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2020 年第 8 期 第 15 卷

雷公藤甲素对大鼠血管钙化过程中骨形态发生蛋白2和Runt相关转录因子2蛋白表达的影响

Effect of Triptolide on expression of BMP-2 and Runt2 protein in rat vascular calcification

作者:裴昱强丁耀东王睿刘晓丽申华葛海龙 

英文作者:Pei Yuqiang Ding Yaodong Wang Rui Liu Xiaoli Shen Hua Ge Hailong

单位:首都医科大学附属北京安贞医院心内科100029

英文单位:Department of Cardiology Beijing Anzhen Hospital Capital Medical University Beijing 100029 China 

关键词:血管钙化;雷公藤甲素;骨形态发生蛋白2;Runt相关转录因子2

英文关键词:Vascularcalcification;Triptolide;Bonemorphogeneticprotein-2;Runt-relatedtranscriptionfactor2

  • 摘要:
  • 目的 探究雷公藤甲素对大鼠血管钙化过程中骨形态发生蛋白2(BMP-2)Runt相关转录因子2(Runx2)蛋白表达的影响。方法 选取30只雄性清洁级SD大鼠随机分成对照组、钙化模型组和雷公藤干预组,每组10只。对照组给予肌肉注射0.9%氯化钠注射液和单纯花生油灌胃;钙化模型组给予肌肉注射维生素D3和尼古丁溶于花生油灌胃处理,制作胸主动脉钙化模型;雷公藤干预组在钙化模型组处理的基础上加用雷公藤甲素干预。喂养4周后处死大鼠,取胸主动脉,用von Kossa染色及苏木精-伊红染色反映血管钙化情况,检测血管钙含量和碱性磷酸酶(ALP)活性,蛋白质印迹法检测BMP-2Runx2蛋白表达。结果 von Kossa染色结果 显示,与钙化模型组相比,雷公藤干预组血管中膜棕黑色颗粒显著减少,显示血管钙化程度较轻。苏木精-伊红染色结果 显示,雷公藤干预组大鼠动脉组织血管内膜破坏不明显,中膜钙化灶钙化程度较钙化模型组明显减轻。雷公藤干预组血管组织中钙含量和ALP活性明显低于钙化模型组[(31.9±2.3)μmol/g protein(47.6±3.6)μmol/g protein(82.4±0.9)U/g protein(149.6±2.7)U/g protein](均P0.05)。蛋白质印迹法检测结果 显示,与钙化模型组相比,雷公藤干预组血管组织中BMP-2Runx2的表达水平显著下调。结论 雷公藤甲素具有减轻血管钙化的作用,这种作用可能与雷公藤甲素抑制参与血管平滑肌细胞成骨转分化的重要信号通路上BMP-2/Runx2蛋白的表达有关。

  • Objective To evaluate the effect of Triptolide on expression of bone morphogenetic protein-2 (BMP-2) and Runt2 protein in rat vascular calcification model. Methods Totally 30 male SD rats were randomly divided into 3 groups, with 10 rats in each group (control group, the calcification group and the intervention group). Control group was given volume of 0.9% sodium chloride Vitamin D3 and nicotine dissolved in peanut oil to establish thoracic aorta vascular calcification model. Vascular calcification was detected by von Kossa staining; calcium content and the activity of alkaline phosphatase (ALP) were detected. Western blot was used to detect the protein expression of BMP-2 and Runt2. Results Von Kossa staining showed that compared with the calcification model group, there was a significant decrease of brownish black granules in the middle membrane of blood vessels in the intervention group of Tripterygium wilfordii, indicating that the degree of calcification was lighter. The Results  of HE staining showed that there was no obvious destruction of vascular intima in the intervention group of Tripterygium wilfordii; the calcification degree of middle membrane calcification was significantly lower than that in the calcification model group. Compared with the calcification model group, the calcium content and ALP activity in the vascular tissue of Tripterygium wilfordii intervention group decreased significantly(31.9±2.3)μmol/g protein vs (47.6±3.6)μmol/g protein, (82.4±0.9)U/g protein vs (149.6±2.7)U/g protein](all P0.05. The Results  of Western blotting showed that the expression of BMP-2 and Runx2 in vascular tissue of Tripterygium intervention group was significantly lower than that of calcification model group. Conclusion Triptolide can reduce on vascular calcification, which may be related to triptolide inhibiting the expression of BMP-2/Runt2 protein in the important signaling pathway involved in osteogenesis and transdifferentiation of vascular smooth muscle cells.

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