不规则趋化蛋白在嘌呤霉素氨基核苷肾病模型中的表达  

Expression of fractalkine in rats with puromycin aminonucleoside nephropathy

作　　者：朱春芳[1] 王磊 白冰[1] 欧周罗[1] 

机构地区：[1]复旦大学上海医学院生物化学与分子生物学系,上海200032 [2]复旦大学附属中山医院肾内科

出　　处：《肾脏病与透析肾移植杂志》2003年第3期234-239,共6页Chinese Journal of Nephrology,Dialysis & Transplantation

摘　　要：目的 :检测CX3C族趋化因子不规则趋化蛋白 (fractalkine,Fkn)在嘌呤霉素氨基核苷 (puromycinaminonucleoside,PAN)肾病中的表达 ,进一步阐明肾病的发病机制 ,为肾病的治疗提供新的靶点。　 方法 :从Wistar大鼠的颈静脉给予PAN ,对照组给予等量生理盐水 ,在不同的时间点观察PAN引起的尿蛋白改变和肾脏炎性细胞浸润情况 ,并于第 1、3、5、7、1 0天处死动物 ,制作肾组织匀浆和冰冻切片 ,分别用于RT PCR和免疫组化研究 ,观察在PAN注射的不同时间点肾组织中FknmRNA和蛋白质的表达情况 ;同时在体外用白介素 1 β(IL 1 β)刺激培养的肾小管上皮细胞观察Fkn的表达。　 结果 :PAN注射后第 5天尿蛋白开始升高 ,持续增加直至第 1 0天 ,同时伴有肾间质中CD4+ T细胞 ,CD8+ T细胞和单核 /巨噬细胞的增加。RT PCR和免疫组化均显示Fkn在第 7、第 1 0天表达升高 ,第 3天主要分布在肾小球 ,以后主要在肾小管上皮细胞表达。用IL 1 β刺激后 1h体外培养的肾小管上皮细胞即开始表达FknmRNA ,在 4～ 6h达到高峰。　 结论 :本文首次观察到Fkn在PAN肾病模型中表达增高 ,其特征为先出现于肾小球 ,后移行至肾小管 ,并早于肾组织中白细胞浸润及蛋白尿的发生。提示Fkn可能是肾病发病和进展过程中的另一重要的相关分子。Objective:To investigate the expression and potential effect of CX3C chemokine fractalkine (Fkn) in rats with puromycin aminonucleoside (PAN) nephropathy and further illustrate the mechanism of renal disease. Methodology:Male Wistar rats (150～170 g body weight) were injected with PAN (9 mg/100g BW) via left jugular vein, control rats received an equal volume of saline. Those rats were sacrificed 1, 3, 5, 7 and 10 days after the injection ( n =6 rats in each group). The expression of Fkn mRNA and protein were detected by reverse transcription polymerase chain reaction (RT PCR) and immunohistochemistry, respectively. The leukocytes infiltration in rats with PAN nephropathy was examined by immunohistochemistry. At the same time, the expression of Fkn in NRK 52E cells stimulated with IL 1β(10 ng/ml) in vitro were observed. Results:At the 5 th day after a single injection of PAN, urinary protein of 24hrs increased until the tenth day. At the same time, there was significant increase in renal interstitial infiltrates including CD4 +, CD8 + cells and monocytes/macrophages. Both the expression of Fkn mRNA and protein were increased at 7 th and 10 th day after injection with PAN. The protein of Fkn at third day stained transiently in glomeruli and thereafter distributes mostly in proximal tubular epithelial cells. The expression of Fkn mRNA began to increase at one hour in NRK 52E cells stimulated with IL 1β, peaked at 4～6hrs and decreased thereafter. Conclusion:These results indicate that the mRNA and protein of Fkn enhance obviously in rats with PAN nephropathy. The expression of Fkn is moved from glomeruli to tubulointerstitium, which preceded the appearance of leukocytes infiltration and urinary protein. Fkn may be another strong candidate for directing leukocytes infiltration in PAN nephropathy.

关 键 词：不规则趋化蛋白 嘌呤霉素氨基核苷 肾病 动物模型 生理盐水 大鼠 发病机制 

分 类 号：R692.3[医药卫生—泌尿科学]