微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡

doi:10.16098/j.issn.0529-1356.2021.05.009

解剖学报 ›› 2021, Vol. 52 ›› Issue (5): 728-736.doi: 10.16098/j.issn.0529-1356.2021.05.009

微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡

高飞¹张欣欣¹ 杨冰¹ 陈素枝² 杨凤文² 檀淼³ 檀金川^2*

1.河北中医学院研究生学院，石家庄 050200; 2.河北省中医院肾病科，石家庄 050011; 3.河北医科大学第四医院内分泌科，石家庄 050011

收稿日期:2020-08-03 修回日期:2020-11-01 出版日期:2021-10-06 发布日期:2021-10-06
通讯作者: 檀金川 E-mail:1955981973@qq.com
基金资助:
河北省自然科学基金

MicroRNA-193a regulating Wilms’tumor gene 1 to promote mouse podocyte apoptosis in diabetic nephropathy#br#

GAO Fei¹ ZHANG Xin-xin¹ YANG Bing¹ CHEN Su-zhi² YANG Feng-wen² TAN Miao³ TAN Jin-chuan^2*#br#

1.Graduate Academy, Hebei University of Chinese Medicine, Shijiazhuang 050200, China; 2.Nephropathy of Traditional Chinese Medicine Hospital of Hebei Provincial, Shijiazhuang 050011, China; 3.Department of Endocrinology, the Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China

Received:2020-08-03 Revised:2020-11-01 Online:2021-10-06 Published:2021-10-06
Contact: TAN Jin-chuan E-mail:1955981973@qq.com

摘要/Abstract

摘要：

目的探讨微小RNA（miR）-193a对糖尿病肾病（DN）足细胞凋亡的影响及作用机制。方法通过体外高糖培养足细胞和体内小鼠腹腔注射链脲佐菌素(STZ)复制DN模型，将细胞或60只小鼠随机分为空白对照组、模型对照组、miR-193a抑制阴性对照组、miR-193a抑制组、miR-193a过表达阴性对照组和miR-193a过表达组。使用流式细胞术、免疫组织化学、免疫荧光、TUNEL、Real-time PCR和Western blotting检测DN小鼠和小鼠足细胞凋亡情况。结果 DN小鼠和高糖诱导的小鼠足细胞中Nephrin、Podocin表达减弱，细胞凋亡率显著升高，miR-193a高表达，小鼠肾脏和足细胞中cleaved-Caspase-3和Bax蛋白水平显著升高，Bcl-2蛋白水平显著下降，而miR-193a 抑制剂(inhibitor)可改善这一过程。DN小鼠和高糖培养的小鼠足细胞中Wilms瘤基因1（WT1） mRNA和蛋白表达水平显著降低，miR-193a inhibitor干预后WT1蛋白表达显著升高。上调WT1可降低miR-193a对高糖诱导的小鼠足细胞凋亡的影响。双荧光素酶报告实验证实了miR-193a和WT1之间的靶向关系。
结论 MiR-193a下调WT1的表达，促进DN足细胞凋亡。

关键词: 微小RNA-193a, Wilms瘤基因1, 高糖, 足细胞, 流式细胞术, 小鼠

Abstract:

Objective To explore the effect of microRNA(miR)-193a on the apoptosis of mouse podocytes in diabetic nephropathy (DN) and its mechanism. Methods The DN model was replicated by culturing podocytes with high glucose in vitro and intraperitoneal injection of streptozocin (STZ) in mice in vivo The cells or 60 mice were randomly divided into normal control(NC) group, model control group, and miR-NC inhibitor group, miR-193a inhibitor group, miR-NC mimic group and miR-193a mimic group. Flow cytometry, immunohistochemistry, TUNEL, Real-time PCR, Western blotting were used to examine the apoptosis of DN mice and mouse podocytes. Results The expression of Nephrin and Podocin in podocytes was weakened in DN mice and renal podocytes induced by high glucose, the apoptotic rate increased significantly, miR-193a was highly expressed, the levels of cleaved-Caspase-3 and Bax protein increased significantly, the level of Bcl-2 protein decreased significantly, and miR-193a inhibitor could improve this process. Wilms’tumor gene 1(WT1) mRNA and protein expression levels were significantly reduced in DN mice and podocytes cultured with high glucose. WT1 protein expression increased significantly after miR-193a inhibitor intervention, and WT1 protein expression was significantly reduced after miR-193a mimic transfection. Up-regulating WT1 could reduce the effect of miR-193a on the apoptosis of mouse podocytes induced by high glucose. The dual luciferase reporter experiment confirmed the targeting relationship between miR-193a and WT1. Conclusion MiR-193a down-regulates the expression of WT1 and promotes apoptosis of DN podocytes.

