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-p/Smad signaling pathway

作　　者：俞丽华[1] 李静雅[1] 王晓琪 李莉[1] 陈娅 王飞宇 张坤 王桐生[1] 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,School of Pharmacy,Anhui University of Chinese Medicine,Hefei 230012,China;Department of Nephrology,Taihe Traditional Chinese Medicine Hospital Affiliated to Anhui University of Chinese Medicine,Taihe 236600,China)

机构地区：[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 [3]安徽中医药大学附属太和中医院肾病科,安徽太和236600

出　　处：《南方医科大学学报》2024年第12期2276-2282,共7页Journal of Southern Medical University

基　　金：国家自然科学基金(82174162);安徽省高校自然科学研究项目(2023AH050847)。

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、Westernblotting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72h诱发细胞EndMT,实验结束前24h,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;Westernblotting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-Smad2/3和Smad7蛋白表达影响。结果40%的腹膜透析液诱导HMrSV5细胞72h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。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-βand VEGF,upregulated the protein expressions ofα-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-βand VEGF and the protein expressions of aα-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.

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 

分 类 号：R285[医药卫生—中药学]