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出 处:《中国实验方剂学杂志》2013年第22期226-231,共6页Chinese Journal of Experimental Traditional Medical Formulae
基 金:湖南省高校创新平台开放基金项目(09K107);湖南省教育厅十二.五重点建设学科基金项目;湖南省心脑血管天然药物研究重点实验室基金项目
摘 要:目的:研究马齿苋总黄酮(PTF)对缺氧/复氧致心肌细胞损伤的影响及机制。方法:利用体外培养的H9c2心肌细胞建立缺氧/复氧模型。将培养的心肌细胞分为5组:正常对照组、缺氧/复氧损伤模型组、缺氧/复氧损伤加10 mg·L-1PTF组、缺氧/复氧损伤加20 mg·L-1PTF组、缺氧/复氧损伤加40 mg·L-1PTF组。MTT法检测心肌细胞存活率,荧光分光光度法测定心肌细胞内钙离子浓度,比色法测定心肌细胞培养液中一氧化氮(NO)浓度,流式细胞术检测心肌细胞凋亡率、RT-PCR检测心肌细胞凋亡基因Caspase-3 mRNA的表达。结果:与A/R组比较,PTF组(终质量浓度为10,20,40 mg·L-1)均可明显提高缺氧/复氧损伤的心肌细胞存活率,降低NO浓度、细胞内钙离子浓度、细胞凋亡率[(18.25±2.64,P<0.05),(13.83±1.52,11.21±1.76,P<0.01)],PTF组可有效减少缺氧/复氧损伤心肌细胞内Caspase-3 mRNA的表达。结论:PTF可通过抑制心肌细胞内钙超载、减轻过量NO的细胞毒性作用、下调Caspase-3表达,从而保护心肌细胞。Objective:To study effect of portalaca total flavone (PTF) on H9c2 myocardial cell inj induced by anoxia/reoxygenation (A/R) and its possible mechanism.Method:Myocytes cell model of A/R injury was established in vitro cultured.The cell viability was measured with MTT,Intra cellular calcium concentration was determined by Fluorospectrop hotometry,nitric oxide (NO) concentration were detected using colori metricmet hod,using flow cytometry to detect myocardial cell apoptosis rate,myocardial cell apoptosis gene expression of Caspase-3 mRNA detected by RT-PCR.Result:Compared with the A/R group,after pretreatment with 10,20 and 40 mg ·L-1 PTF,PTF groups can significantly improve anoxia/reoxygenation injury of myocardial cell survival rate,reduce the concentration of NO,intracellular calcium ion concentration,the cell apoptosis rate (P 〈 0.05,P〈0.01),PTF groups can effectively reduce hypoxia/reoxygenation injury of myocardial Caspase-3 mRNA expression in the cell.Conclusion:PTF can protect the myocardial cells with in hibit myocardial intracellular calcium overload,reducing excessive NO cytotoxic effect and downgrading of caspase 3 mRNA expression.
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