1,25-(OH)2D3 and its analogue BXL-628 inhibit high glucose-induced activation of RhoA/ROCK pathway in HK-2 cells

Zhang,Wei; Yi,Bin; Zhang,Ke; Li,Aimei; Yang,Shikun; Huang,Jing; Liu,Jishi; Zhang,Hao

doi:10.3892/etm.2017.4211

1,25-(OH)2D3 and its analogue BXL-628 inhibit high glucose-induced activation of RhoA/ROCK pathway in HK-2 cells

Authors:
- Wei Zhang
- Bin Yi
- Ke Zhang
- Aimei Li
- Shikun Yang
- Jing Huang
- Jishi Liu
- Hao Zhang
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Affiliations: Department of Nephrology, Xiangya Third Hospital of Central South University, Changsha, Hunan 410013, P.R. China
Published online on: March 9, 2017 https://doi.org/10.3892/etm.2017.4211
Pages: 1969-1976

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Abstract

It has previously been reported that 1,25-(OH)2D3 inhibits high glucose-induced epithelial-to-mesenchymal transition (EMT) in HK-2 cells. However, the mechanism of this renoprotective action remains unclear. Elocalcitol (BXL‑628), a vitamin D analog, has been suggested to be effective on the RhoA/Rho associated protein kinase (ROCK) pathway, which serves a crucial role in high glucose‑induced EMT. The aim of the present study was to investigate the effect of 1,25‑(OH)2D3 and its analogue BXL‑628 on high glucose‑induced activation of the RhoA/ROCK pathway in human renal proximal tubular cells. HK‑2 cells were co‑treated with high glucose and either 1,25‑(OH)2D3 or BXL‑628. The RhoA expression levels and ROCK activity of the membrane were assessed via western blot analysis or immunofluorescence. α-smooth muscle actin (α‑SMA) and epithelial (E)‑cadherin were detected using western blotting and reverse transcription-quantitative polymerase chain reaction (RT‑qPCR), whereas collagen I and fibronectin levels were measured by ELISA and RT‑qPCR. The results demonstrated that 1,25‑(OH)2D3 and BXL‑628 both significantly downregulated the expression of active RhoA and ROCK activity induced by high glucose (P<0.05). Furthermore, the expressions of α‑SMA, collagen I, and fibronectin were significantly downregulated at both protein and mRNA (P<0.05) levels, whereas the expression of E‑cadherin was significantly increased (P<0.05) by 1,25‑(OH)2D3 or BXL‑628 treatment. In conclusion, the vitamin D receptor agonist 1,25‑(OH)2D3 and its analogue BXL‑628 were both able to attenuate high glucose‑induced EMT and extracellular matrix accumulation of HK‑2 cells by suppressing the RhoA/ROCK signaling pathway in vitro.

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