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

Nan Fang Yi Ke Da Xue Xue Bao. 2024 Dec 20;44(12):2276-2282. doi: 10.12122/j.issn.1673-4254.2024.12.02.
[Article in Chinese]

Abstract

Objectives: 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.

Conclusions: Danshen Injection can suppress peritoneal dialysis fluid-induced EndMT in HMrSV5 cells possibly by regulating the TGF-β/Smad signaling pathway.

目的: 研究丹参注射液对腹膜透析液诱导HMrSV5细胞发生内皮-间充质转化(EndMT)的影响,并从TGF-β/Smad信号通路分析其可能的作用机制。方法: CCK-8法、Western blotting筛选腹膜透析液诱导HMrSV5细胞发生EndMT的形成条件及丹参注射液安全浓度范围;取HMrSV5细胞,随机分成正常组、模型组、丹参注射液0.05%、0.1%、0.5%浓度组,除正常组外,其余各组加入40%腹膜透析液培养72 h诱发细胞EndMT,实验结束前24 h,各组加入不同浓度丹参注射液。CCK-8法检测细胞活力;ELISA法检测细胞上清中白细胞介素-6 (IL-6)、肿瘤坏死因子-α(TNF-α)、转化生长因子-β (TGF-β)和血管内皮生长因子(VEGF)水平;Western blotting检测各组细胞E-钙粘着蛋白(E-cadherin)、α-平滑肌激动蛋白(α-SMA)、p-Smad 2/3和Smad 7蛋白表达影响。结果: 40%的腹膜透析液诱导HMrSV5细胞72 h后,可发生明显的EndMT;丹参注射液安全浓度范围为0.025%~1.5%。与正常组比较,模型组细胞活力下降;IL-6、TNF-α、TGF-β和VEGF水平升高;α-SMA、p-Smad 2/3等蛋白表达上调;E-cadherin和Smad7蛋白表达明显下降,差异有统计学意义(P<0.05)。与模型组比,丹参注射液各浓度组可增加细胞活力,提高E-cadherin和Smad7蛋白表达,降低IL-6、TNF-α、TGF-β、VEGF水平和 α-SMA、p-Smad2/3蛋白表达(P<0.05)。结论: 丹参注射液对腹膜透析液诱导的EndMT有一定的改善作用,其机制可能与抑制TGF-β/Smad信号通路有关。.

Keywords: Danshen Injection; TGF‑β/Smad signaling pathway; endothelial-mesenchymal transition; peritoneal dialysate; peritoneal fibrosis.

Publication types

  • English Abstract

MeSH terms

  • Actins / metabolism
  • Cadherins / metabolism
  • Cell Line
  • Dialysis Solutions
  • Drugs, Chinese Herbal* / pharmacology
  • Endothelial-Mesenchymal Transition
  • Epithelial-Mesenchymal Transition* / drug effects
  • Humans
  • Interleukin-6 / metabolism
  • Peritoneal Dialysis* / adverse effects
  • Salvia miltiorrhiza*
  • Signal Transduction* / drug effects
  • Smad Proteins* / metabolism
  • Transforming Growth Factor beta* / metabolism
  • Tumor Necrosis Factor-alpha / metabolism
  • Vascular Endothelial Growth Factor A* / metabolism

Substances

  • Drugs, Chinese Herbal
  • Transforming Growth Factor beta
  • Smad Proteins
  • Vascular Endothelial Growth Factor A
  • Tumor Necrosis Factor-alpha
  • Interleukin-6
  • dan-shen root extract
  • Cadherins
  • Actins
  • Dialysis Solutions