditioning group were given tree cycles of 5-min reoxygenation and 5-min hypoxia prior to 6 hours of reoxygenation.The cells in monkshood polysaccharide postconditioning group were cultured in medium added with 10mg.mL-1 monkshood polysaccharide for 6 hours after 3 hours of hypoxia.Cell count was measured with MTT method.The calcium concentration and cell apoptosis of cardiomyocytes were measured by flow cytometry.LDH and CK activities in cell culture fluid were also measured.Results As compared with H/R group,monkshood polysaccharide postconditioning increased cell survival rate apoptotic rate(P < 0.01),reduced the release of LDH(P < 0.01) and CK(P < 0.05),decreased the intracellular calcium concentration(P < 0.05),and inhibited the apoptosis of myocardial cells(P < 0.05).Conclusion These data suggest that monkshood polymaccharide postconditioning can protect myocardial cells from H/R injury.The mechanism is probably related to maintaining calcium concentration and lessening the mitochondria injury.更多还原