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作 者:王秀梅[1] 张娟[1] 张鸣号[1] 刘爽[1] 李桂忠[1] 曹军[1]
机构地区:[1]宁夏医科大学基础学院病理生理学教研室,宁夏银川750004
出 处:《中国中药杂志》2012年第7期1002-1006,共5页China Journal of Chinese Materia Medica
基 金:宁夏教育厅项目(宁教高[2007]292号);宁夏医科大学重点项目(XZ2009004)
摘 要:目的:观察氧化苦参碱(oxymatrine,OMT)对人脐静脉平滑肌细胞(humans umbilical vein smooth muscle cells,HUSMCs)钙化的影响及其可能机制。方法:以β-甘油磷酸盐诱导HUSMCs钙化,实验分为对照组、钙化组、单纯OMT组、OMT干预高、中、低剂量组。采用Von Kossa染色鉴定细胞钙化,比色法检测细胞钙含量,磷酸苯二钠法测定ALP活性,放射免疫法测定骨钙素(OC)含量,ELISA检测细胞培养液中TGF-β1和细胞内psmad2/3,smad2/3的含量变化,Western blot法检测细胞内Cbfα1蛋白的表达。结果:钙化组与对照组相比平滑肌细胞内可见大量黑色颗粒聚集,钙含量,ALP活性,OC,TGF-β1,smad2/3磷酸化和Cbfα1蛋白的含量均明显增加;OMT干预后钙化指标均下降,TGF-β1,smad2/3磷酸化和Cbfα1蛋白表达量亦明显减少,且高剂量OMT组效应强于中、低剂量组。结论:OMT可有效抑制β-甘油磷酸盐诱导的HUSMCs钙化,且OMT降低TGF-β1,smad2/3磷酸化和Cbfα1蛋白表达可能是OMT抑制HUSMCs钙化的机制之一。Objective: To observe the effect of oxymatrine(OMT) on calcification of humans umbilical vein smooth muscle cells and its underlying mechanism.Method: Human umbilical vein smooth muscle cells(HUSMCs) were calcified by β-giycerophosphosphate(β-GP) and then divided into 6 groups: the control group,the calcification group,the pure OMT group,and lower,middle and higher-dosage OMT groups.Cell calcification were observed by Von Kossa staining,calcium content in HUSMCs were determined by the colorimetric method,the alkaline phosphatase(ALP) activity in HUSMCs were determined by phenyl diphosphate-2-sodium,the osteocalcin(OC) level in HUSMCs were determined by radioimmunossay,the transforming growth factor-β1(TGF-β1) level in HUSMC culture medium and the content changes in psmad2/3 and smad2/3 were determined by the ELISA method,and the expression of Core binding factor α 1(Cbfα1) protein in HUSMCs were determined by western blot method.Result: Compared with the control group,the calcification group showed a great number of black granules among the smooth muscle cells and significant increase in the content of calcium and OC and the activity of ALP;OMT intervention can decrease the content of calcium,OC,TGF-β1,psmad2/3 and Cbfα1 and the activity of ALP.And high-dosage OMT group had better effect than middle and low-dosage groups.Conclusion: OMT can effectively inhibit β-GP-induced HUSMC calcification and its effect on reducing TGF-β1,psmad2/3 and Cbfα1 may be one of its mechanisms in inhibiting HVSMC calcification.
关 键 词:血管钙化 氧化苦参碱 人脐静脉血管平滑肌细胞
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