Danshen Injection inhibits peritoneal dialysis fluid-induced endothelial-mesenchymal transition in HMrSV5 cells by regulating the TGF-β/Smad signaling pathway

doi:10.12122/j.issn.1673-4254.2024.12.02

Abstract

Abstract:

Objective To investigate the inhibitory effect of Danshen Injection on endothelial-mesenchymal transition (EndMT) induced by peritoneal dialysis fluid in HMrSV5 cells and the role of the TGF‑β/Smad signaling pathway in mediating this effect. Methods HMrSV5 cells cultured in 40% peritoneal dialysis solution for 72 h to induce EndMT were treated with 0.05%, 0.1% and 0.5% Danshen Injection. CCK-8 assay was used to assess the changes in viability of the treated cells, and the levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), and vascular endothelial growth factor (VEGF) in the cell supernatant were detected using ELISA; Western blotting was performed to detect the protein expressions of E-cadherin, α-smooth muscle actin (α-SMA), p-Smad 2/3, and Smad 7 in the cells. Results Culture in 40% peritoneal dialysis fluid for 72 induced significant EndMT in HMrSV5 cells, which exhibited obviously lowered cell viability. Danshen Injection within the concentration range of 0.025%-1.5% did not significantly affect the viability of the cells. Exposure of HMrSV5 cells to peritoneal dialysis fluid for 72 h significantly increased the production of IL-6, TNF‑α, TGF‑β and VEGF, upregulated the protein expressions of α‑SMA and p-Smad 2/3, and lowered the expressions of E-cadherin and Smad7 proteins. Treatment of the exposed cells with Danshen injection significantly increased cell viability and cellular expressions of E-cadherin and Smad 7 proteins and reduced the production of IL-6, TNF-α, TGF-β and VEGF and the protein expressions of α‑SMA and p-Smad 2/3. Conclusion Danshen Injection can suppress peritoneal dialysis fluid-induced EndMT in HMrSV5 cells possibly by regulating the TGF-β/Smad signaling pathway.

Key words: Danshen Injection, peritoneal dialysate, endothelial-mesenchymal transition, TGF?β/Smad signaling pathway, peritoneal fibrosis

Lihua YU, Jingya LI, Xiaoqi WANG, Li LI, Ya CHEN, Feiyu WANG, Kun ZHANG, Tongsheng WANG. Danshen Injection inhibits peritoneal dialysis fluid-induced endothelial-mesenchymal transition in HMrSV5 cells by regulating the TGF-β/Smad signaling pathway[J]. Journal of Southern Medical University, 2024, 44(12): 2276-2282.

Figures/Tables 5

Fig.1 Effect of peritoneal dialysis fluid on HMrSV5 cell viability (n=3). *P<0.05, **P<0.01 vs Control group.

Fig.2 Effect of peritoneal dialysis solution (PDS) on protein expression levels of E-cadherin and α-SMA in HMrSV5 cells (n=3). *P<0.05, **P<0.01 vs Control group.

Fig.3 Effect of Danshen Injection on HMrSV5 cell viability (n=3). **P<0.01 vs Control group. ##P<0.01 vs Model group.

Fig.4 Effect of Danshen Injection on peritoneal dialysis fluid-induced elevation of supernatant IL-6, TNF-α, TGF-β and VEGF levels in HMrSV5 cells (n=3). *P<0.05, **P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group.

Fig.5 Effect of Danshen Injection on peritoneal dialysate-induced changes in protein expressions of E-cadherin, α-SMA, p-Smad 2/3 and Smad 7 in HMrSV5 cells (n=3). *P<0.05, **P<0.01 vs Control group; #P<0.05, ##P<0.01 vs Model group.

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