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缺氧肾小管上皮细胞Rab7/Caveolin-1/MMP-2轴失调促进肾纤维化
作者：程正源1  张晓娟2  方桂琳2  许卿3  杨一琼3  陈平圣3 
单位：1. 东南大学医学院附属马鞍山市人民医院 内科, 安徽 马鞍山 243099;
2. 东部战区总医院 肾内科, 江苏 南京 210016;
3. 东南大学医学院 病理学系, 江苏 南京 210009
关键词：Ras相关GTP结合蛋白7 小窝蛋白-1 基质金属蛋白酶-2 肾纤维化 小鼠 
分类号：R692.9
出版年·卷·期（页码）：2023·42·第一期（57-64）
摘要：

目的：研究肾纤维化晚期Ras相关GTP结合蛋白7（Ras-related GTP binding protein 7，Rab7）可否通过小窝蛋白-1（Caveolin-1）的介导调节基质金属蛋白酶-2（matrix metalloproteinase-2，MMP-2）的活性，从而对肾纤维化产生影响。方法：通过单侧输尿管结扎（unilateral ureteral obstruction，UUO）法构建C57BL/6小鼠肾纤维化模型，检测肾纤维化组织中缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）、Rab7和Caveolin-1的表达以及MMP-2的活性。利用肾小管上皮细胞株（HK-2）进行体外实验，构建HK-2 Rab7-shRNA细胞，将HK-2和HK-2 Rab7-shRNA细胞分别进行常氧和缺氧培养，并检测各组中HIF-1α、Rab7、Caveolin-1的表达和MMP-2的活性，以及纤维化相关指标I型胶原（Collagen-1）、波形蛋白（Vimentin）和α-平滑肌肌动蛋白（α-smooth muscle actin，α-SMA）的表达。结果：利用小鼠UUO模型发现，相较正常肾脏组织，肾纤维化组织中Rab7和Caveolin-1的表达均上调，而MMP-2的活性反而下降。进一步体外实验显示，在HK-2细胞中Rab7下调后可通过抑制Caveolin-1的表达进而提高MMP-2的活性，并会使Collagen-1、Vimentin和α-SMA等反映纤维化的指标表达下调。结论：Rab7与MMP-2的活性密切相关；Rab7对MMP-2活性的调节通过Caveolin-1介导；当Rab7表达受抑后，可促进肾纤维化的缓解。

Objective: To study whether Ras-related GTP binding protein 7(Rab7) can regulate the activity of matrix metalloproteinase-2(MMP-2) through the mediation of Caveolin-1 in the advanced stage of renal fibrosis,thereby affecting renal fibrosis. Methods: The renal fibrosis model of C57BL/6 mice was established by unilateral ureteral obstruction(UUO) method, and the expressions of hypoxia inducible factor-1α(HIF-1α), Rab7, Caveolin-1 and the activity of MMP-2 were detected in renal fibrosis tissues.The renal tubular epithelial cell line(HK-2) was used for in vitro experiments. First, HK-2 Rab7-shRNA cells were constructed. HK-2 and HK-2 Rab7-shRNA cells were cultured in normoxia and hypoxia respectively, and HIF-1α, Rab7, Caveolin-1 expression and MMP-2 activity were detected in each group. Then the expression of fibrosis related indicators, such as Collagen-1, Vimentin and α-smooth muscle actin(α-SMA) were also detected. Results: Using the mouse UUO model, it was found that the expression of Rab7 and Caveolin-1 were both upregulated in renal fibrosis tissues compared with normal kidney tissues, while the activity of MMP-2 was decreased. Further in vitro experiments revealed that Rab7 down-regulation in HK-2 cells could increase the activity of MMP-2 in HK-2 supernatant by inhibiting Caveolin-1 expression,and down-regulate the expression of Collagen-1, Vimentin and α-SMA. Conclusion: Rab7 is closely related to MMP-2 activity,and the regulation of MMP-2 activity by Rab7 is mediated by Caveolin-1. When the expression of Rab7 is inhibited, it can promote the remission of renal fibrosis.
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