摘要
目的探讨山药多糖(CYPS)对心肌梗死大鼠心肌细胞凋亡的抑制作用及其机制。方法大鼠心肌细胞随机分为空白对照(NC)组、缺氧/复氧(H/R)组、H/R+75 mg/L CYPS组、H/R+150 mg/L CYPS组、H/R+300 mg/L CYPS组,噻唑蓝(MTT)法检测心肌细胞存活率;流式细胞术检测心肌细胞凋亡率;Western印迹法检测心肌细胞细胞周期蛋白依赖性激酶(CDK)4、活化半胱氨酸天冬氨酸蛋白酶(Cleaved caspase)-3、成纤维细胞生长因子(FGF)9蛋白表达。比较FGF9过表达及沉默FGF9表达对H/R损伤心肌细胞存活率及凋亡率的影响。观察CYPS不同剂量组及FGF9表达对心肌细胞中谷胱甘肽过氧化物酶(GSH-Px)、超氧化物歧化酶(SOD)含量的影响。结果与NC组相比,H/R组心肌细胞存活率显著降低(P<0.05),CDK4蛋白水平及GSH-Px、SOD含量显著降低(P<0.05),而细胞凋亡率与Cleaved caspase-3蛋白水平显著升高(P<0.05);与H/R组相比,CYPS不同剂量组心肌细胞存活率显著升高(P<0.05),CDK4蛋白水平及GSH-Px、SOD含量显著升高(P<0.05),而细胞凋亡率与Cleaved caspase-3蛋白水平显著降低(P<0.05);CYPS可促进H/R损伤心肌细胞中FGF9表达水平升高(P<0.05);FGF9过表达对H/R损伤心肌细胞具有保护作用;沉默FGF9逆转CYPS对H/R损伤心肌细胞的保护作用。结论CYPS可通过上调FGF9表达对H/R损伤的心肌细胞发挥保护作用,其作用机制可能与抑制心肌细胞凋亡及增强抗氧化能力有关。
Objective To investigate the inhibitory effect of Chinese polysaccharide from Yam(CYPS)on cardiomyocyte apoptosis in rats with myocardial infarction and its mechanism.Methods Rat cardiomyocytes were randomly divided into NC,H/R,H/R+75 mg/L CYPS,H/R+150 mg/L CYPS,H/R+300 mg/L CYPS groups.MTT assay was used to detect survival rate of cardiomyocytes.Apoptosis rate of cardiomyocytes was detected by flow cytometry.Protein expression of CDK4,Cleaved caspase-3 and fibroblast growth factor(FGF)9 were detected by Western blot.The effects of FGF9 over-expression and silencing FGF9 expression on survival rate and apoptosis rate of cardiomyocyte injury by H/R were compared.The effects of different doses of CYPS and FGF9 on the contents of GSH-PX and SOD in cardiomyocytes were observed.Results Compared with NC group,the survival rate of cardiomyocytes in H/R group was significantly lower(P<0.05),the levels of CDK4 and GSH-PX and SOD were significantly lower(P<0.05),but the apoptotic rate and the level of Cleaved caspase-3 were significantly increased(P<0.05).Compared with H/R group,the survival rate of cardiomyocytes in different dose of CYPS groups was significantly increased(P<0.05),and the expression level of CDK4 and GSH-PX and SOD were significantly increased(P<0.05),the apoptotic rate and the expression level of Cleaved caspase-3 were significantly lower(P<0.05).CYPS could promote the expression of FGF9 in H/R injuring cardiomyocytes(P<0.05).Over-expression of FGF9 had a protective effect on H/R injuring cardiomyocytes.Silencing FGF9 reversed the protective effect of CYPS on H/R injuring cardiomyocytes.Conclusions The Chinese polysaccharide from Yam could protect the cardiomyocytes damaged by H/R by up-regulating the expression of FGF9,and its mechanism might be related to inhibiting cardiomyocyte apoptosis and enhancing antioxidant capacity.
作者
王静
洪炳哲
张习敬
葛鲁敏
任海勤
WANG Jing;HONG Bing-Zhe;ZHANG Xi-Jing(Department of Cardiology,General Hospital,Guizhou Panjiang Investment Holding Group Co.,Ltd,Liupanshui 553530,Guizhou,China)
出处
《中国老年学杂志》
CAS
北大核心
2020年第8期1707-1711,共5页
Chinese Journal of Gerontology
基金
国家自然科学基金(31070998)。
