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

GAO Fei; WANG Ze-Ze; YANG Bing; TAN Jin-Chuan

doi:10.16098/j.issn.0529-1356.2021.05.009

PDF(26635 KB)

Acta Anatomica Sinica ›› 2021, Vol. 52 ›› Issue (5) : 728-736. DOI: 10.16098/j.issn.0529-1356.2021.05.009

Cell and Molecules Biology

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#

Author information +

History +

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

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

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 https://doi.org/10.16098/j.issn.0529-1356.2021.05.009

References

［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.