Department of Pathology, Affiliated Hospital of Jiangnan University, Wuxi, Jiangsu, China.
Department of Pathology, Zhongshan Hospital, Fudan University, Shanghai, China.
Braz J Med Biol Res. 2020 Apr 6;53(4):e9220. doi: 10.1590/1414-431X20209220. eCollection 2020.
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. In the early stage, our group studied the effects of Rab7 on production and degradation of extracellular matrix in hypoxic renal tubular epithelial cells. Because cell culture in vitro is different from the environment in vivo, it is urgent to understand the effects in vivo. In our current study, we established a renal fibrosis model in Rab7-knock-in mice (prepared by CRISPR/Cas9 technology) and wild type (WT) C57BL/6 mice using unilateral ureteral obstruction (UUO). Seven and 14 days after UUO, the expression of the Rab7 protein in WT mice, as well as the autophagic activity, renal function, and the degree of renal fibrosis in WT and Rab7-knock-in mice were examined by blood biochemical assay, hematoxylin-eosin and Masson staining, immunohistochemistry, and western blotting. We found that the Rab7 expression in WT mice increased over time. Furthermore, the autophagic activity constantly increased in both groups, although it was higher in the Rab7-knock-in mice than in the WT mice at the same time point. Seven days after UUO, the degree of renal fibrosis was milder in the Rab7-knock-in mice than in the WT mice, but it became more severe 14 days after surgery. Similar results were found for renal function. Therefore, Rab7 suppressed renal fibrosis in mice initially, but eventually it aggravated fibrosis with the activation of autophagy.
Rab7 是 Rab 家族的重要成员,与自噬、内吞作用、细胞凋亡和肿瘤抑制密切相关,但很少有研究描述其与肾纤维化的关系。在早期,我们的研究小组研究了 Rab7 对缺氧肾小管上皮细胞细胞外基质产生和降解的影响。由于体外细胞培养与体内环境不同,因此迫切需要了解体内的影响。在目前的研究中,我们使用单侧输尿管梗阻(UUO)在 Rab7 敲入小鼠(由 CRISPR/Cas9 技术制备)和野生型(WT)C57BL/6 小鼠中建立了肾纤维化模型。UUO 后 7 天和 14 天,通过血液生化检测、苏木精-伊红和 Masson 染色、免疫组织化学和 Western blot 检测 WT 小鼠 Rab7 蛋白的表达以及 WT 和 Rab7 敲入小鼠的自噬活性、肾功能和肾纤维化程度。我们发现 WT 小鼠的 Rab7 表达随时间增加。此外,两组的自噬活性持续增加,尽管在同一时间点 Rab7 敲入小鼠的自噬活性高于 WT 小鼠。UUO 后 7 天,Rab7 敲入小鼠的肾纤维化程度比 WT 小鼠轻,但手术后 14 天则更严重。肾功能也有类似结果。因此,Rab7 最初抑制了小鼠的肾纤维化,但随着自噬的激活,最终加重了纤维化。