肾动脉钙化促进2型糖尿病肾病大鼠进行性肾功能损伤

基础医学与临床 ›› 2017, Vol. 37 ›› Issue (2): 217-223.

肾动脉钙化促进2型糖尿病肾病大鼠进行性肾功能损伤

覃春美¹,龚财判²,魏晓³,吴蔚桦¹,刘建⁴,欧三桃¹

1. 泸州医学院附属医院肾病内科
2. 泸州医学院附属医院
3. 泸州医学院
4. 泸州医学院附属医院肾内科

收稿日期:2016-03-07 修回日期:2016-07-01 出版日期:2017-02-05 发布日期:2017-01-16
通讯作者: 欧三桃 E-mail:422216756@qq.com
基金资助:
泸州市人民政府-四川医科大学科技战略合作项目

Renal artery calcification promotes the progressive renal damage of type 2 diabetic nephropathy rats

Received:2016-03-07 Revised:2016-07-01 Online:2017-02-05 Published:2017-01-16

摘要/Abstract

摘要： 目的探讨肾动脉钙化对2型糖尿病肾病大鼠肾功能的影响。方法将大鼠随机分为对照组(CON组)、糖尿病肾病组(DN组)及DN合并肾动脉血管钙化组(DN+VC组)。DN组和DN+VC组大鼠在高脂高糖饮食基础上腹腔注射链脲菌素（STZ）制备2型糖尿病肾病（diabetic nephropathy，DN）大鼠模型, 造模成功后DN+VC组肌注维生素D3及尼古丁灌胃，分别于实验第8、12和16周末处死大鼠，用Von Kossa染色观察大鼠肾动脉钙盐沉积，Ca2+试剂盒检测肾动脉钙含量，免疫荧光双染观察肾动脉α-SMA/BMP2表达，荧光定量PCR检测肾动脉BMP2 mRNA水平等判断肾动脉钙化程度。检测大鼠血清尿素氮、肌酐、胱抑素C及24 h尿蛋白水平，HE染色观察肾组织病理改变。结果 DN组及DN+VC组肾动脉的钙盐沉积及钙含量、BMP2蛋白及基因表达均较同时间点CON组明显增加， DN+VC组较DN组增加更明显（P<0.05）。DN组和DN+VC组大鼠血清尿素氮、肌酐、胱抑素C及24 h尿蛋白随时间进展逐渐升高，且明显高于同时间点CON组（P<0.05）。与DN组相比，DN+VC组大鼠仅胱抑素C水平明显升高（P<0.05），24 h尿蛋白量在第16周时明显增加（P<0.05）。DN组大鼠肾组织病理损伤呈进行性加重，DN+VC组大鼠的病理改变较DN组明显加重。肾动脉钙含量与血清尿素氮、肌酐、胱抑素C、24 h尿蛋白以及BMP2 mRNA呈正相关(P< 0.01)。结论肾动脉钙化的发生及严重程度可能参与和促进糖尿病肾病的进展。

