| miR-34a通过靶向抑制Notch信号通路减轻糖尿病肾病小鼠的足细胞损伤 |
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| 引用本文： | 王欢岚,刘 红,张燕敏,陈伟栋. miR-34a通过靶向抑制Notch信号通路减轻糖尿病肾病小鼠的足细胞损伤[J]. 南方医科大学学报, 2022, 42(12): 1839-1845. DOI: 10.12122/j.issn.1673-4254.2022.12.12 |
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| 作者姓名： | 王欢岚 刘 红 张燕敏 陈伟栋 |
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| 作者单位： | 武汉市第一医院（武汉市中西医结合医院）肾内科，湖北 武汉 430022 |
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| 摘 要： | 目的 探讨miR-34a介导Notch信号通路对糖尿病肾病（DN）足细胞损伤及凋亡的影响。方法 体外实验：通过RT-PCR法检测高糖（30 mmol/L）环境下足细胞中miR-34a表达水平，构建miR-34a过表达足细胞系（miR-34a组）及其阴性对照（miR-NC组），使用荧光素酶实验验证miR-34a与Notch 1的靶向关系，构建Notch 1过表达足细胞系（Notch 1组）和miR-34a、Notch 1均表达升高细胞系（miR-34+Notch 1组）；采用CCK-8法检测足细胞存活情况，流式细胞法检测细胞凋亡情况，Western blot法检测细胞凋亡关蛋白水平。体内实验：通过高脂饮食和链脲佐菌素建立DN小鼠模型并分为模型组、miR-34a组（n=15/组），另选15只不做干预小鼠为对照组，miR-34a组和模型组小鼠分别尾静脉注射agomir-34a[80 mg/(kg·d)]、agomir-NC[80 mg/(kg·d)]，连续注射3 d,4周后，HE染色、TUNEL法分别观察肾组织病理、凋亡情况，Western blot检测肾组织凋亡相关蛋白和Notch 1蛋白...
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| 关 键 词： | 糖尿病肾病 miR-34a Notch信号通路 足细胞 凋亡 |
MiR-34a alleviates podocyte injury in mice with diabetic nephropathy by targeted downregulation of Notch signaling pathway |
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| WANG Huanlan,LIU Hong,ZHANG Yanmin,CHEN Weidong. MiR-34a alleviates podocyte injury in mice with diabetic nephropathy by targeted downregulation of Notch signaling pathway[J]. Journal of Southern Medical University, 2022, 42(12): 1839-1845. DOI: 10.12122/j.issn.1673-4254.2022.12.12 |
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| Authors: | WANG Huanlan LIU Hong ZHANG Yanmin CHEN Weidong |
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| Affiliation: | Department of Nephrology, Wuhan First Hospital (Wuhan Hospital of Integrated Traditional Chinese and Western Medicine), Wuhan 430022, China |
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| Abstract: | Objective To explore the effects of miR-34a on injury and apoptosis of podocytes in diabetic nephropathy (DN) and the role of Notch signaling pathway in mediating its effects. Methods The expression of miR-34a in podocytes exposed to high glucose (30 mmol/L) was detected using RT-PCR. A podocyte line with miR-34a overexpression was constructed, and the miRNA-target relationship between miR-34a and Notch 1 was verified with luciferase assay. The effects of overexpression of Notch 1 and both miR-34a and Notch 1 on podocyte survival and apoptosis were evaluated using CCK-8 and flow cytometry and by detecting apoptosis- related proteins using Western blotting. In a DN mouse model established by high- fat diet and streptozotocin, the effect of tail vein injection of agomir-34a and agomir-NC on pathology and apoptosis in the renal tissues were observed with HE staining and TUNEL staining, and the renal expressions of apoptosis-related proteins and Notch 1 protein were detected with Western blotting. Results High glucose exposure significantly lowered miR-34a expression in cultured human podocytes (P<0.05). The expression of Notch 1 was significantly lowered in miR-34a-overexpressing podocytes as compared with the cells with miR-NC transfection (P<0.05). Luciferase assay confirmed the mRNA-target relationship between miR-34a and Notch 1 (P<0.05). MiR-34a overexpression obviously promoted podocyte survival (P<0.05), reduced Notch 1 expression, and lowered apoptosis rate and the protein expressions of caspase-3, caspase- 9 and Bax/Bcl-2 levels in the cells (P<0.05), while the reverse changes were observed in Notch 1-overexpressing podocytes (P<0.05). In DN mouse models, treatment with miR-34a obviously alleviated renal pathologies. Compared with that in the control group, the expression level of miR-34a in the renal tissues was significantly lowered in DN model group (P<0.05) and increased in miR-34a group (P<0.05). The mice in the model group showed significantly higher apoptosis index of the renal tissues with increased expressions of caspase-3, caspase-9 and Notch 1 (P<0.05), which were lowered by treatment with miR-34a (P<0.05). Conclusion MiR-34a is capable of improving podocyte injury and apoptosis in DN mice by targeted downregulation of Notch 1. |
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| Keywords: | diabetic nephropathy miR-34a notch signaling pathway podocyte apoptosis, |
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