Key words: MicroRNA-193a, Wilms’tumor gene 1, High-glucose, Podocyte, Flow cytometry, Mouse

中图分类号:

R587.2

高飞张欣欣杨冰陈素枝杨凤文檀淼檀金川. 微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡[J]. 解剖学报, 2021, 52(5): 728-736.

GAO Fei ZHANG Xin-xin YANG Bing CHEN Su-zhi YANG Feng-wen TAN Miao TAN Jin-chuan. MicroRNA-193a regulating Wilms’tumor gene 1 to promote mouse podocyte apoptosis in diabetic nephropathy#br#[J]. Acta Anatomica Sinica, 2021, 52(5): 728-736.

参考文献

［1］ Haoran D, Qingquan L, Baoli L. Research progress on mechanism of podocyte depletion in diabetic nephropathy［J］. J Diabetes Res, 2017,16(7):1-10.
［2］ Sankrityayan H, Kulkarni YA, Gaikwad AB. Diabetic nephropathy: the regulatory interplay between epigenetics and microRNAs［J］. Pharmacol Res,2019,141(141):574-585.
［3］ Jiao TT, Han QX, Zhang D, et al. Research progress in the mechanism of microRNA in diabetic nephropathy［J］. Chinese Electronic Journal of Nephrology Research, 2019, 8(3):138-141. (in Chinese)
焦婷婷, 韩秋霞, 张冬,等. 微小RNA在糖尿病肾病中的作用机制研究进展［J］. 中华肾病研究电子杂志, 2019, 8(3):138-141. 
［4］ Ling Z, Wang X, Tao T, et al. Involvement of aberrantly activated HOTAIR/EZH2/miR-193a feedback loop in progression of prostate cancer［J］. J Exp Clin Cancer Res,2017,36(1):159-164.
［5］ Khordadmehr M, Shahbazi R, Sadreddini S, et al. miR-193: A new weapon against cancer［J］. J Cell Physiol, 2019,16(11):1-12.
［6］ Zhu YN, Ao Y, Li B, et al. Developmental disorder of podocytes and the related renal diseases［J］. Hereditas, 2018, 40(2): 116-125.
［7］ Liu XCh, Wang GF, Zhang ShF. Aerobic exercise can improve kidney oxidative stress damage in diabetic nephropathy model mice［J］. Chinese Tissue Engineering Research,2020,24(17):2712-2717. (in Chinese)
刘晓晨,王改凤,张社峰.有氧运动可改善糖尿病肾病模型小鼠肾脏的氧化应激损伤［J］.中国组织工程研究,2020,24(17):2712-2717. 
［8］ Jessica L. Harding1, Meda E, et al. Global trends in diabetes complications: a review of current evidence ［J］. Diabetologia, 2019,62(62):3-16.
［9］ Liang C, Liisa H, Changliang W, et al. Trends in the development of miRNA bioinformatics tools［J］. Brief Bioinform, 2019,20(5):1836-1852.
［10］ Chen C, Cheng MCh. The progress of exosomes and their applications in renal diseases［J］.Journal of Central South University (Medical Edition),2020,45(4):440-448. (in Chinese)
陈灿,成梅初.外泌体及其在肾脏疾病中的应用进展［J］.中南大学学报(医学版),2020,45(4):440-448. 
［11］ Zhang Y, Ding T, Tang DX, et al. Effects of tripterygium glycosides on autophagy in renal cells of diabetic nephropathy rats and related mechanisms［J］. Acta Anatomica Sinica,2020,51(3):373-377. (in Chinese)
张筠,丁婷,唐东兴,等.雷公藤多苷对糖尿病肾病大鼠肾脏细胞自噬的影响及相关机制［J］.解剖学报,2020,51(3):373-377. 
［12］ Zheng W, Pan ShK, Liu DW, et al. Progress in the treatment of diabetic nephropathy［J］. Chinese Journal of Nephrology, 2020, 36 (6): 476-480. (in Chinese)
郑文,潘少康,刘东伟,等.糖尿病肾病治疗进展［J］.中华肾脏病杂志,2020,36(6):476-480. 
［13］ Zhang W, Ren Y, Li J. Application of miR-193a/WT1/PODXL axis to estimate risk and prognosis of idiopathic membranous nephropathy［J］. Renal Failure, 2019, 41(1):704-717.
