Study on the Effect and Mechanism of Danggui Buxue Decoction Inhibiting HK-2 Cell Fibrosis through miR-27a/TGF-β1/Smad3 Pathway

  OBJECTIVE  To investigate the mechanism of Danggui Buxue Decoction regulating HK-2 cell fibrosis through miR-27a.

  METHODS  HK-2 cells were divided into control group, high glucose group, high glucose+blank serum group, and high glucose+Danggui Buxue Decoction-containing serum group. The proliferation of cells in each group was detected by MTT assay; The expression of the fibrosis-related factors Vimentin, COL Ⅳ, α-SMA, p-Smad3/Smad3 and TGF-β1 protein was measured by Western blot; The qPCR was used to detect the expression of miR-27a and fibrosis-related factors mRNA; The expression of miR-27a and fibrosis-related proteins were observed by transfecting miR-27a inhibitor and Smad3-siRNA, as well as adding TGF-β1 to the culture medium.

  RESULTS  MTT results showed that compared with the control group, the cell proliferation rate in the high glucose group significantly decreased (P < 0.05, P < 0.001), and Danggui Buxue Decoction significantly promoted cell proliferation (P < 0.05, P < 0.001). Western blot showed that Danggui Buxue Decoction significantly inhibited the expression of fibrosis-related proteins COL Ⅳ, α-SMA, Vimentin, TGF-β1 and p-Smad3/Smad3 in HK-2 cells induced by high glucose (P < 0.05, P < 0.01). The qPCR results were consistent with Western blot results. After TGF-β1 treatment of cells, cell fibrosis aggravated, and the expression of miR-27a increased (P < 0.05, P < 0.01), Danggui Buxue Decoction significantly reduced the elevation of miR-27a in TGF-β1-induced cellular fibrosis (P < 0.001). Transfection of miR-27a inhibitor and Smad3-siRNA could significantly reduce the increase of TGF-β1-induced cellular fibrosis protein expression (P < 0.05, P < 0.01).

  CONCLUSION  Danggui Buxue Decoction-containing serum can promote the proliferation of HK-2 cells and regulate HK-2 cell fibrosis through miR-27a/TGF-β1/Smad3 signaling pathway.