青蒿提取物对镉暴露致人肺上皮细胞恶性转化及对Wnt、β-catenin表达的影响  

Effect of Artemisia annua extract on malignant transformation of human lung epithelial cells induced by cadmium exposure and expression of Wnt and β-catenin

作　　者：李峻峰 龙晓莉[2] 张琨鹏 LI Jun-feng;LONG Xiao-li;ZHANG Kun-peng(Department of Respiratory and Critical Care Medicine,Yunyang County People's Hospital,Chongqing,404599,China;Respiratory Department,Yongchuan Hospital Affiliated to Chongqing Medical University,Chongqing,402160,China)

机构地区：[1]重庆市云阳县人民医院呼吸与危重症医学科,重庆404599 [2]重庆医科大学附属永川医院呼吸科,重庆402160

出　　处：《职业与健康》2022年第21期2905-2911,共7页Occupation and Health

基　　金：重庆市永川区科委自然科学基金项目(Ycstc,2017nc5022)。

摘　　要：目的 研究青蒿提取物对镉暴露致人肺上皮细胞恶性转化及Wnt、β-catenin表达的影响。方法 设人肺上皮细胞A549组、镉暴露组(100μmol/L CdCl_(2))及青蒿提取物低、中、高剂量人肺上皮细胞A549组(100μmol/L CdCl_(2)+15、30、60μmol/L的青蒿提取物);各组每孔设6个平行样,培养72 h。细胞计数试剂盒-8测定细胞增殖水平,Transwell室、划痕试验测定细胞侵袭水平,流式细胞仪测定细胞凋亡水平,实时荧光逆转录法及蛋白印法测定细胞Wnt、β-catenin水平。结果 与人肺上皮细胞A549组比较,镉暴露组OD值(0.81±0.07 vs 0.92±0.05)、存活率[(80.62±6.69)%vs(89.63±9.57)%]、穿膜数[(198.54±82.65)vs(980.92±89.05)个]、迁移距离[(35.69±9.63)vs(105.69±8.05)μm]、Wnt mRNA(1.12±0.32 vs 3.28±0.79)、β-catenin mRNA(1.15±0.26 vs 3.56±0.23)、Wnt蛋白(0.26±0.05 vs 1.24±0.06)、β-catenin蛋白(0.34±0.07 vs 1.32±0.06)表达水平均升高(均P<0.05),凋亡率降低[(2.17±0.45)%vs(0.90±0.52)%](P<0.05);与镉暴露组比较,青蒿提取物各剂量组OD值、存活率、穿膜数、迁移距离、Wnt、β-catenin mRNA蛋白表达水平降低(均P<0.05),凋亡率升高[(0.90±0.52)%vs(1.22±0.39)%、(2.39±0.48)%、(4.12±0.51)%](P<0.05),且随着青蒿提取物干预剂量的增加,青蒿提取物各剂量组OD值、存活率、穿膜数、迁移距离、Wnt、β-catenin mRNA蛋白表达水平逐渐降低,凋亡率逐渐升高,具有剂量-效应关系(P<0.05)。结论 青蒿提取物能抑制镉暴露下人肺上皮细胞恶性转化,其机制可能与青蒿提取物抑制Wnt、β-catenint的表达进而抑制Wnt/β-catenin通路的激活有关。Objective To study the effect of Artemisia annua extract on the malignant transformation of human lung epithelial cells induced by cadmium exposure and the expression of Wnt andβ-catenin.Methods There were human lung epithelial cell A549group,cadmium exposure group(100μmol/L CdCl_(2)),low,medium and high dose of Artemisia annua extract human lung epithelial cell A549 group(100μmol/L CdCl_(2)+15,30,60μmol/L Artemisia annua extract),and parallel samples were set up for each well in each group,cultured for 72 h.The cell counting kit-8 was used to measure cell proliferation level.Transwell chamber and scratch test were used to measure cell invasion levels.The flow cytometry was used to measure cell apoptosis levels.The real-time fluorescence reverse transcription and western blotting were used to measure cell Wnt andβ-catenin levels.Results Compared with the human lung epithelial cell A549 group,the OD value(0.81±0.07 vs 0.92±0.05),survival rate[(80.62±6.69)%vs(89.63±9.57)%],transmembrane number[(198.54±82.65)vs(980.92±89.05)cells],migration distance[(35.69±9.63)vs(105.69±8.05)μm],and levels of Wnt mRNA(1.12±0.32 vs 3.28±0.79),β-catenin mRNA(1.15±0.26 vs 3.56±0.23),Wnt protein(0.26±0.05 vs1.24±0.06),β-catenin protein(0.34±0.07 vs 1.32±0.06)in the cadmium exposure group were increased(all P<0.05),and apoptosis rate were decreased[(2.17±0.45)%vs(0.90±0.52)%](P<0.05).Compared with the cadmium exposure group,the OD value,survival rate,transmembrane number,migration distance,Wnt,β-catenin mRNA protein expression levels in Artemisia annua extract groups of were decreased(all P<0.05),while the apoptosis rate were increased[(0.90±0.52)%vs(1.22±0.39)%,(2.39±0.48)%,(4.12±0.51)%](P<0.05).With the increase of the intervention dose of Artemisia annua,the OD value,survival rate,transmembrane number,migration distance,Wnt,β-catenin mRNA protein expression levels in each dose group of Artemisia annua extract gradually decreased,and the apoptosis rate gradually increased,with a dose-effect relationship

关 键 词：青蒿提取物 肺上皮细胞 镉暴露 恶性转化 WNT β-catenin 

分 类 号：R453[医药卫生—治疗学]