摘要
目的以细胞凋亡的线粒体信号通路为着眼点,对附子多糖对缺氧/复氧后心肌细胞的保护机制进行探讨。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 g.L?1的培养液中常规培养6 h。流式细胞仪测定心肌细胞凋亡率和线粒体膜电位,进行凋亡诱导因子(AIF)的Western blotting分析,检测心肌细胞内SOD活性和MDA含量。结果与缺氧/复氧组相比较,附子多糖后处理可以保护心肌细胞SOD活性,减少MDA的生成,阻止线粒体膜电位的下降,抑制AIF自线粒体向胞浆的释放,减少心肌细胞凋亡率。结论附子多糖后处理对缺氧/复氧后心肌细胞的保护机制与其抗氧化损伤,抑制细胞凋亡的线粒体信号途径有关。
OBJECTIVE To use cultured neonatal rat cardiomyocytes with hypoxia-reoxygenation（H/R） to mimic in vivo ischemia-reperfusion injury（I/R）,and to investigate the mitochondrial protective mechanism of Fuzi polymccharide postconditioning in cardiomyocytes against hypoxia-reoxygenation injury.METHODS Rat myocardial cells were divided into four groups： control,H/R,postconditioning and Fuzi polymccharide postcondition groups.The cells in H/R group were incubated primarily in hypoxic buffer solution for 3 hours,thereafter,these cells 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 hypoxic prior to 6 hours of reoxygenation.The cells in Fuzi polymccharide postconditioning group were cultured in medium added with 10 g·L 1 Fuzi polymccharide for 6 hours after 3 hours of hypoxia.Cell apoptosis of cardiomyocytes and mitochondrial membrane potential of cardiomyocytes were measured by flow cytometry.AIF was detected by Western blotting.The SOD activity and MDA content within cardiomyocytes were measured.RESULTS Compared with H/R group,Fuzi polymccharide postconditioning protected the SOD activity,reduced production of MDA,kept mitochondrial membrane potential of cardiomyocytes stable,depressed the translocation of AIF from mitochondria into cytosol,and decreased the cell apoptosis.CONCLUSION These data suggest Fuzi polymccharide postconditioning can protect cardiomyocytes from H/R injury.The mechanism is related to its antioxidation and protection on mitochondria.
出处
《中国现代应用药学》
CAS
CSCD
2012年第4期281-284,共4页
Chinese Journal of Modern Applied Pharmacy
基金
广东省科技计划项目(2008B060600055)
作者简介
作者简介：刘颖，女，博士，副教授Tel：13538789626.E-mail：13322819916@163．com