［14］ Zhibin H, Yong Z, Jianhua Z, et al. Urinary exosomal miR-193a can be a potential biomarker for the diagnosis of primary focal segmental glomerulosclerosis in children［J］. Biomed Res Int, 2017, 1(29):1-6.
［15］ Zapata-Benavides P, Arellano-Rodríguez M, Bollain-y-Goytia JJ, et al. Cytoplasmic localization of WT1 and decrease of miRNA-16-1 in nephrotic syndrome［J］. Biomed Res Int, 2017, 2(19):1-8.
［16］ Miyauchi M, Toyoda M, Kobayashi K, et al. Hypertrophy and loss of podocytes in diabetic nephropathy［J］. Internal Med,2009,48(18):1615-1620.
［17］ Shao ZhX, Xie JJ, Xu ZhH, et al. MicroRNA-21 transfection mediated changes in the biological behavior and radiosensitivity of esophageal squamous cell carcinoma cells［J］. Acta Anatomica Sinica, 2020, 51(3): 385-391. (in Chinese)
邵志雄,谢俊杰,徐振华,等.微小RNA-21转染介导食管鳞癌细胞生物学行为及细胞放射敏感性的变化［J］.解剖学报,2020,51(3):385-391. 
［18］ Wang S, Diao YJ, Zhu BB. MiR-193a-5p suppresses cell proliferation and induces cell apoptosis by regulating HOXA7 in human ovarian cancer［J］. Neoplasma, 2020, 67(4):825-833.
［19］ Wan J, Hou X, Zhou Z, et al. WT1 ameliorates podocyte injury via repression of EZH2/β-catenin pathway in diabetic nephropathy［J］. Free Radic Biol Med,2017(108):280-299.
［20］ Li J, Chen Y, Shen L, et al. Improvement of membranous nephropathy by inhibition of miR-193a to affect podocytosis via targeting WT1［J］. J Cell Biochem,2019,120(3):3438-3446.
［21］ Wu C, Wang S, Xu C, et al. WT1 enhances proliferation and impedes apoptosis in KRAS mutant NSCLC via targeting cMyc［J］. Cell Physiol Biochem,2015,35(2):647-662.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[3]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[4]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[5]	赵卫明李晓余国营王改平常翠芳 . 丝氨酸肽酶抑制因子Kazal型1对肝癌细胞增殖的影响及其机制 [J]. 解剖学报, 2023, 54(6): 695-702.
[6]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.
[7]	陈子璇司丽娜舒薇菡张欣魏晨阳程露阳杨松鹤乔跃兵. 褪黑素调控下丘脑PI3K/Akt/mTOR信号延缓雌性小鼠青春期启动的机制 [J]. 解剖学报, 2023, 54(6): 644-651.
[8]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.
[9]	刘叶楚亚楠徐岑璐何佳澄苏炳银太颢然. Roscovitine挽救帕金森病小鼠相关脑区神经元丢失和神经炎症[J]. 解剖学报, 2023, 54(6): 635-643.
[10]	魏纯纯王平林方兴麻献华章卫平谢志芳 . 小鼠棕色脂肪透射电子显微镜样品固定方法的改良 [J]. 解剖学报, 2023, 54(6): 738-743.
[11]	阎锟武筱林刘英娟葛科立任雷鸣李红云 . 脑蛋白水解物-Ⅰ对帕金森病小鼠肠道菌群的调节作用[J]. 解剖学报, 2023, 54(5): 497-504.
[12]	顾玲张萍宋科昕李娜陈虹宇张婕丛敬翟效月 . 发生发育小鼠肾输尿管芽分支的形态特点[J]. 解剖学报, 2023, 54(5): 593-598.
[13]	何可可李远迪文廷浩朱婕高杰胡蓉韩锋苏敏. 红细胞膜相关蛋白通过IL-6/STAT3/ROR-γt 信号通路抑制小鼠实验性自身免疫性脑脊髓炎中Th17细胞分化[J]. 解剖学报, 2023, 54(5): 538-545.
[14]	范文娟陈旭东陈永芳杨旭光金少举赵志军邓锦波. 体外培养大脑皮质类器官与活体大脑皮质发育的比较[J]. 解剖学报, 2023, 54(4): 383-391.
[15]	张沂洲李莎米世雄左洪春张倩倩李晗琳雷梓晗张东泽崔慧先. 雌二醇经雌激素受体调控分拣蛋白相关受体A对小鼠海马神经元树突棘密度和突触蛋白表达的影响[J]. 解剖学报, 2023, 54(3): 261-268.

微小RNA-193a调控Wilms瘤基因1促进小鼠糖尿病肾病足细胞凋亡

MicroRNA-193a regulating Wilms’tumor gene 1 to promote mouse podocyte apoptosis in diabetic nephropathy#br#

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价