关键词: 肾动脉, 血管钙化, 糖尿病肾病, 骨形态蛋白2

Abstract: Objective To explore the effects of renal artery calcification on the renal function in type 2 diabetic nephropathy rats. Methods Rats were randomly divided into control group(CON group), diabetic nephropathy group (DN group)and DN with vascular calcification group (DN+VC group). Rats of group DN and DN+VC were were fed with high sugar and fat diet and injected with streptozotocin（STZ）into abdominal cavity to induce a model of type 2 diabetes. After diabetic models were successfully made, rats of group DN+VC were treated by vitamin D3 plus nicotine. The rats were sacrificed at 8, 12 and16 week respectively and the pathologic change to the renal artery were detected by von Kossa staining. The calcium content were detected by calcium assay kit and double immunofluorescence staining and real-time polymerase chain reaction (RT-qPCR) were applied to detect the protein and gene expression levels of BMP2 in the renal artery. Measure the levels of blood urea nitrogen (BUN),serum creatinine (Scr),cystatin C(Cys C) and 24 hour urinary protein (24-h UA)respectively at the 8th,12th and 16th weeks. Histopathology of kidney was assessed by hematoxylin/eosin staining . Results The deposition of black granules, the calcium content and the protein and gene expression levels of BMP2 in DN group were significantly higher than those in group CON and lower than DN+VC group at each time points（P<0.05）. The BUN, Scr, Cys C and 24-h UA in group DN and group DN+VC were gradually increased in 8th,12th and 16th weeks, and were higher than those in group CON（P<0.05）. Compared with the DN group, only the level of Cys C at each time point and the level of 24-h UA in 16th week in DN+VC group were significantly higher（P<0.05）. The pathological damages of the kidney in group DN showed a continual worsening trend and the pathological changes of the kidney in group DN+VC were more serious than group DN. Calcium content was positively correlated with the increased serum BUN, Scr, Cys C, 24- h UA and BMP2 mRNA (all P< 0.01). Conclusion The occurrence and severity of renal artery calcification may participate in and promote the progression of DN.

Key words: Vascular calcification, Diabetic nephropathy, Renal artery, Bone Morphogenetic Protein 2

覃春美龚财判魏晓吴蔚桦刘建欧三桃. 肾动脉钙化促进2型糖尿病肾病大鼠进行性肾功能损伤[J]. 基础医学与临床, 2017, 37(2): 217-223.

[1]	邱维张薇陶建瓴王海云郑法雷李雪梅. 倍半萜内酯减轻糖尿病小鼠胰岛素抵抗及早期肾损伤[J]. 基础医学与临床, 2018, 38(9): 1292-1297.
[2]	李晓明王青竹石婧鞠瑞朱蕾李娟郭磊叶菜英. 小檗碱改善动脉粥样硬化小鼠的血管炎性反应和钙化[J]. 基础医学与临床, 2018, 38(2): 163-168.
[3]	李爱玲彭睿孙艳彭惠民易红张政. 长链非编码RNA-1700020I14Rik对高糖培养下肾系膜细胞纤维化的影响[J]. 基础医学与临床, 2017, 37(6): 781-785.
[4]	文利陈蕾孙一瑄段青蕊魏礼江周琴彭惠民张政. 柚皮素通过抑制TGF-β1/smad信号传导通路缓解糖尿病肾病大鼠肾损伤[J]. 基础医学与临床, 2016, 36(7): 896-901.
[5]	陈艳霞曹莉萍房向东涂卫平. 幽门螺杆菌感染与肾脏相关性疾病[J]. 基础医学与临床, 2014, 34(8): 1150-1153.
[6]	周敏张晓良. 微小RNAs在糖尿病肾病中的作用[J]. 基础医学与临床, 2014, 34(5): 711-714.
[7]	黄炜徐玲徐勇周雪琴彭娟. SUMO化修饰对高糖诱导的大鼠肾系膜细胞IκB/NF-κB信号的影响[J]. 基础医学与临床, 2014, 34(12): 1689-1693.
[8]	刘瑞霞郭兵肖瑛石明隽王圆圆李晓颖张国忠. SnoN和Smad2/3信号蛋白与糖尿病大鼠肾小管-间质纤维化的关系[J]. 基础医学与临床, 2010, 30(9): 944-951.
[9]	刘淑霞张玉军张景坤刘青娟唐丽娟段惠军. 核转录因子κΒ/环氧化酶-2参与糖尿病肾病大鼠肾细胞增殖[J]. 基础医学与临床, 2008, 28(9): 944-948.
[10]	方开云娄晶磊肖瑛石明隽桂华珍郭兵张国忠. Snail1在糖尿病大鼠肾组织中表达增加[J]. 基础医学与临床, 2008, 28(9): 957-963.
[11]	史永红段惠军王丽晖. 氯沙坦对糖尿病大鼠肾皮质信号转导和转录活化因子1、3表达的影响[J]. 基础医学与临床, 2006, 26(1): 102-102.