Kindlin-2介导足细胞损伤在局灶节段肾小球硬化症模型小鼠发病中的作用研究  

Kindlin-2 in Mediates Podocyte Injury in Focal Segmental Glomerulosclerosis Mice

作　　者：任凌燕 朱鸣[1] 霍丽霞 王霄一[1] REN Lingyan;ZHU Ming;HUO Lixia;WANG Xiaoyi(Department of Nephrology,Huzhou First People＇s Hospital,HuZhou（313000）,China;Central Laboratory,Huzhou First People＇s Hospital,HuZhou（313000）,China)

机构地区：[1]浙江省湖州市第一人民医院肾内科,湖州313000 [2]浙江省湖州市第一人民医院中心实验室,湖州313000

出　　处：《浙江中西医结合杂志》2018年第9期730-734,816,共7页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：浙江省医药卫生科技计划项目(No.2017KYB639);浙江省湖州市科技计划项目(No.2016GY27)

摘　　要：目的探讨Kindlin-2介导足细胞损伤在局灶节段肾小球硬化症模型小鼠发病中的作用。方法 8周龄雄性BALB/C小鼠分为正常对照组和模型组,各18只。模型组小鼠注射阿霉素(ADR,浓度2mg/m L,剂量10mg/kg鼠重)造模,正常对照组小鼠给予等量的生理盐水,于第4、8、12周每组各处死6只小鼠,检测24h尿蛋白定量、血清白蛋白(ALB)、总胆固醇(TC),光镜观察肾脏组织病理学改变,免疫组织化学技术检测足细胞数量,RT-q PCR技术检测肾脏组织Kindlin-2、肾病蛋白(Nephrin)及Podocin m RNA表达水平。结果与正常对照组比较,模型组4、8、12周24h尿蛋白定量、血总胆固醇(TC)均增加[24h尿蛋白定量:(5.66±1.06)mg/24h比(1.72±0.62)mg/24h、(4.63±1.16)mg/24h比(1.77±0.63)mg/24h、(4.78±0.87)mg/24h比(1.69±0.49)mg/24h;血TC:(8.84±1.30)mmol/L比(2.01±0.49)mmol/L、(7.45±1.27)mmol/L比(2.06±0.49)mmol/L、(6.79±1.38)mmol/L比(2.10±0.54)mmol/L,P均<0.05];血ALB水平降低[(11.98±1.33)g/L比(22.27±1.41)g/L、(12.78±1.47)g/L比(22.65±1.49)g/L、(12.99±1.38)g/L比(23.31±1.49)g/L,P均<0.05];模型组第4周可见部分肾小球出现局灶、节段硬化,伴有足细胞肿胀;第8周部分肾小球出现球性硬化,伴有明显的足细胞增生和肥大;第12周弥漫性肾小球出现球形硬化。模型组足细胞数量随着时间的进展逐渐减少[(77.40±7.56)个/肾小球比(104.00±10.05)个/肾小球,(63.27±8.51)个/肾小球比(103.80±12.23)个/肾小球,(53.15±8.23)个/肾小球比(98.86±8.89)个/肾小球,P均<0.05]。模型组第4周开始Kindlin-2 m RNA水平明显上调[(1.14±0.06)、(1.13±0.08)、(1.45±0.10)比(0.82±0.07),P均<0.05],Podocin m RNA水平明显下调[(1.79±0.08)、(1.64±0.06)、(1.45±0.07)比(2.05±0.05),P均<0.05],第8周开始Nephrin m RNA水平明显下调[(1.42±0.13)、(1.17±0.11)比(2.06±0.20),P均<0.05]。Kindlin-2 m RNA水平与足细胞数量(r=-0.02,P<0.01)、Nephrin m RNA水平(r=-0.8,P<0.01)、Podocin m RNA水平(r=-0.9,P<0.01)�Objective To investigate the mechanism of Kindlin-2 in mediating podocyte injury in focal segmental glomerulosclerosis mice. Methods 8-week-old male BALB/C mice were divided into two groups： the normal control group and the disease model group, each had 18 mice. The model mice were treated with adriamycin（ADR. 2 mg/m L, 10 mg/kg mouse） by tail intravenous injection, and the control mice were given the same amount of saline. Six mice were killed at 4 th, 8 th and 12 th week, respectively. The levels of 24 h urinary protein, serum albumin（ALB）, and total cholesterol（TC） were detected. Pathology of kidney was observed using light microscopy.Number of podocytes was counted by immunohistochemistry method. m RNA expression levels of Kindlin-2, Nephrin and Podocin were tested by real-time quantitative PCR. Results After injected ADR for 4, 8 and 12 weeks, the levels of both 24 h urinary protein and serum TC increased, serum ALB decreaed significantly（P〈0.05）. 24 h urinary protein was（5.66 ±1.06） vs（1.72 ±0.62）,（4.63 ±1.16） vs（1.77 ±0.63）, and（4.78 ±0.87） vs（1.69 ±0.49）mg/24 h;serum TC was（8.84 ±1.30） vs（2.01 ±0.49）,（7.45 ±1.27） vs（2.06 ±0.49）, and（6.79 ±1.38） vs（2.10 ±0.54）mmol/L;serum ALB was（11.98±1.33） vs（22.27±1.41）,（12.78±1.47） vs（22.65±1.49）, and（12.99±1.38） vs（23.31±1.49）g/L. In the 4 th-week model mice, partly focal and segmental glomerulosclerosis with podocyte swelling were observed. In the 8 th-week model mice, spherical glomerulosclerosis with significant proliferation and hypertrophy of podocytes were seen. In the 8 th-week model mice, diffuse spherical glomerulosclerosis were examined. Number of podocytes in the disease mice reduced significantly（P 0.05）,（77.40 ±7.56） vs（104.00 ±10.05）,（63.27 ±8.51） vs（103.80±12.23）, and（53.15±8.23） vs（104.00±10.05）/glomerulus. The m RNA level of Kindlin-2 up-regulated, and Podocin down-regulated significantly from 4 th week（P 0.

关 键 词：小鼠 Kindlin-2 足细胞损伤 局灶节段肾小球硬化症 

分 类 号：R692.6[医药卫生—泌尿科学]