人胎盘间充质干细胞调控TGF-β1/Smad3信号通路抑制肺纤维化的发生  

Human placental mesenchymal stem cells inhibit occurrence of pulmonary fibrosis by regulating transforming growth factor-beta 1/Smad3 signaling pathway

作　　者：曹佳伟 丁劭瑞 铁华 薛菁[2,3] 贾元元[2,3] 梁雪云[3] 李锋 Cao Jiawei;Ding Shaorui;Tie Hua;Xue Jing;Jia Yuanyuan;Liang Xueyun;Li Feng(Clinical Medical College,Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;General Hospital of Ningxia Medical University,Ningxia Academy of Medical Sciences,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Key Laboratory of Stem Cell and Regenerative Medicine of Ningxia Hui Autonomous Region,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Medical Experimental Center,General Hospital of Ningxia Medical University,Yinchuan 750004,Ningxia Hui Autonomous Region,China)

机构地区：[1]宁夏医科大学临床医学院,宁夏回族自治区银川市750004 [2]宁夏医学科学研究院,宁夏医科大学总医院,宁夏回族自治区银川市750004 [3]宁夏回族自治区干细胞与再生医学重点实验室,宁夏医科大学总医院,宁夏回族自治区银川市750004 [4]宁夏医科大学总医院医学实验中心,宁夏回族自治区银川市750004

出　　处：《中国组织工程研究》2024年第31期4970-4974,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金项目(81860566),项目负责人:李锋。

摘　　要：背景:研究表明人胎盘间充质干细胞能有效抑制肺纤维化的发展,但其发挥作用的具体机制目前并不清楚。目的:探究人胎盘间充质干细胞对二氧化硅诱导人胚肺成纤维细胞发生肺纤维化的治疗作用及相关机制。方法:CCK-8法检测不同质量浓度二氧化硅干预不同时间对MRC-5细胞增殖活力的影响,结合免疫荧光染色法筛选出最佳的二氧化硅刺激质量浓度与时间用于后续实验。将MRC-5细胞分为空白组、二氧化硅组、二氧化硅+人胎盘间充质干细胞组,空白组细胞不予任何处理,二氧化硅组MRC-5细胞给予100μg/mL二氧化硅刺激48 h,二氧化硅+人胎盘间充质干细胞组MRC-5细胞先给予100μg/mL二氧化硅刺激48 h后再与人胎盘间充质干细胞共培养24 h。免疫荧光染色检测各组细胞α-平滑肌肌动蛋白、胶原蛋白Ⅰ型蛋白的表达;Western blot检测各组细胞肺纤维化相关蛋白和TGF-β1/Smad 3信号通路相关蛋白表达。结果与结论:①CCK-8结果显示,100μg/mL二氧化硅刺激MRC-5细胞48 h为最佳质量浓度与时间;②免疫荧光染色结果显示,与二氧化硅组相比,二氧化硅+人胎盘间充质干细胞组α-平滑肌肌动蛋白、胶原蛋白Ⅰ型蛋白表达明显降低;③Western blot结果显示,与二氧化硅组相比,二氧化硅+人胎盘间充质干细胞组α-平滑肌肌动蛋白、胶原蛋白Ⅰ型、N-钙黏蛋白、粘连蛋白、转化生长因子β1、p-Smad3、Smad3的蛋白表达均降低,E-钙黏蛋白表达升高,差异均有显著性意义(P<0.05);④结果说明,人胎盘间充质干细胞能够对二氧化硅诱导的肺纤维化有显著的治疗作用;人胎盘间充质干细胞可以通过调控TGF-β1/Smad3信号通路抑制肺纤维化的发生。BACKGROUND:Human placental mesenchymal stem cells have been shown to be effective in inhibiting the development of pulmonary fibrosis,but the underlying mechanisms remain unclear.OBJECTIVE:To investigate the therapeutic effect and related mechanism of human placental mesenchymal stem cells on silica-induced pulmonary fibrosis in human embryonic lung fibroblasts(MRC-5).METHODS:CCK-8 assay was used to detect the effects of different mass concentrations of silica on the proliferation of MRC-5 at different time points.Immunofluorescence staining was used to screen out the best stimulating mass concentration and time of silica for subsequent experiments.MRC-5 cells were divided into blank group,silica group,and silica+human placental mesenchymal stem cell group.In the blank group,cells were not treated.In the silica group,MRC-5 cells were stimulated with 100μg/mL silica for 48 hours.In the silica+human placental mesenchymal stem cell group,MRC-5 cells were stimulated with 100μg/mL silica for 48 hours and then co-cultured with human placental mesenchymal stem cells for 24 hours.Immunofluorescence staining was used to detect the expression ofα-smooth muscle actin and collagen type I in cells of each group.Western blot assay was used to detect the expressions of pulmonary fibrosis-related proteins and TGF-β1/Smad 3 signaling pathway-related proteins in cells of each group.RESULTS AND CONCLUSION:(1)CCK-8 assay results suggested that 100μg/mL silica was the best mass concentration and time to stimulate MRC-5 cells for 48 hours.(2)Immunofluorescence staining results showed that the expression ofα-smooth muscle actin and collagen type I in the silica+human placental mesenchymal stem cell group was significantly lower than that in the silica group.(3)Western blot assay results showed that compared with the silica group,the protein expression levels ofα-smooth muscle actin,collagen type I,N-cadherin,fibronectin,transforming growth factor-β1,p-Smad3,and Smad3 in the silica+human placental mesenchymal stem cell group wer

关 键 词：人胎盘间充质干细胞 肺纤维化 MRC-5细胞 转化生长因子Β1 SMAD3 信号通路 

分 类 号：R456[医药卫生—治疗学] R34[医药卫生—临床医学] R11