摘要
目的 探讨泛素羧基末端水解酶-1（ubiquitin carboxy-terminal hydrolase-1,UCH-L1）在高糖诱导的体外足细胞及糖尿病肾病大鼠模型体内足细胞中的表达。方法 以25mmol/L高糖刺激体外足细胞0、12、24h后,提取组织蛋白,采用Western bolt法检测UCH-L1的表达。20只8~10周龄Wistar大鼠,随机分为对照组和糖尿病肾病模型（STZ）组并造模,各10只,定期检测体重、血糖变化,处死大鼠取肾脏组织,HE染色和PAS染色观察肾脏形态学改变,采用免疫组化PV9000两步法染色以WT1定位足细胞,检测足细胞内UCH-L1的表达。结果 在不同时间高糖处理后,足细胞UCH-L1蛋白表达量随时间延长明显升高（P〈0.01）;体重、血糖检测及HE、PAS染色证明糖尿病肾病模型构建成功,STZ组足细胞（WT1定位）内UCH-L1呈阳性,而对照组几乎无明显表达。结论 分子生物学和形态学两种方法联合证明高糖可诱导体内外足细胞中UCH-L1的表达升高。
Purpose To explore the expression of UCH-L1 in podocytes stimulated with high glucose and streptozotocin （STZ） -in- duced diabetic nephropathy rat model. Methods Podocytes were stimulated with high glucose （25 mmol/L D-glucose） for 0, 12, 24 hours, and then proteins were extracted and analyzed for the expression of UCH-L1 using Western blot. Wistar rats were randomly di- vided into two groups： control group （Control, n = 10）, STZ-indueed diabetic nephropathy rat model group （STZ, n = 10） and made the injection respectively. The changes of weights and blood glucose were tested at special times. Kidney tissues from killed rats were stained with hematoxylin-eosin （HE） and periodic acid sehiff （PAS） to observe morphological changes. Immunohistochemistry of PV 9000 two-step analysis was performed to detect the expression of UCH-L1 and to mark podocytes with WT1. Results After stimulated with high glucose, the expression of UCH-L1 significantly increased with time （P 〈 0. 01 ）. Both the changes of weights and blood glu- cose and the results of the HE and PAS stains confirmed the successful construction of STZ-induced diabetic nephropathy rat model. In the STZ group, UCH-L1 was positively expressed in podocytes （ marked with WT1 ） , however, it was nearly negative in control group. Conclusion High glucose can induce the upregulation of UCH-L1 in vitro and in vivo, which is proved by molecular biological method and morphological method.
出处
《临床与实验病理学杂志》
CAS
CSCD
北大核心
2016年第9期975-979,共5页
Chinese Journal of Clinical and Experimental Pathology
基金
国家自然科学基金(81274049)
山东省医药卫生科技发展计划(2014WS0182)
山东省高等学校科技计划(J15LK11)
山东省潍坊市科学技术发展计划(2014WS047)
山东省自然科学基金(ZR2015HL091)
