NUP160 knockdown inhibits the progression of diabetic nephropathy in vitro and in vivo

Jiayong Xie; Zhi Chen; Gang Yao; Ying Yuan; Wenjuan Yu; Qiang Zhu

doi:10.1016/j.reth.2022.05.011

NUP160 knockdown inhibits the progression of diabetic nephropathy in vitro and in vivo

Regen Ther. 2022 Jun 17:21:87-95. doi: 10.1016/j.reth.2022.05.011. eCollection 2022 Dec.

Authors

Jiayong Xie^{1

2}, Zhi Chen³, Gang Yao⁴, Ying Yuan¹, Wenjuan Yu⁴, Qiang Zhu¹

Affiliations

¹ Department of Nephrology, Xinghua People's Hospital, Taizhou 225700, Jiangsu, China.
² Kangda College of Nanjing Medical University, Lianyungang 222000, Jiangsu, China.
³ Department of Laboratory, Xinghua People's Hospital, Taizhou 225700, Jiangsu, China.
⁴ Department of Nephrology, Second Affiliated Hospital of Nanjing Medical University, Nanjing 210011, Jiangsu, China.

Abstract

Diabetic nephropathy (DN) is a severe diabetic complication and podocyte damage is a hallmark of DN. The Nucleoporin 160 (NUP160) gene was demonstrated to regulate cell proliferation and apoptosis in mouse podocytes. This study explored the possible role and mechanisms of NUP160 in high glucose-triggered podocyte injury. A rat model of DN was established by intraperitoneal injection of 60 mg/kg streptozotocin (STZ). Podocytes were treated with 33 mM high glucose. The effects of the Nup160 on DN and its mechanisms were assessed using MTT, flow cytometry, Western blot, ELISA, RT-qPCR, and luciferase reporter assays. The in vivo effects of NUP160 were analyzed by HE, PAS, and MASSON staining assays. The NUP160 level was significantly upregulated in podocytes treated with 33 mM high glucose. Functionally, NUP160 knockdown alleviated high glucose-induced apoptosis and inflammation in podocytes. Mechanistically, miR-495-3p directly targeted NUP160, and lncRNA HCG18 upregulated NUP160 by sponging miR-495-3p by acting as a ceRNA. Additionally, NUP160 overexpression reversed the effects of HCG18 knockdown in high glucose treated-podocytes. The in vivo assays indicated that NUP160 knockdown alleviated the symptoms of DN rats. NUP160 knockdown plays a key role in preventing the progression of DN, suggesting that targeting NUP160 may be a potential therapeutic strategy for DN treatment.

Keywords: DN, diabetic nephropathy; Diabetic nephropathy; ECM, extracellular matrix; ELISA, Enzyme-linked immunosorbent assay; HCG18; HE, Hematoxylin-eosin; NPC, nuclear pore complex; NUP160; NUP160, Nucleoporin 160; Podocytes; STZ, streptozotocin; ceRNAs, competitive endogenous RNAs; miRNAs, MicroRNAs.