云锡矿粉诱导成纤维细胞活化蛋白表达及与上皮细胞共培养对其表达的影响  

A study on relationship between expressions of FAP protein in fibroblasts induced by Yunnan tin mine dust and co-cultivation of epithelia

作　　者：边莉[1] 何永文[2] 阮永华[3] 高倩[3] 王春艳[1] 金克炜[3] 

机构地区：[1]昆明医学院第一附属医院病理科,云南昆明650032 [2]昆明医学院口腔医学研究所,650031 [3]昆明医学院基础医学院,650031

出　　处：《中国职业医学》2010年第1期1-3,7,共4页China Occupational Medicine

基　　金：国家自然科学基金项目(30560053);云南省应用基础研究项目(2008CD009;2006C0042M);教育部高等学校博士学科点专项科研基金项目(20050678004)

摘　　要：目的研究云锡矿粉诱导成纤维细胞活化蛋白(FAP)表达及与上皮细胞共培养对其表达的影响。方法选用100 mg/L的云锡矿粉处理永生化人支气管上皮细胞(BEAS-2B)及人胚肺成纤维细胞(WI-38),每次处理72 h,隔代处理直至第9代。自第11代将上述细胞分组共培养。分组如下:W组、W(B)组、W(B100)组、W100组、W100(B)组、W100(B100)组。应用免疫印迹检测第16代、26代及36代各组成纤维细胞中FAP的表达,以β-actin作为内参,结果通过Quantity One软件进行分析,FAP水平=样本FAP灰度值/同一样本β-actin灰度值。结果免疫印迹检测WI-38中无FAP表达;第16代各组WI-38均未检测到FAP表达;至第26代时,W(B)组及W(B100)组FAP仍呈阴性表达,而W100组、W100(B)组及W100(B100)组则可检测到FAP表达,但3组间表达水平差异无统计学意义(P>0.05)。第36代时,W100组、W100(B)组及W100(B100)组FAP表达水平较第26代增强,但W100组和W100(B)组FAP表达水平无明显差异,惟W100(B100)组FAP表达水平明显强于前2组(P<0.05);且W(B100)组也出现了FAP表达。结论云锡矿粉能诱导WI-38表达FAP,矿粉诱导的上皮细胞可提高其表达水平。Objective To study the expressions of fibroblast activation protein（FAP） in fibroblasts induced by Yunnan tin mine dust and co-cultivation of epithelia.Methods Every other generation of immortalized human bronchial epithelial cells（BEAS-2B） and human embryo lung fibroblasts（WI-38） were treated with Yunnan tin mine dust at 100 mg/L for 72 hours,until the ninth generation.Then from the 11th generation,the cells were subsequently co-cultured as follows：W group、W（B） group、W（B100） group、W100 group、W100（B） group;W100（B100） group.The expressions of FAP in fibroblasts were determined by the method of immunoblot at the 16th,26th and 36th generation.The expressions of β-actin were used as the control.The results were analyzed with Quantity One software.The quantity of FAP protein =gray value of FAP/gray value of β-actin.Results The expression of FAP was not observed in any generation of normal fibroblasts.or in any of the groups At the 16th generation,nor in the W（B） and W（B100）groups At the 26th generation,but positive expressions were observed in the W100,W100（B） and W100（B100） groups,although no statistical significance were shown among groups.At the 36th generation,the expressions of FAP in W100,W100（B）and W100（B100）groups were increased compared with those at the 26th generation.There was no statistical significance between the expression levels of W100 and W100（B）groups,but the expression level of W100（B100） group was much higher than those of W100 and W100（B）groups（P0.05）.Furthermore,the expression of FAP was also observed in W（B100） group.Conclusion Tin mine dust can induce expressions of FAP in fibroblasts and the expression level of FAP can be promoted by epithelia induced by Yunnan tin mine dust.

关 键 词：云锡矿粉 人胚肺成纤维细胞(WI-38) 成纤维细胞活化蛋白(FAP) 人支气管上皮细胞(BEAS-2B) 

分 类 号：R135.2[医药卫生—劳动卫生]