PD98059抑制哮喘中TGF-β1诱导的人支气管上皮细胞上皮间质转化进程  

PD98059 inhibits TGF-β1-induced epithelial-mesenchymal transition of human bronchial epithelial cells in asthma

作　　者：吴文棋 庄旭辉 王姝晨 李静静 王霞[1,3] 王勇[3] 马武华[3] WU Wenqi;ZHUANG Xuhui;WANG Shuchen;LI Jingjing;WANG Xia;WANG Yong;MA Wuhua(Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006;China.2.Department of Anesthesiology,Sun Yat-sen Memorial Hospital,Sun Yat-sen University,Guangzhou 510000;.3.Department of Anesthesiology,First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine,Guangzhou 510006;.4.Jincheng Department of Anesthesiology,Municipal People’s Hospital,Jincheng 048000)

机构地区：[1]广州中医药大学,广州510006 [2]中山大学孙逸仙纪念医院麻醉科,广州510000 [3]广州中医药大学第一附属医院麻醉科,广州510006 [4]晋城市人民医院麻醉科,山西晋城048000

出　　处：《中国比较医学杂志》2022年第7期34-40,共7页Chinese Journal of Comparative Medicine

基　　金：国家自然科学基金(81673922,82074357)。

摘　　要：目的探究PD98059对转化生长因子β1(TGF-β1)诱导人支气管上皮细胞BEAS-2B上皮间质转化(EMT)进程的影响及可能的分子机制。方法本研究采用人支气管上皮细胞(BEAS-2B),使用TGF-β1诱导BEAS-2B上皮间质转化模型,使用ERK1/2抑制剂PD98059探究抑制ERK/MAPK信号转导是否影响EMT进程。通过免疫荧光检测BEAS-2B气道重塑模型α-SMA蛋白的表达及定位;CCK-8法检测各组细胞存活率;Edu实验检测各组细胞增殖能力水平;transwell实验检测各组细胞迁移能力水平;Western blot检测各组EMT标志蛋白表达及ERK/MAPK通路的激活情况。结果与正常组相比,模型组α-SMA表达增加,α-SMA在BEAS-2B细胞胞质上表达。PD98059在40μmol/L浓度与5 ng/mL TGF-β1共处理抑制BEAS-2B细胞生长(P<0.05)。与三组Edu阳性率无显著差异(P>0.05)。与正常组相比,模型组迁移细胞数增加(P<0.05),上皮间质化标志蛋白表达增加(P<0.05),p-ERK蛋白表达增加(P<0.05),PD98059可以明显减少迁移细胞数(P<0.05),抑制上皮间质化标志蛋白的表达(P<0.05)及ERK通路的激活(P<0.05)。结论TGF-β1可能通过ERK/MAPK通路促进人支气管上皮细胞上皮间质转化进程,PD98059可以通过抑制ERK/MAPK通路的激活抑制支气管上皮细胞EMT进程。Objective To investigate the effect of PD98059 on epithelial-mesenchymal transition(EMT) of transforming growth factor β1(TGF-β1)-induced human bronchial epithelial cells BEAS-2 B and the possible molecular mechanism. Methods Human bronchial epithelial cells(BEAS-2 B) were used for the BEAS-2 B EMT model established with TGF-β1, and the ERK1/2 inhibitor PD98059 was used to explore whether inhibition of ERK/MAPK signal transduction affects the EMT process. Immunofluorescence was used to detect the expression and location of α-SMA protein in the BEAS-2 B airway remodeling model. CCK-8 assays were used to assess the cell survival rate. An Edu proliferation assay was used to assess cell proliferation. Transwell assays were used to assess cell migration. Western blott was used to detect the expression of EMT marker proteins and activation of the ERK/MAPK pathway. Results Compared with the control group, α-SMA expression in the model group was increased, and α-SMA was expressed in the cytoplasm of BEAS-2 B cells. Cotreatment with 40 μmol/L PD98059 and 5 ng/mL TGF-β1 inhibited BEAS-2 B cell growth(P<0.05). There was no significant difference in Edu-positive cells among the three groups(P>0.05). Compared with the normal group, the number of migrating cells(P<0.05), expression of EMT marker proteins(P<0.05), and expression of p-ERK protein were increased in the model group(P<0.05). PD98059 significantly reduced the number of migrating cells(P<0.05) and inhibited the expression of EMT marker proteins(P<0.05) and activation of the ERK pathway(P<0.05). Conclusions TGF-β1 promotes the EMT of human bronchial epithelial cells via the ERK/MAPK pathway. PD98059 inhibits EMT of bronchial epithelial cells by suppressing activation of the ERK/MAPK pathway.

关 键 词：哮喘 PD98059 ERK/MAPK 上皮间质转化 气道重塑 

分 类 号：R-33[医药卫生]