Open AccessTanshinone IIA improves diabetes‐induced renal fibrosis by regulating the miR ‐34‐5p/Notch1 axis
Author(s) -
Zhang Lizhi,
Yang Fan
Publication year - 2022
Publication title -
food science and nutrition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.614
H-Index - 27
ISSN - 2048-7177
DOI - 10.1002/fsn3.2998
Subject(s) - apoptosis , in vivo , western blot , tunel assay , chemistry , flow cytometry , microbiology and biotechnology , fibrosis , cell , microrna , cell growth , reporter gene , in vitro , pathology , biology , gene expression , medicine , biochemistry , gene
The purpose of this study was to evaluate the improvement of tanshinone in renal fibrosis in vitro and in vivo study. It used streptozotocin to model diabetic nephropathy (DN) mice, and treated with different Tanshinone IIA concentrations. The pathology of kidney tissues was evaluated by hematoxylin and eosin (H&E) and Masson's staining; the ultrastructure and apoptosis cell number of kidney tissues were evaluated by transmission electron microscopy (TEM) and TUNEL assay. Relative gene and protein expression was evaluated by reverse transcription‐quantitative polymerase chain reaction (RT‐qPCR), immunohistochemical (IHC) analysis, or western blot (WB) assay. In vitro study, using high‐glucose stimulated HK‐2 cell to model DN cell model, measuring cell proliferation, apoptosis rate, relative gene and protein expression, and LC 3B and P62 proteins expression by Cell Counting Kit‐8 (CCK‐8), flow cytometry, RT‐qPCR, WB, and cell immunofluorescence. Analysis correlation between Notch1 and miRNA‐34a‐5p was carried out by dual‐luciferase reporter. Fibrosis area and apoptosis cell rate were significantly up‐regulated ( p < .001), with Tanshinone IIA supplement. The fibrosis area and apoptosis cell rate were also significantly improved in a dose‐dependent manner ( p < .05). With si‐miRNA‐34a‐5p transfection, the Tanshinone IIA's treatment effects were significantly depressed. By dual‐luciferase reporter, miRNA‐34a‐5p could target Notch1 in the HK‐2 cell line. Tanshinone IIA improved DN‐induced renal fibrosis by regulating miRNA‐34a‐5p in vitro and in vivo study.