目的研究时钟基因-白蛋白D位点结合蛋白(albumin D site-binding protein,DBP)基因在抗Thy-1肾炎模型中随时钟的表达变化,以证明时钟基因参与系膜增殖性肾炎发病进程的调控。方法利用尾静脉注射抗Thy-1抗体制备SD大鼠系膜增殖性肾炎模型;于注射抗体后3 d和5 d的10:00、16:00处死动物,正常对照组注射PBS溶液,并于0 d的10:00、16:00处死。每个时间点需要5只大鼠。利用PAS染色检测系膜增殖和细胞外基质积聚,利用Trizol方法提取RNA,利用q RT-PCR检测DBP基因的表达变化。结果抗Thy-1肾炎模型肾小球自1 d起出现系膜溶解,炎症细胞浸润,至3 d溶解最为明显,5 d溶解消失,系膜细胞开始明显增殖,系膜基质明显增多。抗Thy-1模型的肾小球中,3 d和5 d 10:00的DBP m RNA相对表达量高于正常大鼠,且5 d的16:00时DBP m RNA相对表达量明显高于正常大鼠。但是在肾皮质中,抗Thy-1模型组与正常大鼠相比,DBP m RNA相对表达量并无显著差异。结论 DBP在抗Thy-1肾炎模型肾小球中表达上调,提示时钟基因参与了系膜增殖性肾炎的疾病进展过程。
Objective To investigate expression changes of albumin D site-binding protein(DBP) gene in the anti Thy-1 nephritis model and prove that clock genes are involved in the regulation of the pathogenesis of mesangial proliferative glomerulonephritis(Ms PGN). Methods Ms PGN model was established by tail intravenous injection of anti-Thy-1 antibodies in SD rats, and the animals were scarified at 10 am and 4 pm on day 3 and day 5 after the injection. Animals in normal control group were injected with PBS and scarified at 10 am and 4 pm on day 0. Five rats were used at each time point. PAS staining was used to examine mesangial proliferation and extracellular matrix accumulation. Total RNA was extracted from kidney tissue by Trizol method, then q RT-PCR was used to detect the DBP gene expression. Results On day 1 after the injection, mesangiolysis appeared with inflammatory cell infiltration. On day 5, mesangiolysis disappeared and mesangial cells began to proliferate with extracellular matrix accumulating significantly. DBP m RNA level in glomeruli was higher at 10 am on day 3 and day 5, and it was also higher at 4 pm on day 5 compared to control group. However, there was no significant difference in DBP expression in cortex between Thy-1 model group and control group. Conclusion The up-regulation of DBP in the glomeruli of anti Thy-1 nephritis model implies that the clock genes are involved in the progression of renal disease.