天津医药

天津医药 ›› 2016, Vol. 44 ›› Issue (7): 861-864.doi: 10.11958/20150117

• 实验研究 • 上一篇    下一篇

胞外pH值通过BMP-2信号通路影响高磷诱导的血管平滑肌细胞钙化

张慧然， 徐金升， 郭利萍， 白亚玲， 张胜雷， 张俊霞， 崔立文   

  1. 北医科大学第四医院肾内科（邮编 050011）
  • 收稿日期:2015-08-28 修回日期:2016-03-25 出版日期:2016-07-15 发布日期:2016-07-15
  • 通讯作者: 徐金升 E-mail:mailglp@163.com
  • 作者简介:张慧然（1978）， 女， 硕士， 主治医师， 主要从事慢性肾脏病血管钙化方面研究
  • 基金资助:
    河北省自然科学基金资助项目（H2012206157）

Extracelluar pH influence high-phosphorus-induced VSMCs calcification mediated by BMP-2 Signaling Pathway

ZHANG Huiran, XU Jinsheng△, GUO Liping, BAI Yaling, ZHANG Shenglei, ZHANG Junxia, CUI Liwen   

  1. epartment of Nephrology, the Forth Hospital of Hebei Medical University, Shijiazhuang 050011, China
  • Received:2015-08-28 Revised:2016-03-25 Published:2016-07-15 Online:2016-07-15
  • Contact: XU Jinsheng E-mail:mailglp@163.com

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摘要： 目的 探讨骨形态发生蛋白2（bone morphogenetic protein 2，BMP-2）信号通路在不同pH值环境影响高磷诱导的大鼠血管平滑肌细胞（vascular smooth muscle cells, VSMCs）钙化中的作用。方法 选取5～8周龄健康雄性SD大鼠，体外分离培养大鼠胸主动脉VSMCs，采用免疫细胞化学方法鉴定。将VSMCs用随机抽样法分为正常对照组(pH7.4)、pH7.4+高磷组、pH7.1+高磷组及pH7.7+高磷组。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，酶联免疫吸附法检测细胞碱性磷酸酶(ALP)活性，用反转录聚合酶链反应（RT-PCR）法检测BMP-2、Smad1及 Runt 相关转录因子2（Runx2） mRNA的表达。结果 与正常对照组相比，pH7.4+高磷组大鼠VSMCs的钙含量增加（P<0.05）、茜素红染色钙化结节增多，Runx2 mRNA表达及ALP活性均增高（P<0.05）；与pH7.4+高磷组相比，pH7.1+高磷组大鼠VSMCs的钙含量减低（P<0.05）、茜素红染色钙化结节减少，Runx2 mRNA表达及ALP活性均降低（P<0.05），BMP-2、Smad1mRNA表达均降低（P<0.05），pH7.7+高磷组大鼠VSMCs的钙含量增加（P<0.05）、茜素红染色钙化结节增多，Runx2 mRNA表达及ALP活性均增高（P<0.05），BMP-2、Smad1mRNA表达均增高（P<0.05）。结论 胞外酸性环境（pH7.1）抑制高磷诱导的大鼠VSMCs钙化，胞外碱性环境（pH7.7）促进高磷诱导的大鼠VSMCs钙化，其机制可能是通过BMP-2信号通路介导了VSMCs的表型转化。

关键词: 肌细胞, 平滑肌, 氢离子浓度, 骨形态发生蛋白质类, 血管钙化, pH, 骨形态发生蛋白 2 信号通路

Abstract: Abstract：Objective To explore the?effect of different?pH on calcification of rat vascular smooth muscle cells (VSMCs) through the bone morphogenetic protein2 (BMP-2) signaling pathway. Methods Healthy male SD rats aged?5-8-weeks were selected in the study.?VSMCs?from rat thoracic?aorta?were cultured?invitro,?and then identified?by?immunocytochemistry.?The?VSMCs?were randomly?divided into?4 groups by random sampling method: normal group(pH7.4), pH7.4+high phosphorus group, pH7.1+high phosphorus group, and pH7.7+high phosphorus group.Calcium deposition and ALP activity were measured by Alizarin red staining, quantification of calcium and enzyme linked immunosorbent assay.The of BMP-2, Smad1, Runx2 mRNA was detected by RT-PCR .Results Compared with the control group, the pH7.4+ high phosphorus group calcification staining, calcium content, alkaline phosphatase (ALP) activity and the of Runx2 mRNA average increased (P<0.05).While compared with the pH7.4+high phosphorus group, the pH7.1+high phosphorus group calcification staining, calcium content, alkaline phosphatase (ALP) activity and the of BMP-2, Smad1, Runx2 mRNA average decreased (P<0.05), and the pH7.7+high phosphorus group calcification staining, calcium content, alkaline phosphatase (ALP) activity and the of BMP-2, Smad1, Runx2 mRNA average increased (P<0.05).Conclusion Extracelluar acidic environment(pH7.1) can inhibit high-phosphotus-induced VSMCs calcification,whereas extracelluar alkaline environment(pH7.7) induce high-phosphotus-induced VSMCs calcification.The mechanism is presumably that VSMCs calcification is induced by influencing BMP-2 pathway, which may mediate the transdifferentiation of vascular smooth muscle cells.

Key words: Key words：Vascular smooth muscle cells, Vascular calcification, pH, BMP-2 signaling pathway