Yu Lihua, Li Jingya, Wang Xiaoqi, Li Li, Chen Ya, Wang Feiyu, Zhang Kun, Wang Tongsheng
Department of Physiology and Pharmacology, School of Integrated Chinese and Western Medicine, Taihe Traditional Chinese Medicine Hospital Affiliated to Anhui University of Chinese Medicine, Taihe 236600, China.
School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, China, Taihe Traditional Chinese Medicine Hospital Affiliated to Anhui University of Chinese Medicine, Taihe 236600, China.
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.
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.
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.
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.
Danshen Injection can suppress peritoneal dialysis fluid-induced EndMT in HMrSV5 cells possibly by regulating the TGF-β/Smad signaling pathway.
研究丹参注射液对腹膜透析液诱导的HMrSV5细胞内皮-间充质转化(EndMT)的抑制作用以及转化生长因子-β(TGF-β)/Smad信号通路在介导该作用中的作用。
将培养于40%腹膜透析液中72小时以诱导EndMT的HMrSV5细胞分别用0.05%、0.1%和0.5%的丹参注射液处理。采用CCK-8法评估处理后细胞活力的变化,用酶联免疫吸附测定法(ELISA)检测细胞上清液中白细胞介素-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)的水平;进行蛋白质免疫印迹法检测细胞中E-钙黏蛋白、α-平滑肌肌动蛋白(α-SMA)、磷酸化Smad 2/3和Smad 7的蛋白表达。
在40%腹膜透析液中培养72小时可诱导HMrSV5细胞发生明显的EndMT,细胞活力明显降低。浓度范围在0.025%-1.5%的丹参注射液对细胞活力无显著影响。HMrSV5细胞暴露于腹膜透析液72小时后,IL-6、TNF-α、TGF-β和VEGF的产生显著增加,α-SMA和磷酸化Smad 2/3的蛋白表达上调,E-钙黏蛋白和Smad7蛋白表达降低。用丹参注射液处理暴露的细胞可显著提高细胞活力,增加E-钙黏蛋白和Smad 7蛋白的细胞表达,并减少IL-6、TNF-α、TGF-β和VEGF的产生以及α-SMA和磷酸化Smad 2/3的蛋白表达。
丹参注射液可能通过调节TGF-β/Smad信号通路抑制腹膜透析液诱导的HMrSV5细胞EndMT。