论文部分内容阅读
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 detect the expression of fractalkine (Fkn) CX3C in puromycin aminonucleoside (PAN) nephropathy and further elucidate the pathogenesis of nephropathy and provide a new target for the treatment of nephropathy point. Methods: PAN was administered from the jugular vein of Wistar rats, and the control group was given the same amount of normal saline. The changes of urinary protein and renal infiltration of inflammatory cells induced by PAN were observed at different time points. The animals were sacrificed on day 10 to prepare kidney homogenates and frozen sections for RT PCR and immunohistochemistry respectively to observe the expression of Fk mRNA and protein in renal tissues at different time points after injections of PAN. 1|Â (ILl|Â) stimulation of cultured tubular epithelial cells observed Fkn expression. Results: Urinary protein began to increase on the 5th day after PAN injection and continued to increase until day 10, accompanied by an increase of CD4 + T cells, CD8 + T cells and monocytes / macrophages in the renal interstitium. Both RT PCR and immunohistochemistry showed that Fkn was increased on day 7 and day 10, mainly on glomeruli on day 3, and mainly expressed in renal tubular epithelial cells later. The tubular epithelial cells cultured in vitro 1 h after IL 1 β stimulation began to express Fk mRNA and peaked at 4 ~ 6 h. Conclusions: Fkn was first observed in PAN nephropathy model, which was characterized by first appearance in the glomerulus, then migration to the renal tubules, and earlier than leukocyte infiltration and proteinuria in the kidney tissue. Tip Fkn may be another important related molecules in the pathogenesis and progression of nephropathy.