血清组织非特异性碱性磷酸酶与终末期肾脏病大鼠冠脉钙化的关联研究  

Study on the association between serum tissue non-specific alkaline phosphatase and coronary artery calcification in rats with end-stage renal disease

作　　者：王玉静 李慧 张文强 弥娜 侯云飞 谢晓莉 WANG Yu-jing;LI Hui;ZHANG Wen-qiang(Department of Cardiology,the First Affiliated Hospital of Air Force Military Medical University,Xi'an Shaanxi 710016,China;Department of Cardiology,the 986th Hospital of the Air Force,Xi'an Shaanxi 710016,China.)

机构地区：[1]空军军医大学第一附属医院心内科,陕西西安710016 [2]空军第九八六医院心内科,陕西西安710016

出　　处：《临床和实验医学杂志》2021年第4期337-341,共5页Journal of Clinical and Experimental Medicine

基　　金：陕西省社会发展领域项目(编号:2020SF-263)。

摘　　要：目的探究血清组织非特异性碱性磷酸酶(TNAP)与终末期肾脏病大鼠冠脉钙化的关联及机制。方法选择42只大鼠按随机数字表法分为对照组(健康大鼠等量0.9%氯化钠溶液灌胃,n=14)、ESRD模型组(终末期肾脏病大鼠模型等量0.9%氯化钠溶液灌胃,n=14)和治疗组[终末期肾脏病大鼠模型7.2 mg·kg^(-1)·d^(-1)呋塞米片悬浊液灌胃,n=14],于实验第5~7周进行灌胃。连续治疗3周后,分别采用酶联免疫吸附检测血清TNAP水平含量;通过彩色多普勒超声心动图检测冠脉钙化情况;蛋白印迹分析检测骨钙蛋白、骨桥蛋白和胶原蛋白Ⅱ的表达;实时逆转录酶-聚合酶链式反应(RT-qPCR)分析检测Runt相关转录因子2(Runx2)、成纤维细胞生长因子-23(FGF-23)、骨形态发生蛋白2(BMP-2)mRNA表达情况。线性回归分析进行TNAP与冠脉钙化评分相关性分析。结果ESRD模型组大鼠TNAP水平、左心室舒张末期内径、左心室收缩末期内径、主动脉瓣面积、二尖瓣面积、骨钙蛋白、骨桥蛋白、胶原蛋白Ⅱ的蛋白表达以及Runx2、FGF-23、BMP-2 mRNA表达均显著高于对照组,差异均有统计学意义(P<0.05),治疗组大鼠TNAP水平、左心室舒张末期内径、左心室收缩末期内径、主动脉瓣面积、二尖瓣面积、骨钙蛋白、骨桥蛋白、胶原蛋白Ⅱ的蛋白表达以及Runx2、FGF-23、BMP-2 mRNA表达均显著低于ESRD模型组,差异均有统计学意义(P<0.05);血清TNAP与冠脉钙化评分呈显著正相关(r=0.566,P<0.05)。结论终末期肾脏病治疗后,大鼠冠脉钙化程度的减轻,伴随着大鼠血管内皮中TNAP水平以及骨钙蛋白、骨桥蛋白、胶原蛋白Ⅱ表达量的降低,且血清TNAP与冠脉钙化具有显著正相关关系。Objective To investigate the relationship and mechanism between non-specific alkaline phosphatase of vascular endothelial tissue and coronary artery calcification in rats with end-stage renal disease.Methods Forty-two rats were selected and divided into the control group(healthy rats are given intragastrically with the same amount of normal saline,n=14),ESRD model group(end-stage renal disease rat model with equal amount of normal saline gavage,n=14)and treatment group(end-stage renal disease rat model with 7.2 mg·kg^(-1)·d^(-1) furosemide tablet suspension by intragastric administration,n=14)according to random number table method,and gavage was performed on the 5th to 7th week of the experiment,after 3 weeks of continuous treatment,Enzyme-linked immunosorbent assay is used to detect serum TNAP levels;to detect coronary artery calcification by color Doppler echocardiography;Western blot analysis was used to detect the expression of osteocalcin,osteopontin and collagen Ⅱ;Real-time reverse transcriptase-polymerase chain reaction(RT-qPCR)analysis was used to detect the expression of Runt-related transcription factor 2(Runx2),fibroblast growth factor-23(FGF-23),bone morphogenetic protein 2(BMP-2)mRNA.The correlation between TNAP and coronary artery calcification score was analyzed by linear regression.Results Compared with the control group,ESRD model group TNAP level,LVDd,LVDs,AVA,MVA,osteocalcin,osteopontin,collagen Ⅱ protein expression and Runx2,FGF-23,BMP-2 mRNA expression were all significant increased(P<0.05),and the above indicators in the treatment group were significantly lower than those in the ESRD model group(P<0.05);The serum TNAP is significantly positively correlated with coronary artery calcification score(r=0.566,P<0.05).Conclusion After treatment of end-stage renal disease,the reduction in the degree of coronary artery calcification in rats is accompanied by the decrease in the level of tissue non-specific alkaline phosphatase and the expression of osteocalcin,osteopontin,and collagen II in t

关 键 词：大鼠 终末期肾脏病 组织非特异性碱性磷酸酶 骨钙蛋白 骨桥蛋白 冠脉钙化 

分 类 号：R692.5[医药卫生—泌尿科学] R-332[医药卫生—外科学]