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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 the calcification of human umbilical vein smooth muscle cells (HUSMCs) and its possible mechanism. Methods: The calcification of HUSMCs was induced by β-glycerophosphate. The experiment was divided into control group, calcification group, OMT group and OMT intervention group. Cell calcification was determined by Von Kossa staining, calcium content was assayed by colorimetry, ALP activity was measured by phenyl-disodium phosphate method, and osteocalcin (OC) content was measured by radioimmunoassay. The levels of TGF-β1 and psmad2 / 3, smad2 / 3 content changes, Western blot detection of intracellular Cbfα1 protein expression. Results: Compared with the control group, a large number of black granules were found in the calcification group and the calcium content, ALP activity, OC, TGF-β1, smad2 / 3 phosphorylation and Cbfα1 protein content were significantly increased in the calcified group compared with the control group Decreased TGF-β1, smad2 / 3 phosphorylation and Cbfα1 protein expression was significantly reduced, and high-dose OMT group stronger than the middle and low dose group. CONCLUSION: OMT can effectively inhibit β-glycerophosphate-induced calcification of HUSMCs. OMT may be one of the mechanisms by which OMT can inhibit the calcification of HUSMCs by decreasing TGF-β1, Smad2 / 3 phosphorylation and Cbfα1 protein expression.