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Journal of Traditional Chinese Medicine >

2024 , Vol. 44 >Issue 1: 44 - 53

DOI: https://doi.org/10.19852/j.cnki.jtcm.20231204.004

Original articles

Neferine inhibits the progression of diabetic nephropathy by modulating the miR-17-5p/nuclear factor E2-related factor 2 axis

  • Hongmei HUANG ,
  • Maojun YANG ,
  • Ting LI ,
  • Dandan WANG ,
  • Ying LI ,
  • Xiaochi TANG ,
  • Lu YUAN ,
  • Shi GU ,
  • Yong XU
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  • 1 Department of Endocrinology, Chengdu Shuangliu District First People's Hospital (West China Airport of Sichuan University), Chengdu 610200, China
    2 Department of Endocrinology, Affiliated Hospital of Southwest Medical University, Luzhou 646000, China

Received date: 2023-12-12

  Accepted date: 2023-03-17

  Online published: 2023-12-04

Supported by

Chengdu Health and Wellness Commission: Exploring the Mechanism of Neferine on Diabetic Nephropathy Based on miR-17/Nrf2 Axis(2021127)

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Abstract

OBJECTIVE: To investigate the effect of Neferine (Nef) on diabetic nephropathy (DN) and to explore the mechanism of Nef in DN based on miRNA regulation theory.

METHODS: A DN mouse model was constructed and treated with Nef. Serum creatinine (Crea), blood urea (UREA) and urinary albumin were measured in mice by kits, and renal histopathological changes and fibrosis were observed by hematoxylin-eosin staining and Masson staining. Renal tissue superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) activities were measured by enzyme-linked immunosorbent assay (ELISA). Western blotting was used to detect the expression of nuclear factor E2-related factor 2 (Nrf2)/ heme oxygenase 1 (HO-1) signaling pathway-related proteins in kidney tissues. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to detect the expression of miR-17-5p in kidney tissues. Subsequently, a DN in vitro model was constructed by high glucose culture of human mesangial cells (HMCs), cells were transfected with miR-17-5p mimic and/or treated with Nef, and we used qRT-PCR to detect cellular miR-17 expression, flow cytometry to detect apoptosis, ELISAs to detect cellular SOD, MDA, and GSH-Px activities, Western blots to detect Nrf2/HO-1 signaling pathway-related protein expression, and dual luciferase reporter gene assays to verify the targeting relationship between Nrf2 and miR-17-5p.

RESULTS: Administration of Nef significantly reduced the levels of blood glucose, Crea, and UREA and the expression of miR-17-5p, improved renal histopathology and fibrosis, significantly reduced MDA levels, elevated SOD and GSH-Px activities, and activated Nrf2 expression in kidney tissues from mice with DN. Nrf2 is a post-transcriptional target of miR-17-5p. In HMCs transfected with miR-17-5p mimics, the mRNA and protein levels of Nrf2 were significantly suppressed. Furthermore, miR-17-5p overexpression and Nef intervention resulted in a significant increase in high glucose-induced apoptosis and MDA levels in HMCs and a significant decrease in the protein expression of HO-1 and Nrf2.

CONCLUSION: Collectively, these results indicate that Nef has an ameliorative effect on DN, and the mechanism may be through the miR-17-5p/Nrf2 pathway.

Key words： diabetic nephropathies; neferine; miR-17-5p; NF-E2-related factor 2; oxidative stress

Cite this article

Hongmei HUANG , Maojun YANG , Ting LI , Dandan WANG , Ying LI , Xiaochi TANG , Lu YUAN , Shi GU , Yong XU . Neferine inhibits the progression of diabetic nephropathy by modulating the miR-17-5p/nuclear factor E2-related factor 2 axis[J]. Journal of Traditional Chinese Medicine, 2024 , 44(1) : 44 -53 . DOI: 10.19852/j.cnki.jtcm.20231204.004

