酸性环境抑制高磷诱导大鼠VSMCs钙化的机制研究  

Mechanisms of acidic environment inhibiting vascular smooth muscle cell calcification induced by high phosphorus in rats

作　　者：耿同会[1] 徐金升[1] 韩迎迎[1] 白亚玲[1] 张俊霞[1] 崔立文[1] 张胜雷[1] Tong-hui Geng;Jin-sheng Xu;Ying-ying Han;Ya-ling Bai;Jun-xia Zhang;Li-wen Cui;Sheng-lei Zhang(Department of Nephrology,the Forth Hospital of Hebei Medical University,Shijiazhuang,Hebei 050011,China)

机构地区：[1]河北医科大学第四医院肾内科,河北石家庄050011

出　　处：《中国现代医学杂志》2018年第17期1-5,共5页China Journal of Modern Medicine

基　　金：河北省自然科学基金(No:H2012206157);河北省科技计划项目(No:16397733D)

摘　　要：目的探讨酸性环境对高磷诱导的大鼠血管平滑肌细胞(VSMCs)钙化的影响及其机制。方法体外分离培养大鼠VSMCs,采用免疫细胞化学法鉴定。将VSMCs按随机数字表法分为正常对照组、高磷+p H 7.4组、高磷+p H 7.1组。刺激4 d后,采用逆转录聚合酶链反应和Western blot检测活化T细胞核因子c1(NFATc1)、Runt相关转录因子2(Runx2)基因和蛋白的表达。刺激14 d后,对各组细胞进行钙化染色、钙含量和碱性磷酸酶(ALP)活性测定。结果与正常对照组比较,高磷+p H 7.4组的钙含量、ALP活性、Runx2和NFATc1表达升高(P<0.05);与高磷+p H 7.4组比较,高磷+p H 7.1组的钙含量、ALP活性、Runx2和NFATc1表达降低(P<0.05)。相关性分析发现,NFATc1蛋白表达水平与ALP活性、Runx2蛋白表达水平呈正相关(P<0.05)。结论酸性环境可以抑制高磷诱导的大鼠VSMCs钙化,其机制可能是通过降低NFATc1表达,抑制VSMCs表型转化来实现的。Objective To explore the effect and possible mechanisms of acidification on high phosphorusinduced vascular smooth muscle cell（VSMC） calcification in rats. Methods VSMCs were isolated from rat aorta, and randomly divided into control group and high phosphorus group. The high phosphorus group was further settled into two subgroups, i.e. high phosphorus＋p H 7.1 group and high phosphorus＋p H 7.4 group, which were treated with β-glycerophosphate, and acidified by HCl to adjust the p H. The m RNA expressions of nuclear factor of activated T-cells cytoplasmic 1（NFATc1） and Runt-related transcription factor 2（RUNX2） in VSMCs were detected by RT-PCR after stimulation for 4 days. The activity of alkaline phosphatase（ALP） and calcium deposition were tested after culture for 14 days. Results Compared to the control group, calcium depositon and ALP activity were significantly increased in the high phosphorus＋p H 7.4 group（P〈0.05）, and the expressions of NFATc1 and RUNX2 were upregulated as well（P〈0.05）. Meanwhile, the calcification level and the expressions of RUNX2 and NFATc1 in the high phosphorus＋p H 7.1 group were lower than those in the high phosphorus＋p H 7.4 group（P〈0.05）. NFATc1 expression level was positively correlated with ALP activity and RUNX2 expression level（P〈0.05）. Conclusions Acidic environment can inhibit VSMC calcification induced by high phosphorus in rats, its mechanism is possibly by downregulation of NFATc1 expression and prevention of VSMC phenotypic transformation.

关 键 词：血管钙化 血管平滑肌细胞 活化T细胞转录因子c1 Runt相关转录因子2 慢性肾衰竭 

分 类 号：R692.5[医药卫生—泌尿科学]