丹参注射液调控TGF-β/Smad信号通路抑制腹膜透析液诱导HMrSV5细胞内皮-间充质转化机制

doi:10.12122/j.issn.1673-4254.2024.12.02

南方医科大学学报 ›› 2024, Vol. 44 ›› Issue (12): 2276-2282.doi: 10.12122/j.issn.1673-4254.2024.12.02

• • 上一篇

丹参注射液调控TGF-β/Smad信号通路抑制腹膜透析液诱导HMrSV5细胞内皮-间充质转化机制

俞丽华¹(), 李静雅¹(), 王晓琪², 李莉¹, 陈娅³, 王飞宇², 张坤²(), 王桐生¹()

^1.安徽中医药大学，中西结合学院生理与药理学系，安徽合肥 230012
^2.安徽中医药大学，药学院，安徽合肥 230012
^2.安徽中医药大学附属太和中医院肾病科，安徽太和 236600

收稿日期:2024-07-26 出版日期:2024-12-20 发布日期:2024-12-26
通讯作者: 张坤,王桐生 E-mail:lihua2831699@163.com;ljy20210202@163.com;qixf2008@163.com;wtsyl@163.com
作者简介:俞丽华，主治医师，E-mail: lihua2831699@163.com
李静雅，在读博士研究生，E-mail: ljy20210202@163.com
第一联系人：俞丽华、李静雅共同第一作者
基金资助:
国家自然科学基金(82174162);安徽省高校自然科学研究项目(2023AH050847)

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

Lihua YU¹(), Jingya LI¹(), Xiaoqi WANG², Li LI¹, Ya CHEN³, Feiyu WANG², Kun ZHANG²(), Tongsheng WANG¹()

^1.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
^2.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
^3.Department of Nephrology, Taihe Traditional Chinese Medicine Hospital Affiliated to Anhui University of Chinese Medicine, Taihe 236600, China

Received:2024-07-26 Online:2024-12-20 Published:2024-12-26
Contact: Kun ZHANG, Tongsheng WANG E-mail:lihua2831699@163.com;ljy20210202@163.com;qixf2008@163.com;wtsyl@163.com

摘要/Abstract

摘要：

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。

关键词: 丹参注射液, 腹膜透析液, 内皮-间充质转化, TGF-β/Smad 信号通路, 腹膜纤维化

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

俞丽华, 李静雅, 王晓琪, 李莉, 陈娅, 王飞宇, 张坤, 王桐生. 丹参注射液调控TGF-β/Smad信号通路抑制腹膜透析液诱导HMrSV5细胞内皮-间充质转化机制[J]. 南方医科大学学报, 2024, 44(12): 2276-2282.

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.

图/表 5

图1 腹膜透析液对HMrSV5细胞活力影响

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

图2 腹膜透析液对HMrSV5细胞E-cadherin、α-SMA蛋白表达的影响

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.

图3 丹参注射液对HMrSV5细胞活力影响

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

图4 丹参注射液对腹膜透析液诱导HMrSV5细胞上清液中IL-6、TNF-α、TGF-β、VEGF水平影响

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.

图5 丹参注射液对腹膜透析液诱导HMrSV5细胞E-cadherin、α-SMA、p-Smad 2/3、Smad 7蛋白表达影响

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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丹参注射液调控TGF-β/Smad信号通路抑制腹膜透析液诱导HMrSV5细胞内皮-间充质转化机制

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

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