References

1. Samsu N. Diabetic nephropathy: challenges in pathogenesis, diagnosis, and treatment. Biomed Res Int 2021; 2021: 1497449.
2. Dai X, Liao R, Liu C, et al. Epigenetic regulation of TXNIP-mediated oxidative stress and NLRP 3 inflammasome activation contributes to SAHH inhibition-aggravated diabetic nephropathy. Redox Biol 2021; 45: 102033.
3. Gerardo Yanowsky-Escatell F, Andrade-Sierra J, Pazarín-Villase?or L, et al. The role of dietary antioxidants on oxidative stress in diabetic nephropathy. Iran J Kidney Dis 2020; 14: 81-94.
4. Sagoo MK, Gnudi L. Diabetic nephropathy: is there a role for oxidative stress? Free Radic Biol Med 2018; 116: 50-63.
5. Tang YS, Zhao YH, Zhong Y, et al. Neferine inhibits LPS-ATP-induced endothelial cell pyroptosis via regulation of ROS/NLRP3/Caspase-1 signaling pathway. Inflamm Res 2019; 68: 727-38.
6. Bharathi Priya L, Huang CY. An updated review on pharmacological properties of neferine-A bisbenzylisoquinoline alkaloid from Nelumbo nucifera. J Food Biochem 2021; 45: e13986.
7. Wang Y, Wang S, Wang R, et al. Neferine exerts antioxidant and anti-inflammatory effects on carbon tetrachloride-induced liver fibrosis by inhibiting the MAPK and NF-κB/IκBα pathways. Evid Based Complement Alternat Med 2021; 2021: 4136019.
8. Li S, Zhang Y, Zhang J, et al. Neferine exerts ferroptosis-inducing effect and antitumor effect on thyroid cancer through Nrf2/HO-1/NQO1 inhibition. J Oncol 2022; 2022: 7933775.
9. Liu Y, Wang J, Zhang X, Wang L, Hao T, Cheng Y. Scutellarin exerts hypoglycemic and renal protective effects in db/db mice via the Nrf2/HO-1 signaling pathway. Oxid Med Cell Longev 2019; 2019: 1354345.
10. Xing L, Guo H, Meng S, et al. Klotho ameliorates diabetic nephropathy by activating Nrf2 signaling pathway in podocytes. Biochem Biophys Res Commun 2021; 534: 450-56.
11. He M, Wang J, Yin Z, et al. MiR-320a induces diabetic nephropathy via inhibiting MafB. Aging (Albany NY) 2019; 11: 3055-79.
12. Guo Y, Du F, Tan YL, Luo J, Xiong D, Song WT. VEGF-mediated angiogenesis in retinopathy of prematurity is co-regulated by miR-17-5p and miR-20a-5p. Biochem Cell Biol 2021; 99: 414-23.
13. Wei J, Hao Q, Chen C, et al. Epigenetic repression of miR-17 contributed to di (2-ethylhexyl) phthalate-triggered insulin resistance by targeting Keap1-Nrf2/miR-200a axis in skeletal muscle. Theranostics 2020; 10: 9230-48.
14. Du KY, Qadir J, Yang BB, Yee AJ, Yang W. Tracking miR-17-5p levels following expression of seven reported target mRNAs. Cancers (Basel) 2022; 14: 2585.
15. Cai TT, Ye XL, Li RR, et al. Resveratrol modulates the gut microbiota and inflammation to protect against diabetic nephropathy in mice. Front Pharmacol 2020; 11: 1249.
16. Wu F, Ji A, Zhang Z, Li J, Li P. miR-491-5p inhibits the proliferation and migration of A549 cells by FOXP4. Exp Ther Med 2021; 21: 622.
17. Duan YR, Chen BP, Chen F, et al. LncRNA lnc-ISG20 promotes renal fibrosis in diabetic nephropathy by inducing AKT phosphorylation through miR-486-5p/NFAT5. J Cell Mol Med 2021; 25: 4922-37.
18. Shen Y, Chen W, Han L, et al. VEGF-B antibody and interleukin-22 fusion protein ameliorates diabetic nephropathy through inhibiting lipid accumulation and inflammatory responses. Acta Pharm Sin B 2021; 11: 127-42.
19. Lee SY, An HJ, Kim JM, et al. PINK1 deficiency impairs osteoblast differentiation through aberrant mitochondrial homeostasis. Stem Cell Res Ther 2021; 12: 589.
20. Ma H, Bell KN, Loker RN. qPCR and qRT-PCR analysis: Regulatory points to consider when conducting biodistribution and vector shedding studies. Mol Ther Methods Clin Dev 2021; 20: 152-68.
21. Pillai-Kastoori L, Schutz-Geschwender AR, Harford JA. A systematic approach to quantitative Western blot analysis. Anal Biochem 2020; 593: 113608.
22. Xian Y, Gao Y, Lü W, et al. Resveratrol prevents diabetic nephropathy by reducing chronic inflammation and improving the blood glucose memory effect in non-obese diabetic mice. Naunyn Schmiedebergs Arch Pharmacol 2020; 393: 2009-17.
23. Zheng C, Huang L, Luo W, et al. Inhibition of STAT3 in tubular epithelial cells prevents kidney fibrosis and nephropathy in STZ-induced diabetic mice. Cell Death Dis 2019; 10: 848.
24. Bai L, Li A, Gong C, Ning X, Wang Z. Protective effect of rutin against bleomycin induced lung fibrosis: involvement of TGF-β1/α-SMA/Col Ⅰ and Ⅲ pathway. Biofactors 2020; 46: 637-44.
25. Huang W, Man Y. Short-chain fatty acids ameliorate diabetic nephropathy via GPR43-mediated inhibition of oxidative stress and NF-κB signaling. Oxid Med Cell Longev 2020; 2020: 4074832.
26. Qiao S, Liu R, Lv C, et al. Bergenin impedes the generation of extracellular matrix in glomerular mesangial cells and ameliorates diabetic nephropathy in mice by inhibiting oxidative stress via the mTOR/β-TrcP/Nrf2 pathway. Free Radic Biol Med 2019; 145: 118-35.
27. Bellezza I, Giambanco I, Minelli A, Donato R. Nrf2-Keap1 signaling in oxidative and reductive stress. Biochim Biophys Acta Mol Cell Res 2018; 1865: 721-33.
28. Zhang H, Qi S, Song Y, Ling C. Artemisinin attenuates early renal damage on diabetic nephropathy rats through suppressing TGF-β1 regulator and activating the Nrf2 signaling pathway. Life Sci 2020; 256: 117966.
29. Conti I, Varano G, Simioni C, et al. miRNAs as influencers of cell-cell communication in tumor microenvironment. Cells 2020; 9: 220.
30. Shen Y, Tong ZW, Zhou Y, et al. Inhibition of lncRNA-PAX8-AS1-N directly associated with VEGF/TGF-β1/8-OhdG enhances podocyte apoptosis in diabetic nephropathy. Eur Rev Med Pharmacol Sci 2020; 24: 6864-72.
31. Dong H, Qiang Z, Chai D, et al. Nrf2 inhibits ferroptosis and protects against acute lung injury due to intestinal ischemia reperfusion via regulating SLC7A11 and HO-1. Aging (Albany NY) 2020; 12: 12943-59.
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