miR-30b在高磷诱导的大鼠血管平滑肌细胞钙化中的作用  

Role of miR-30b in calcification of rat vascular smooth muscle cells in⁃duced by high phosphorus

作　　者：周薇[1] 徐金升[1] 白亚玲[1] 张慧然[1] 何雷[1] 张胜雷[1] 张东雪[1] ZHOU Wei;XU Jin-sheng;BAI Ya-ling;ZHANG Hui-ran;HE Lei;ZHANG Sheng-lei;ZHANG Dong-xue(Department of Nephrology,The Forth Hospital of Hebei Medical University,Shijiazhuang 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：《中国病理生理杂志》2021年第1期54-59,共6页Chinese Journal of Pathophysiology

基　　金：河北省临床医学优秀人才培养项目(冀财社[2019]139号);河北省医学技术跟踪项目(No.G2018050);河北省重点研发计划项目(No.20377704D)。

摘　　要：目的:观察微小RNA-30b(miR-30b)在高磷诱导的大鼠血管平滑肌细胞(VSMCs)钙化中的作用及其机制。方法:将SD大鼠随机分为假手术(sham)组和慢性肾衰竭所致血管钙化(CRF+VC)组。采用硝酸银染色检测血管钙化情况;免疫组织化学法检测成骨转录因子Runx2的表达。体外培养原代大鼠胸主动脉VSMCs并分为正常组和高磷组。采用邻甲酚酞络合酮比色法及茜素红染色检测VSMCs的钙化情况;ELISA法测定VSMCs中碱性磷酸酶(ALP)活性;RT-qPCR测定VSMCs中miR-30b及Runx2 mRNA的表达;Western blot检测Runx2蛋白的表达。为进一步验证miR-30b对VSMCs的作用,转染miR-30b模拟物(mimic-30b),检测VSMCs钙化、Runx2表达和ALP活性的变化。结果:与sham组比较,CRF+VC组大鼠胸主动脉钙盐沉积和Runx2表达均显著增加(P<0.05);相关分析显示,Runx2与血管钙化呈正相关(r=0.971,P<0.05)。与正常组比较,高磷诱导的大鼠VSMCs钙化增加(P<0.05),Runx2表达及ALP活性均显著升高(P<0.05),而miR-30b水平显著降低(P<0.05)。转染mimic-30b后,miR-30b的表达显著升高(P<0.05),VSMCs钙化程度、Runx2表达及ALP活性均显著降低(P<0.05)。结论:高磷可诱导大鼠VSMCs钙化,可能是通过抑制miR-30b表达、促进细胞表型转化实现的。AIM:To investigate the effect of microRNA-30b(miR-30b)on high phosphorus(HP)-induced calcification of vascular smooth muscle cells(VSMCs)and its mechanism.METHODS:Male SD rats were randomly divided into sham operation(sham)group and chronic renal failure-induced vascular calcification(CRF+VC)group.The vascular calcification was detected by silver nitrate staining,and the expression of osteogenic transcription factor Runx2 was detected by immunohistochemistry.Primary VSMCs from rat thoracic aorta were divided into normal(N)group and HP group.The calcification of the cells was analyzed by Alizarin red staining and alkaline phosphatase(ALP)test.The expression of miR-30b and Runx2 mRNA was detected by RT-qPCR,and the protein expression of Runx2 was detected by Western blot.To further verify the effect of miR-30b on VSMCs,the cell calcification,Runx2 expression and ALP activity were also detected after miR-30b mimic(mimic-30b)transfection.RESULTS:Compared with sham group,the calcium deposition of thoracic aorta and the expression of Runx2 in CRF+VC group were significantly increased(P<0.05).Correlation analysis showed that Runx2 was positively correlated with vascular calcification.Compared with N group,the calcification of VSMCs,the Runx2 expression and the ALP activity in HP group were significantly increased(P<0.05),while the expression of miR-30b was significantly decreased(P<0.05).The expression of miR-30b was increased significantly after the VSMCs was transfected with mimic-30b(P<0.05).Subsequently,cell calcification,Runx2 expression and ALP activity were decreased significantly(P<0.05).CONCLUSION:High phosphorus induces the calcification of rat VSMCs by inhibiting the expression of miR-30b and subsequently promoting the phenotypic transformation of the cells.

关 键 词：微小RNA-30b 高磷 血管钙化 血管平滑肌细胞 表型转化 

分 类 号：R543[医药卫生—心血管疾病] R363[医药卫生—内科学]