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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 effect of tin-tin ore powder on the expression of fibroblast activating protein (FAP) and its co-culture with epithelial cells. Methods Immortalized human bronchial epithelial cells (BEAS-2B) and human embryonic lung fibroblasts (WI-38) were treated with 100 mg / L tin-yunksite powder and treated for 72 h each. The above cells were co-cultured from the 11th generation. Groups were as follows: W group, W (B) group, W (B100) group, W100 group, W100 (B) group, W100 (B100) group. The expression of FAP in each group of fibroblasts at the 16th, the 26th and the 36th generation was detected by immunoblotting. Β-actin was used as an internal control. The results were analyzed by Quantity One software. The FAP level was the same as the sample FAP gray value / the same sample β- actin gray value. Results The expression of FAP was not detected in WI-38 by Western blotting. FAP expression was not detected in WI-38 of the 16th generation. However, the expression of FAP was still negative in W (B) and W (B100) However, FAP expression was detected in W100 group, W100 (B) group and W100 (B100) group, but there was no significant difference among the three groups (P> 0.05). At the 36th generation, the expression of FAP in W100 group, W100 (B) group and W100 (B100) group was higher than that in the 26th generation, but there was no significant difference in FAP expression between W100 group and W100 (B) group FAP expression was significantly higher than the previous two groups (P <0.05); and W (B100) group also appeared FAP expression. Conclusion Cloud powder can induce the expression of FAP in WI-38, and its expression can be enhanced by mineral powder-induced epithelial cells.