硫酸铍诱导人肺腺癌A549细胞纤维化中细胞因子的变化  

Changes of Cytokines in Fibrosis of Human Lung Adenocarcinoma A549 Cells Induced by BeSO_(4)

作　　者：戚发秋 王凯 陈小惠 刘红娅 赵枫 付有娟 关素珍 QI Faqiu;WANG Kai;CHEN Xiaohui;LIU Hongya;ZHAO Feng;FU Youjuan;GUAN Suzhen(School of Public Health and Management,Ningxia Medical University,Yinchuan 750004,China;Ningxia Key Laboratory of Environmental Factors and Chronic Disease Control,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院,银川750004 [2]宁夏环境因素与慢性病控制重点实验室,银川750004

出　　处：《宁夏医科大学学报》2021年第12期1211-1214,1220,共5页Journal of Ningxia Medical University

基　　金：宁夏自然科学基金项目(2019AAC03085);宁夏医科大学2021年自治区级大学生创新创业训练计划项目(S202110752018)。

摘　　要：目的构建硫酸铍(BeSO_(4))致人肺腺癌A549细胞纤维化模型并验证转化生长因子-β1(TGF-β1)及其他纤维化相关因子的变化。方法取对数生长期的A549细胞,采用CCK-8法检测染毒不同浓度(0、1、10、50、100、150μmol·L^(-1))、不同时间(24、48、72 h)对细胞存活率的影响,确定后续实验染毒浓度与染毒时间。于倒置显微镜下分别观察细胞形态,并采用蛋白免疫印迹(Western blot)检测α-平滑肌肌动蛋白(α-SMA)、TGF-β1、Ⅰ型胶原蛋白(COL-Ⅰ)和Ⅲ型胶原蛋白(COL-Ⅲ)相对表达水平。结果随着BeSO_(4)浓度增加,A549细胞活力呈下降趋势(P均<0.05),且呈时间和剂量依赖性,确定BeSO_(4)的染毒浓度为1、10、100μmol·L^(-1)(溶剂为完全培养基,设为低、中、高剂量组)为后续实验浓度,染毒时间为48 h。BeSO_(4)刺激A549细胞后,细胞形态发生改变,呈长纤维状。不同剂量BeSO_(4)处理A549细胞48 h后α-SMA、TGF-β1、COL-Ⅰ和COL-Ⅲ的蛋白相对表达量均呈升高趋势。与空白对照组相比,低剂量组α-SMA、TGF-β1、COL-Ⅰ和COL-Ⅲ的蛋白相对表达量分别增长了0.23、0.38、0.26和0.48倍,中剂量组α-SMA、TGF-β1、COL-Ⅰ和COL-Ⅲ的蛋白相对表达量分别增长了0.40、0.93、0.42和0.95倍,高剂量组α-SMA、TGF-β1、COL-Ⅰ和COL-Ⅲ的蛋白相对表达量分别增长了0.69、1.24、0.62和1.31倍(P均<0.05)。结论BeSO_(4)致A549细胞纤维化模型构建成功,可为后续实验提供基础。Objective To investigate the changes of transforming growth factor-β1(TGF-β1)and other fibrosisrelated factors in the fibrosis of human lung adenocarcinoma A549 cells induced by BeSO_(4).Methods A549 cells in logarithmic growth phase were collected to assess the effects of BeSO_(4)with different concentrations(0,1,10,50,100,150μmol·L^(-1))and different time points(24,48,72 h)on cell survival rate by CCK-8 to determine the exposure concentration and exposure time of the follow-up experiment.Cell morphology was observed by inverted microscop,and the protein expression levels ofα-SMA,TGF-β1,COL-Ⅰand COL-Ⅲwere detected by western blot.Results With the increase of BeSO_(4)concentration,the viability of A549 cells showed a downward trend(P<0.05),and it was time-dependent and dase-dependent.The exposure concentrations were 1,10 and 100μmol·L^(-1)of BeSO_(4),and exposure time is 48 h.The solvent was complete medium.After BeSO_(4)stimulation,A549 cells were morphologically changed and showed long fibrous shape.Different doses of BeSO_(4)treated A549 cells for 48 h,the relative expression ofα-SMA,TGF-β1,COL-Ⅰand COL-Ⅲall showed an increasing trend.Compared to the control group,the relative protein expression levels ofα-SMA,TGF-β1,COL-Ⅰand COL-Ⅲin the low-dose group were increased by 0.23,0.38,0.26 and 0.48 times,respectively(P<0.05).The relative protein expression levels ofα-SMA,TGF-β1,COL-Ⅰand COL-Ⅲin the medium-dose group were increased by 0.40,0.93,0.42 and 0.95 times,respectively(P<0.05).The relative protein expression levels ofα-SMA,TGF-β1,COL-Ⅰand COL-Ⅲin the high-dose group were increased by 0.69,1.24,0.62 and 1.31 times,respectively(P all<0.05).Conclusion The model of BeSO_(4)-induced fibrosis in A549 cells provides the basis for the subsequent experimental study.

关 键 词：硫酸铍 肺纤维化 胶原蛋白 Α-平滑肌肌动蛋白 转化生长因子-Β1 

分 类 号：R114[医药卫生—卫生毒理学] R563[医药卫生—公共卫生与预防医学]