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目的观察表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)对二氧化硅(Si O2)所致肺上皮细胞间质转化的影响,并探讨表皮生长因子受体(EGFR)信号通路在肺纤维化上皮间质转化中的作用。方法体外制备矽肺细胞模型,用EGFR-TKI干预染尘肺上皮细胞。实验分为正常对照组(Si O2粉尘浓度为0μg/ml)、模型组(Si O2粉尘浓度为200μg/ml)、低剂量干预组(A1组,EGFR-TKI浓度20μmol/L)、中剂量干预组(A2组,EGFR-TKI浓度40μmol/L)、高剂量干预组(A3组,EGFR-TKI浓度80μmol/L),干预48 h后,于倒置光学显微镜观察细胞形态学改变,并收集不同时段细胞,用RT-PCR检测E-cadherin、α-SMA、EGFR mRNA表达水平,细胞免疫荧光方法检测蛋白表达的变化。结果 EGFR-TKI干预,上皮细胞转化停滞,表现为纤维状伪足变短,呈类圆形;E-cadherin mRNA和蛋白表达无明显变化,模型组、A1组、A2组、A3组之间差异无统计学意义(P>0.05);α-SMA mRNA、EGFR mRNA和其蛋白表达逐渐下降,各组之间差异具有统计学意义(P<0.05)。结论 EGFR-TKI能抑制上皮间质转化,其机制可能与下调α-SMA表达和抑制EGFR活性有关。由此推测,抑制EGFR信号通路的活化可能为临床治疗肺纤维化提供新的思路和途径。
Objective To observe the effect of epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) on the interstitial transformation of lung epithelial cells induced by silicon dioxide (Si 2 O) and to explore the role of epidermal growth factor receptor (EGFR) signaling pathway in Role of pulmonary fibrosis in epithelial-mesenchymal transition. Methods Silicosis cells were prepared in vitro and the lung epithelial cells were exposed to EGFR-TKI. The experiment was divided into normal control group (Si O2 dust concentration 0μg / ml), model group (Si O2 dust concentration 200μg / ml), low dose intervention group (A1 group, EGFR-TKI concentration 20μmol / L) (Group A2, EGFR-TKI concentration 40μmol / L) and high dose intervention group (group A3, EGFR-TKI concentration 80μmol / L). After intervention for 48 hours, the morphological changes of cells were observed by inverted light microscope. The expression of E-cadherin, α-SMA and EGFR mRNA was detected by RT-PCR. The protein expression was detected by immunofluorescence staining. Results EGFR-TKI intervention showed that the transition of epithelial cells was stagnant. The appearance of fibrous pseudopods became short and round. The expression of E-cadherin mRNA and protein did not change significantly. The difference between model group, A1 group, A2 group and A3 group There was no statistical significance (P> 0.05). The expressions of α-SMA mRNA, EGFR mRNA and their proteins decreased gradually, with statistical significance (P <0.05). Conclusion EGFR-TKI can inhibit epithelial-mesenchymal transition. Its mechanism may be related to down-regulation of α-SMA and inhibition of EGFR activity. It is speculated that the inhibition of EGFR signaling pathway activation may provide new ideas and ways for the clinical treatment of pulmonary fibrosis.