The Effects Of Rab7 Gene Up-regulation On Renal Fibrosis Induced By Unilateral Ureteral Obstruction In Mice

Background: Rab7,an important member of the Rab family,is closely related to autophagy,endocytosis,apoptosis and tumor suppression but few studies have described its association with renal fibrosis.The effects of autophagy and endocytosis of hypoxic renal tubular epithelial cells on the production and degradation of extracellular matrix were studied in the early stage by our group,and it was found that Rab7 played an important role in this process.Because cell culture in vitro is different from the environment in vivo,it is urgent to understand the overall level.Objective: Rab7-knock-in mice and renal fibrosis models are constructed to detect Rab7 gene expression and related indicators of autophagy and fibrosis in renal tissues at different time points,so as to compare the differences between wild type(WT)mice and mice with overexpression of Rab7 gene and reveal the change rules and possible role of Rab7 gene in the development process of renal fibrosis.Methods: In our current study,a renal fibrosis model is established in Rab7-knock-in mice(prepared by CRISPR/Cas9 technology)and WT C57BL/6 mice using unilateral ureteral obstruction(UUO).7 and 14 days after the UUO,the expression of the Rab7 protein in WT mice,as well as the autophagic activity and the degree of renal fibrosis in WT mice and Rab7-knock-in mice are examined by HE staining,Masson staining,immunohistochemistry and western blotting.Results: The Rab7 expression in WT mice increase over time.Furthermore,the autophagic activity constantly increase in both groups,although it is higher in the Rab7-knock-in mice than in the WT mice at the same time point.7 days after the UUO,the degree of renal fibrosis is milder in the Rab7-knock-in mice than in the WT mice,but it becomes severer 14 days after surgery.Conclusion:(1)Rab7-knock-in mice are successfully established.(2)initially Rab7 suppresses renal fibrosis in mice,but eventually it aggravates fibrosis with the activation of autophagy.