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  

Extracelluar pH values influence high-phosphorus-induced VSMCs calcification mediated by BMP-2 signaling pathway

作　　者：张慧然[1] 徐金升[1] 郭利萍[1] 白亚玲[1] 张胜雷[1] 张俊霞[1] 崔立文[1] 

机构地区：[1]河北医科大学第四医院肾内科,050011

出　　处：《天津医药》2016年第7期861-864,共4页Tianjin Medical Journal

基　　金：河北省自然科学基金资助项目(H2012206157)

摘　　要：目的探讨不同pH 值环境下骨形态发生蛋白2（BMP-2）信号通路对高磷诱导大鼠血管平滑肌细胞（VSMCs）钙化的影响.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,取第4 代VSMCs 进行实验,设正常对照组、pH7.4＋高磷组、pH7.1＋高磷组和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（AKP）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、茜素红染色钙化结节增多,BMP-2、Smad1、Runx2 mRNA 表达及AKP 活性均增高（均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、茜素红染色钙化结节减少,BMP-2、Smad1、Runx2 mRNA 表达及AKP 活性均降低（均P 〈 0.05）,pH7.7＋高磷组大鼠VSMCs 的钙含量增加、茜素红染色钙化结节增多,BMP-2、Smad1、Runx2 mRNA 表达及AKP 活性均增高（均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VSMCs 表型转化.Objective To explore the effect of different pH values on calcification of rat vascular smooth muscle cells (VSMCs) through bone morphogenetic protein (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 in vitro, and then identified by immunocytochemistry. The VSMCs were randomly divided into 4 groups by random sampling method:normal group (pH 7.4), pH7.4+high phosphorus group, pH 7.1+high phosphorus group, and pH 7.7+high phosphorus group. Calcium deposition and alkaline phosphatase (AKP) activity were measured by alizarin red staining and enzyme linked immunosorbent assay. The expressions of BMP-2, Smad1 and Runx2 mRNA were detected by RT-PCR. Results Compared with the control group, the calcification staining was increased in pH 7.4+high phosphorus group, calcium content was increased and expressions of BMP-2, Smad1, Runx2 mRNA and AKP activity were also increased (P<0.05). While compared with the pH 7.4+high phosphorus group, calcification staining, calcium content, expressions of BMP-2, Smad1, Runx2 mRNA and AKP activity were decreased in pH 7.1+high phosphorus group (P<0.05). The calcification staining, calcium content, expressions of BMP-2, Smad1, Runx2 mRNA and AKP activity were increased in pH 7.7+high phosphorus group (P<0.05). Conclusion The extracellular acidic environment (pH 7.1) can inhibit high-phosphotus-induced VSMCs calcification, whereas extracellular alkaline environment (pH 7.7) induce high-phosphotus-induced VSMCs calcification. The mechanism is presumably that VSMCs calcification is induced by influencing BMP-2 pathway, which may be mediated by VSMCs phenotype transdifferentiation of BMP-2 signaling pathway.

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 

分 类 号：R692.5[医药卫生—泌尿科学]