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©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
MicroRNA-630 alleviates inflammatory reactions in rats with diabetic kidney disease by targeting toll-like receptor 4
Qi-Shun Wu, Dan-Na Zheng, Cheng Ji, Hui Qian, Juan Jin, Qiang He
Qi-Shun Wu, Dan-Na Zheng, Qiang He, Graduate School, Medical College of Soochow University, Suzhou 215006, Jiangsu Province, China
Qi-Shun Wu, Dan-Na Zheng, Qiang He, Department of Nephrology, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou 314408, Zhejiang Province, China
Qi-Shun Wu, Department of Nephrology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, Jiangsu Province, China
Dan-Na Zheng, Department of Nephrology, Zhejiang Provincial People’s Hospital, Affiliated People’s Hospital, Hangzhou Medical College, Hangzhou 314408, Zhejiang Province, China
Cheng Ji, Hui Qian, Molecular Inspection Laboratory, School of Medicine, Jiangsu University, Zhenjiang 212000, Jiangsu Province, China
Juan Jin, Qiang He, Department of Nephrology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Zhejiang Provincial Hospital of Traditional Chinese Medicine, Hangzhou 310060, Zhejiang Province, China
Author contributions: Wu QS conceived and designed the experiments. Wu QS and Zheng DN performed the experiments, analysed the data, and prepared all the figures; Ji C, Qian H, Jin J, and He Q provided technical support; Wu QS wrote the manuscript; and all authors contributed to the article and approved the submitted version.
Supported by the Huadong Medicine Joint Funds of the Zhejiang Provincial Natural Science Foundation of China, No. LHDMZ22H050001; the Construction of Key Projects by Zhejiang Provincial Ministry, No. WKJ-ZJ-2302; the Zhejiang Province Chinese Medicine Modernization Program, No. 2020ZX001; the Key Project of Scientific Research Foundation of Chinese Medicine, No. 2022ZZ002; the “Pioneer” and “LeadingGoose” R&D Program of Zhejiang, No. 2022C03118 and 2023C03075; and the Key Project of Basic Scientific Research Operating Funds of Hangzhou Medical College, No. KYZD202002.
Institutional review board statement: This study did not involve human experimentation.
Institutional animal care and use committee statement: The study was reviewed and approved by the Ethics Committee of the Laboratory Animal of Jiangsu University Institutional Review Board (Approval No. UJS-IACUC-AP-2022081615).
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Qiang He, MD, Chief Physician, Professor, Graduate School, Medical College of Soochow University, No. 1 Shizi Street, Gusu District, Suzhou 215006, Jiangsu Province, China.
strong_he@163.com
Received: August 31, 2023
Peer-review started: August 31, 2023
First decision: December 25, 2023
Revised: January 6, 2024
Accepted: January 29, 2024
Article in press: January 29, 2024
Published online: March 15, 2024
Processing time: 197 Days and 5.3 Hours
BACKGROUND
Diabetic kidney disease (DKD) is a major complication of diabetes mellitus. Renal tubular epithelial cell (TEC) damage, which is strongly associated with the inflammatory response and mesenchymal trans-differentiation, plays a significant role in DKD; However, the precise molecular mechanism is unknown. The recently identified microRNA-630 (miR-630) has been hypothesized to be closely associated with cell migration, apoptosis, and autophagy. However, the association between miR-630 and DKD and the underlying mechanism remain unknown.
AIM
To investigate how miR-630 affects TEC injury and the inflammatory response in DKD rats.
METHODS
Streptozotocin was administered to six-week-old male rats to create a hyperglycemic diabetic model. In the second week of modeling, the rats were divided into control, DKD, negative control of lentivirus, and miR-630 overexpression groups. After 8 wk, urine and blood samples were collected for the kidney injury assays, and renal tissues were removed for further molecular assays. The target gene for miR-630 was predicted using bioinformatics, and the association between miR-630 and toll-like receptor 4 (TLR4) was confirmed using in vitro investigations and double luciferase reporter gene assays. Overexpression of miR-630 in DKD rats led to changes in body weight, renal weight index, basic blood parameters and histopathological changes.
RESULTS
The expression level of miR-630 was reduced in the kidney tissue of rats with DKD (P < 0.05). The miR-630 and TLR4 expressions in rat renal TECs (NRK-52E) were measured using quantitative reverse transcription polymerase chain reaction. The mRNA expression level of miR-630 was significantly lower in the high-glucose (HG) and HG + mimic negative control (NC) groups than in the normal glucose (NG) group (P < 0.05). In contrast, the mRNA expression level of TLR4 was significantly higher in these groups (P < 0.05). However, miR-630 mRNA expression increased and TLR4 mRNA expression significantly decreased in the HG + miR-630 mimic group than in the HG + mimic NC group (P < 0.05). Furthermore, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), and IL-6 were significantly higher in the HG and HG + mimic NC groups than in NG group (P < 0.05). However, the levels of these cytokines were significantly lower in the HG + miR-630 mimic group than in the HG + mimic NC group (P < 0.05). Notably, changes in protein expression were observed. The HG and HG + mimic NC groups showed a significant decrease in E-cadherin protein expression, whereas TLR4, α-smooth muscle actin (SMA), and collagen IV protein expression increased (P < 0.05). Conversely, the HG + miR-630 mimic group exhibited a significant increase in E-cadherin protein expression and a notable decrease in TLR4, α-SMA, and collagen IV protein expression than in the HG + mimic NC group (P < 0.05). The miR-630 targets TLR4 gene expression. In vivo experiments demonstrated that DKD rats treated with miR-630 agomir exhibited significantly higher miR-630 mRNA expression than DKD rats injected with agomir NC. Additionally, rats treated with miR-630 agomir showed significant reductions in urinary albumin, blood glucose, TLR4, and proinflammatory markers (TNF-α, IL-1β, and IL-6) expression levels (P < 0.05). Moreover, these rats exhibited fewer kidney lesions and reduced infiltration of inflammatory cells.
CONCLUSION
MiR-630 may inhibit the inflammatory reaction of DKD by targeting TLR4, and has a protective effect on DKD.
Core Tip: This study revealed that microRNA-630 (miR-630) expression in the renal tissue was significantly lower in diabetic kidney disease (DKD) rats than in normal rats. The miR-630 alleviates renal injury and inflammatory reactions in DKD rats by targeting toll-like receptor 4. Our findings provide new insights into the pathogenesis of DKD indicating that miR-630 may be a potential noninvasive biomarker for diagnosing and predicting the prognosis of DKD.
