活性维生素D3 对阿霉素肾病大鼠CD2AP 的影响

Effect of active vitamin D3 on CD2AP in rats with adriamycin-induced nephropathy

摘要:

目的 探讨活性维生素D 类似物骨化三醇对阿霉素（ADR）肾病大鼠足细胞CD2 相关蛋白 （CD2AP）表达的影响。方法 随机法选取60 只Sprasue-Dawley 大鼠，单次尾静脉注射ADR 7.5 mg/kg 复制 ADR 肾病模型。2 周后检测24 h 尿蛋白、血清总蛋白（TP）、血浆白蛋白（ALB）、总胆固醇（TC）、血清 肌酐（Scr）水平。将模型大鼠随机均分为模型组（ADR）、治疗组（ADR+VitD3），另取30 只大鼠作为对照 组。于模型复制成功后2、4、6、8 和10 周检测24 h 尿蛋白。取大鼠肾组织，用Masson 染色观察肾小球的 病理损伤，透射电子显微镜观察足细胞超微结构，免疫组织化学染色和Western blotting 检测CD2AP 定位和 表达。结果 2 周后3 组24 h 尿蛋白、TP、ALB、Scr、TC 比较，差异有统计学意义（P <0.05），提示模型复 制成功。大鼠24 h 尿蛋白随治疗周数延长而减少（P <0.05）。组织病理学结果显示，模型组肾组织大量纤维 增生，肾小管扩张和肾间质纤维化。透射电子显微镜结果显示，模型组肾足细胞结构损伤，足突广泛融合，核 畸形，脂滴聚集；治疗组较模型组有所改善。免疫组织化学染色结果显示，模型组CD2AP 表达较低，活性维 生素D3 治疗后阳性表达面积增加（P <0.05）。各组大鼠CD2AP 表达水平比较，差异有统计学意义（P <0.05）； 模型组降低，治疗组随时间延长表达增强（P <0.05）。结论 活性维生素D3 通过上调CD2AP 表达，减轻足 细胞的损伤，抑制肾组织病理纤维化增生，发挥肾保护功能，减少蛋白尿的产生。

Abstract:

Objective To investigate the effect of active vitamin D analogue (calcitriol) on the expression of CD2AP in podocytes of rats with adriamyicin (ADR) nephropathy. Methods Sixty Sprasue-Dawley rats were selected randomly and injected by single tail vein injection of doxorubicin (7.5mg/kg). The levels of 24h urine protein and total protein (TP), albumin (ALB), total cholesterol (TC) and serum creatinine (Scr) were measured to judge the model success after 2 weeks. These rats were randomly divided into model group (ADR group) and calcitriol treatment group (ADR+VitD3 group). The other 30 rats were used as a blank control group. 2, 4, 6, 8, 10 weeks after the injection of ADR, 24 h urine proteins were detected and kidney tissues were taken. The pathological damages of glomerulus were observed by Masson staining, and the ultrastructure of podocytes were observed by transmission electron microscope. Immunohistochemical staining and western blotting were used to detect the localization and expression of CD2-related proteins. Results There were significant differences in urine protein quantitation, TP, ALB,Scr and TC among the control group, the model group, and the treatment group at 2 weeks (P < 0.05), suggesting that the model was successful. The 24 h urine protein quantitation of rats decreased with the number of weeks of treatment (P < 0.05). Histopathology showed a large amount of fibrous hyperplasia, tubular dilatation and renal interstitial fibrosis in the renal tissue of the model group. Transmission electron microscopy showed that structural damage of the renal foot cells in the model group, extensive fusion of the foot processes, nuclear malformation, and lipid droplet aggregation. The treatment group was better than the model group. The results of immunohistochemistry showed that the expression of CD2AP was lower in the model group than control group, and the positive expression areas were significantly increased after treatment with 1, 25- (OH)2D3 (P < 0.05). Western blotting showed that the expression of CD2AP in each group was significantly different (P < 0.05). and decreased in the model group. The expression of the treatment group was increased with the number of weeks (P < 0.05). Conclusions Active vitamin D3 can upregulate the expression of CD2-related proteins, reduce the damage of podocytes, inhibit the pathological fibrosis and proliferation of renal tissues, and exert renal protective function to reduce the production of proteinuria.