Sclerostin／Lrp4调节血管平滑肌细胞钙化的机制研究  

Mechanism of Sclerostin/Lrp4 in the regulation of vascular smooth muscle cell calcification

作　　者：王健[1] 邱小波 盛子桐 徐天华[1] 杜银科[1] 王力宁[1] 姚丽[1] Wang Jian;Qiu Xiaobo;Sheng Zitong;Xu Tianhua;Du Yinke;Wang Lining;Yao Li.(Department of Nephrology, the First Affiliated Hospital of China Medical University, Shenyang 110001, China)

机构地区：[1]中国医科大学附属第一医院肾内科,沈阳110001

出　　处：《中华肾脏病杂志》2018年第3期208-213,共6页Chinese Journal of Nephrology

摘　　要：目的探讨sclerostin／Lrp4在高磷诱导血管平滑肌细胞（vascular smooth musclecell，VSMC）钙化过程中的可能机制及银杏叶提取物的保护作用。方法提取SD大鼠主动脉血管平滑肌细胞并鉴定，分为正常对照组，高磷诱导钙化组（10mmol／L β-甘油磷酸钠＋50μg／ml抗坏血酸），高磷诱导钙化＋银杏叶提取物干预组（10mmol／L β-甘油磷酸钠＋50μg／ml抗坏血酸＋0．5mg／ml银杏叶提取物），培养基诱导干预14d。VONKOSSA染色、茜素红染色检测各组VSMC钙化情况，实时定量PCR检测β-catenin、骨钙素（bone gamma carboxyglutamic acid containing proteins，BGP）的mRNA表达，Western印迹检测sclerostin、Lrp4的蛋白表达。结果VONKOSSA染色、茜素红染色均显示，与正常对照组相比，高磷诱导VSMC钙化组有明显的钙盐沉积，银杏叶提取物干预组钙盐沉积较钙化组明显减少。实时定量PCR结果显示高磷诱导VSMC钙化组B—catenin、BGP的mRNA表达明显高于正常对照组（均P〈0．05）。银杏叶提取物干预组B．catenin、BGP的mRNA表达明显低于钙化组（均P〈0．05）。Western印迹结果显示，高磷诱导VSMC钙化组sclerostin蛋白表达明显高于正常对照组（P〈0．05），Lrp4蛋白表达明显低于正常对照组（P〈0．05）。银杏叶提取物干预组sclerostin蛋白表达明显低于钙化组（P〈0．05），Lrp4蛋白表达明显高于钙化组（P〈0．05）。结论高磷可通过激活Wnt／β-catenin信号通路诱导VSMC钙化，Sclerostin／Lrp4参与了高磷诱导的VSMC钙化过程，而银杏叶提取物可通过抑制Wnt／β．catenin信号通路减轻高磷诱导的VSMC钙化。Objective To investigate the possible mechanism of sclerostin/Lrp4 in calcification of VSMC induced by high phosphorus and the protective effect of Ginkgo biloba extract. Methods Aortic vascular smooth muscle cells （VSMCs） of SD rats were extracted and identified. VSMCs were divided into normal control group, high phosphorus induced calcification group （10 mmol/L β-glyeerophosphate＋50 μg/ml ascorbic acid）, and high phosphorus induced calcification＋Ginkgo biloba extract intervention group （10 mmol/L β-glyeerophosphate＋50 μg/ml ascorbie acid＋0.5 mg/ml GBE）, cultured in different mediums for 14 days. Vonkossa staining and alizarin red staining were used to detect the calcification of VSMCs. The mRNA level of BGP was detected by real time PCR, and the protein expressions of sclerostin and Lrp4 were detected by Western blot. Results Compared with normal control group, vonkossa staining and alizarin red staining showed significant calcium deposition in calcification group, Compared with calcification group, calcium salt deposition was significantly reduced in GBE treatment group. Real time PCR resuhs showed β-cateniu and BGP mRNA expressions in VSMC calcification group were higher than those in normal control group （P〈 0.05）. mRNA expressions of β-catenin and BGP in GBE treatment group were lower than those in calcification group （all P 〈 0.05）. Compared with normal control group, the protein expression of sclerostin was increased, but the protein expression of Lrp4 was decreased in calcified group （all P 〈 0.05）. Compared with calcification group, the protein expression of sclerostin decreased and the protein expression of Lrp4 increased in GBE treatment group （all P 〈 0.05）. Conclusions High phosphorus can induce VSMC calcification by activating Wn/β-catenin signaling pathway. Sclerostin/Lrp4 is involved in hyperphosphineinduced VSMC calcification. GBE can reduce the high phosphorus induced VSMC calcification by regulating the Wnt/β-catenin signaling pathway.

关 键 词：肾功能不全 慢性 钙质沉积 LDL受体相关蛋白质类 骨硬化蛋白 Wnt/β-连环蛋白 银杏叶提取物 

分 类 号：R692[医药卫生—泌尿科学]