目的 观察紧密连接蛋白在高尿酸血症致大鼠肾损害模型中的表达变化以及非布司他的干预疗效。方法 将SD大鼠分为正常组,高尿酸血症组(模型组),非布司他组(干预组);氧嗪酸联合尿酸诱导制作高尿酸血症大鼠模型,给予非布司他进行干预,分别于6周后检测各组大鼠血中尿素氮(BUN)、血肌酐(Scr)、尿酸(UA)水平,免疫组化及RT-PCR方法检测紧密连接蛋白包括膜周蛋白-1(ZO-1)、跨膜蛋白(occludin) 的表达变化,采用Masson染色检测大鼠肾间质病理改变。结果 6周时,模型组、干预组ZO-1、occludin表达较正常组降低(均P<0.05);干预组ZO-1、occludin表达较模型组增加,差异有统计学意义(均P<0.05),与正常组相比,模型组、干预组RIF指数均增高(均P<0.05),干预组RIF指数低于模型组,高于正常组(均P<0.05)。结论 紧密连接蛋白表达的降低在高尿酸血症肾间质纤维化发展过程中起着举足轻重的作用,并与血尿酸水平及肾功能损害密切相关。非布司他通过降低血尿酸水平,能改善紧密连接蛋白的表达,延缓肾功能损害,起到肾保护作用。

Objective To observe the expression of tight junction protein in hyperuricemia induced renal damage model in rats and the intervention effect of febuxostat. Methods SD rats were randomly divided into three groups: normal control group, model control group, febuxostat treatment group. Hyperuricemia was induced in rats with oxonic acid per time for three times per day, by gavage and combined with uric acid added in drinking water, while febuxostat were administered by gavage in febuxostat treatment group.The blood of rats were collected to analyse the differences of control, model and treatment group on changes of blood urea nitrogen (BUN), creatinine (Cr), uric acid (UA). Immunohistochemistry was used to assay ZO-1 and occludin protein expression and quantitive real time PCR to detect the expression of ZO-1 and occludin in renal tissue of renal interstitial fibrosis model rats induced by hyperuricemia. Paraffin section of kidney was maked and then performed Masson staining to make sure the model is successful. Results At 6 weeks, the expressions of ZO-1 and occludin in the model group and treatment group were lower than those in the normal group (all P<0.05). The expressions of ZO-1 and occludin in the treatment group were higher than those in the model group (all P<0.05). Compared with the normal group, the RIF index in the model group and treatment group were higher (all P<0.05), and the RIF index in the treatment group was lower than that in the model group and higher than that in the normal group (all P<0.05). Conclusion The downregulated expression of ZO-1 and occludin plays a crucial role during the development of hyperuricemia in renal interstitial fibrosis, and are closely related to UA level and renal function impairment. Febuxostat may improve the expression of tight junction by downregurating UA, reduce renal fuction impairment and play a role in renal protection.

目的 探讨2型糖尿病肾病(DN)患者肾组织中STOML2的表达及作用。方法 免疫组化检测临床2型糖尿病肾病患者肾组织STOML2的表达及定位,采用慢病毒转染方法建立稳定过表达STOML2的HK-2细胞系,并应用Western blot检测肾小管上皮细胞钙粘蛋白(E-cadherin)、Fibronectin和STOML2的表达。结果 STOML2在DN患者肾组织的肾小管上皮细胞胞浆中表达明显升高。在高糖刺激HK-2细胞建立的EMT模型中,STOML2呈时间依赖表达上调。STOML2稳定高表达时,E-cadherin表达下调,而Fibronectin明显上调,即能促进肾小管上皮细胞发生EMT。结论 STOML2可能通过促进肾小管上皮细胞向间充质细胞分化,进而参与糖尿病肾病肾脏纤维化的发生发展。

Objective To investigate the expression and role of STOML2 in renal tissue of patients with type 2 diabetic nephropathy. Methods To detect he expression and localization of STOML2 in clinical renal tissue in patients with type 2 diabetic nephropathy by immunohistochemistry, and use lentiviral transfection method to establish a stable cell line of over-expressing STOML2, lastly apply western blot to detect the expression of E-cadherin, Fibronectin and STOML2 in renal tubular epithelial cells. Results STOML2 was significantly increased in renal tubular epithelial cytoplasm of patients with DN. In the EMT model of HK-2 cells stimulated by high glucose, STOML2 was increased in a time dependent. Overexpression of STOML2 led to E-cadherin down-regulated, while Fibronectin up-regulated, which promoted the occurrence of EMT in renal tubular epithelial cells. Conclusion STOML2 may be involved in the development and progression of renal fibrosis in diabetic nephropathy by mediating epithelial- mesenchymal transition of renal tubular epithelial cells.