硫酸吲哚酚促进肾纤维化的机制研究  

Indoxyl sulfate induces renal fibrosis in mice

作　　者：余彦霖[1] 肖堂利[1] 刘亮[1] 何婷[1] 徐新丽[1] 管旭[1] 杨可[1] 赵景宏[1] 

机构地区：[1]第三军医大学新桥医院肾内科,全军肾脏病中心,重庆400037

出　　处：《第三军医大学学报》2016年第2期119-123,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金面上项目(81270290)~~

摘　　要：目的探讨尿毒症毒素硫酸吲哚酚（indoxyl sulfate,IS）诱导肾纤维化的作用及可能机制。方法 24只3周龄健康雄性CD-1小鼠,体质量14-15 g,建立1/2肾切除模型后,采用完全随机分组法将小鼠分为IS处理组（n=12）和对照组（n=12）,分别给予小鼠腹腔注射IS 100 mg/（kg·d）和等体积灭菌磷酸缓冲溶液,IS注射4周后留取肾脏标本,Masson染色观察小鼠肾间质胶原沉积情况,免疫组化染色检测观察小鼠肾脏α-平滑肌肌动蛋白（α-SMA）和β-catenin蛋白表达变化,Western blot检测β-catenin蛋白表达变化;用不同浓度的IS（0、2、10、50 mg/L）孵育人近端肾小管上皮细胞（HK-2细胞）72 h后,Western blot检测β-catenin表达变化。结果与对照组比较,Masson染色发现腹腔注射IS 4周后,小鼠肾脏胶原沉积明显增加[（2.43±0.51）vs（6.57±0.51）,P〈0.05],IS处理组肾小管上皮细胞β-catenin[（0.87±0.74）vs（10.87±1.21）,P〈0.05]以及α-SMA[（1.30±0.56）vs（13.53±0.50）,P〈0.05]的表达均明显增加;与对照组相比,IS处理组HK-2细胞的β-catenin表达显著增加,且呈剂量依赖效应[（0.71±0.71）vs（0.96±0.70）或（0.71±0.71）vs（1.10±0.95）,P〈0.01]。结论 IS可能通过激活Wnt/β-catenin信号通路发挥促小鼠肾纤维化的作用。Objective To explore the effect of uremic toxin indoxyl sulfate（ IS） on inducing renal fibrosis in mice and its underlying mechanism. Methods A total of 24 healthy 3-week-old male CD-1 mice（ weighting 14 - 15 g） were used to establish 1 /2 nephrectomy model and then randomly divided into IS treatment group（ n = 12） and blank control group（ n = 12）. The IS treatment group received intraperitoneal injection of IS at dose of 100 mg /（ kg · d）,and the control group received the same volume of phosphate buffer saline for 4 weeks. At the end of 4 weeks,the kidney samples were taken,and the mouse renal interstitial collagen deposition was observed by Masson staining. Expression levels of renal α-smooth muscle actin（ α-SMA） and β-catenin were detected by immunohistochemical staining,and the changes of β-catenin protein were also detected by Western blotting. Human renal proximal tubular epithelial cells（ HK-2 cells）were incubated with different concentrations of IS（ 0,2,10,and 50 mg / L） for 72 h,and the expression ofβ-catenin was measured by Western blotting. Results Compared with the control group,Masson staining showed that the IS-injected mice had significantly increased collagen deposition（ 2. 43 ± 0. 51 vs 6. 57 ± 0. 51,P〈0. 05）,and the expression levels of β-catenin（ 0. 87 ± 0. 74 vs 10. 87 ± 1. 21,P〈0. 05） and α-SMA（ 1. 30 ± 0. 56 vs 13. 53 ± 0. 50,P〈0. 05） in the proximal and distal tubule epithelial cells of the IS-injected mice were obviously increased. In the HK-2 cells incubated with IS,the protein levels of β-catenin increased significantly（ 0. 71 ± 0. 71 vs 0. 96 ± 0. 70,0. 71 ± 0. 71 vs 1. 10 ± 0. 95,P〈0. 01） in a dose-dependent manner. Conclusion IS can promote the progress of renal fibrosis in the mice,which may be through activating Wnt / β-catenin signaling pathway.

关 键 词：硫酸吲哚酚 WNT/Β-CATENIN 肾纤维化 

分 类 号：R332[医药卫生—人体生理学]