高磷对大鼠胸主动脉血管环中骨形成蛋白2表达的影响

发布时间：2018-05-04 18:00

本文选题：血管平滑肌细胞 + β-甘油磷酸　；参考：《临床心血管病杂志》2017年01期

【摘要】：目的:探讨骨形成蛋白-2(Bone morphogenetic protein 2,BMP-2)在高磷诱导的大鼠胸主动脉血管环钙化中的作用。方法:选取8~10周龄健康雄性SD大鼠,体外培养大鼠胸主动脉血管环,血管环按随机数字表法随机分为2组:正常对照组和高磷组。血管环培养7d和14d后,采用von Kossa染色及邻甲酚酞络合酮比色法检测大鼠胸主动脉血管环钙化情况;免疫组织化学方法检测血管环BMP-2的表达。体外采用组织块贴壁法培养原代大鼠胸主动脉平滑肌细胞,利用10mmol/Lβ-甘油磷酸制备血管平滑肌细胞钙化模型,细胞随机分为2组:正常对照组、高磷组(10mmol/Lβ-甘油磷酸)。采用茜素红染色及邻甲酚酞络合酮比色法检测细胞钙化情况,RT-PCR和Western blot方法检测细胞培养7d和14d的BMP-2mRNA及蛋白表达,并观察短时间内不同时间点BMP-2蛋白表达情况。结果:体外血管环培养7d、14d后,与正常对照组相比,高磷组钙含量明显增加(P0.05);与7d高磷组血管环钙含量相比,14d高磷组血管环钙含量明显增加(P0.05)。免疫组织化学结果显示,与正常对照组相比,高磷组BMP-2表达增加(P0.05);与7d高磷组相比,14d高磷组血管环钙含量明显增加(P0.05)。细胞培养7d、14d后,与正常对照组相比,高磷组钙盐沉积及钙含量明显增高(P0.05);RTPCR和Western Blot显示,与正常对照组相比,高磷组BMP-2mRNA及蛋白表达增加。进一步动态观察BMP-2蛋白的表达变化,结果显示正常对照组及高磷组BMP-2蛋白表达随时间延长呈增强趋势(P0.05)。结论:BMP-2可能参与了高磷诱导的血管钙化的发生发展。
[Abstract]:Aim: to investigate the role of bone morphogenetic protein-2 bone morphogenetic protein 2 (BMP-2) in calcification of thoracic aortic rings in rats induced by high phosphorus. Methods: healthy male SD rats aged 8 ~ 10 weeks were selected and cultured in vitro. The rings were randomly divided into two groups: normal control group and high phosphorus group. After cultured for 7 and 14 days, the calcification of thoracic aortic rings in rats was detected by von Kossa staining and orthophenolphthalein complexone colorimetry, and the expression of BMP-2 in vascular rings was detected by immunohistochemistry. Primary rat aortic smooth muscle cells were cultured by tissue mass adherence in vitro. Vascular smooth muscle cells (VSMCs) calcified by 10mmol/L 尾 -glycerophosphoric acid (10mmol/L 尾 -glycerophosphate) were randomly divided into two groups: normal control group and high phosphorous group (10 mmol / L 尾 -glycerophosphate). The calcification of cells was detected by alizarin red staining and o-cresol phthalein complex ketone colorimetry. RT-PCR and Western blot methods were used to detect the expression of BMP-2mRNA and protein at 7 and 14 days of cell culture, and the expression of BMP-2 protein was observed at different time points in a short period of time. Results: compared with the normal control group, the calcium content in the high phosphorus group was significantly higher than that in the control group for 14 days, and the calcium content in the vascular ring in the high phosphorus group was significantly higher than that in the high phosphorus group on the 7th day after 14 days of culture, and the calcium content in the vascular ring in the high phosphorus group was significantly higher than that in the high phosphorus group on the 14th day. The results of immunohistochemistry showed that compared with the normal control group, the expression of BMP-2 increased in the high phosphorus group and the calcium content in the vascular ring of the high phosphorus group on the 14th day was significantly higher than that in the high phosphorus group on the 7th day. Compared with the normal control group, the calcium salt deposition and calcium content in the high phosphorus group were significantly higher than those in the control group. The expression of BMP-2mRNA and protein in the high phosphorus group was higher than that in the normal control group, and the expression of BMP-2mRNA and protein in the high phosphorus group was significantly higher than that in the control group. The expression of BMP-2 protein was observed dynamically. The results showed that the expression of BMP-2 protein in normal control group and high phosphorus group increased with time. Conclusion: BMP-2 may be involved in the development of vascular calcification induced by high phosphorus.
【作者单位】：河北医科大学第四医院肾内科;
【基金】：河北省自然科学基金资助项目(No:H2012206157) 河北省科技计划项目(No:16397733D);河北省科技计划项目(No:20150351);河北省科技计划项目(No:20150310)
【分类号】：R692;R54

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