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Baicalin suppresses renal fibrosis through microRNA-124/TLR4/NF-κB axis in streptozotocin-induced diabetic nephropathy mice and high glucose-treated human proximal tubule epithelial cells

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Abstract

Renal fibrosis is a major pathological event in the development of diabetic nephropathy (DN). Baicalin is a flavonoid glycoside that possesses multiple pharmacological properties including anti-fibrotic activity. In the present study, the effects of baicalin on renal fibrosis along with related molecular basis were investigated in streptozotocin (STZ)-induced DN mouse model and high glucose (HG)-treated HK-2 human proximal tubule epithelial cell model. Renal injury was evaluated through blood urea nitrogen (BUN) and serum creatinine (Scr) levels and urine albumin creatine ratio (ACR). Renal fibrosis was assessed by type IV collagen (COLIV) and fibronectin (FN) protein expression and histopathologic analysis via Masson trichrome staining. Protein levels of COLIV, FN, NF-κB inhibitor alpha (IκBα), phosphorylated IκBα (p-IκBα), p65, phosphorylated p65 (p-p65), and toll-like receptor 4 (TLR4) were measured by western blot assay. MicroRNA-124 (miR-124) and TLR4 mRNA levels were detected by RT-qPCR assay. The interaction of miR-124 and TLR4 was examined by bioinformatics analysis, luciferase reporter assay, and RIP assay. Baicalin or miR-124 attenuated renal injury and fibrosis in STZ-induced DN mice. Baicalin inhibited the increase of COLIV and FN expression induced by HG through upregulating miR-124 in HK-2 cells. TLR4 was a target of miR-124. MiR-124 inhibited TLR4/NF-κB pathway activation and the inactivation of the NF-κB pathway hindered COLIV and FN expression in HG-stimulated HK-2 cells. Baicalin prevented renal fibrosis by increasing miR-124 and inactivating downstream TLR4/NF-κB pathway in DN, hinting the pivotal values of baicalin and miR-124 in the management of DN and renal fibrosis.

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Funding

This research was supported by the Henan Research Institute of Chinese Medicine Pre-research Fund (No. 1704570).

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Authors and Affiliations

  1. Henan Academy of Chinese Medicine, Zhengzhou, 450000, China

    Shefeng Zhang, Ruifeng Liang & Chenhua Yang

  2. Henan University of Chinese Medicine, Zhengzhou, 450000, China

    Li Xu

  3. The First Affiliated Hospital of Henan University of Chinese Medicine, No. 19, Renmin Road, Jinshui District, Zhengzhou, 450000, China

    Peiren Wang

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  1. Shefeng Zhang
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  2. Li Xu
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  3. Ruifeng Liang
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  4. Chenhua Yang
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Contributions

Shefeng Zhang designed and performed the experiments, wrote the manuscript. Li Xu and Ruifeng Liang contributed to experimental work and data analysis. Chenhua Yang and Peiren Wang conducted the experiments and revised the manuscript. All authors have read and approved the final manuscript.

Corresponding author

Correspondence to Peiren Wang.

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All animal experiments were carried out with the approval of Animal Ethics Committee of The First Affiliated Hospital of Henan University of Chinese Medicine and the standard for the Care and Use of Laboratory Animals by the National Institutes of Health Guide.

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The authors declare that they have no conflict of interest.

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Key points

• Baicalin or miR-124 overexpression attenuated renal fibrosis in STZ-induced DN mice.

• Baicalin inhibited the expression of fibrosis markers by upregulating miR-124 in HG-exposed HK-2 cells.

• MiR-124 inhibited the activation of the NF-κB pathway by targeting TLR4.

• Baicalin prevented renal fibrosis by regulating the miR-124/TLR4/NF-κB pathway in DN.

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Zhang, S., Xu, L., Liang, R. et al. Baicalin suppresses renal fibrosis through microRNA-124/TLR4/NF-κB axis in streptozotocin-induced diabetic nephropathy mice and high glucose-treated human proximal tubule epithelial cells. J Physiol Biochem 76, 407–416 (2020). https://doi.org/10.1007/s13105-020-00747-z

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  • DOI: https://doi.org/10.1007/s13105-020-00747-z

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