CAJ | 학술논문

目的探讨应用吉非替尼抑制表皮生长因子受体(EGFR)酪氨酸激酶活性对慢性阻塞性肺疾病(COPD)黏液高分泌的影响. 方法采用香烟提取物(CSE)刺激支气管上皮细胞株16HBE制备COPD模型,采用EGFR酪氨酸激酶抑制剂吉非替尼抑制模型细胞的EGFR自身酪氨酸位点磷酸化,实时定量聚合酶链反应(real-time PCR)检测EGFR、黏蛋白MUC5 AC基因水平,Western blot检测EGFR、磷酸化EGFR(p EGFR)蛋白表达水平,酶联免疫吸附法(ELISA)检测MUC5AC蛋白水平. 结果与对照组比较,CSE组、吉非替尼组的EGFR基因表达分别上调12.7％、8.6％,差异均无统计学意义(P＞0.05)；MUC5AC基因水平分别上调141.7％、26.4％,差异有统计学意义(P＜0.05).对照组、CSE组、吉非替尼组EGFR蛋白相对含量分别为600.34±64.58、632.58±72.94、584.57±67.39,各组之间比较差异无统计学意义(P＞0.05).p-EGFR蛋白相对含量分别为338.62±45.28、679.43±78.23、292.74±59.17,CSE组p-EGFR蛋白含量较对照组显著增加(P＜0.05),吉非替尼组p EGFR蛋白水平与对照组比较差异无统计学意义(P＞0.05).对照组、CSE组、吉非替尼组MUC5AC蛋白含量分别为(72.80±6.25)μg/mg、(187.00±10.26) μg/mg,(92.57±8.32)μg/mg,与对照组比较,CSE组MUC5AC蛋白表达显著增加(P＜0.05),吉非替尼组MUC5AC蛋白水平与对照组比较差异无统计学意义(P＞0.05). 结论香烟烟雾可能通过EGFR途径导致气道黏液过度分泌,应用EGFR受体酪氨酸激酶抑制剂吉非替尼可通过抑制EGFR自身酪氨酸激酶活性,有效阻断香烟烟雾诱发的气道黏液高分泌,EGFR作为COPD治疗的一个靶点可行有效.
목적 탐토응용길비체니억제표피생장인자수체(EGFR)락안산격매활성대만성조새성폐질병(COPD)점액고분비적영향. 방법 채용향연제취물(CSE)자격지기관상피세포주16HBE제비COPD모형,채용EGFR락안산격매억제제길비체니억제모형세포적EGFR자신락안산위점린산화,실시정량취합매련반응(real-time PCR)검측EGFR、점단백MUC5 AC기인수평,Western blot검측EGFR、린산화EGFR(p EGFR)단백표체수평,매련면역흡부법(ELISA)검측MUC5AC단백수평. 결과 여대조조비교,CSE조、길비체니조적EGFR기인표체분별상조12.7％、8.6％,차이균무통계학의의(P＞0.05)；MUC5AC기인수평분별상조141.7％、26.4％,차이유통계학의의(P＜0.05).대조조、CSE조、길비체니조EGFR단백상대함량분별위600.34±64.58、632.58±72.94、584.57±67.39,각조지간비교차이무통계학의의(P＞0.05).p-EGFR단백상대함량분별위338.62±45.28、679.43±78.23、292.74±59.17,CSE조p-EGFR단백함량교대조조현저증가(P＜0.05),길비체니조p EGFR단백수평여대조조비교차이무통계학의의(P＞0.05).대조조、CSE조、길비체니조MUC5AC단백함량분별위(72.80±6.25)μg/mg、(187.00±10.26) μg/mg,(92.57±8.32)μg/mg,여대조조비교,CSE조MUC5AC단백표체현저증가(P＜0.05),길비체니조MUC5AC단백수평여대조조비교차이무통계학의의(P＞0.05). 결론 향연연무가능통과EGFR도경도치기도점액과도분비,응용EGFR수체락안산격매억제제길비체니가통과억제EGFR자신락안산격매활성,유효조단향연연무유발적기도점액고분비,EGFR작위COPD치료적일개파점가행유효.
Objective To investigate the effect of gefitinib on mucus hypersecretion by inhibiting epidermal growth factor receptor (EGFR) activity in chronic obstructive pulmonary disease (COPD).Methods Human airway epithelail cell lines 16HBE cells were exposed to cigarette smoke extraction (CSE) to establish the COPD model.EGFR activity was inhibited by tyrosine kinase inhibitor gefitinib.The mRNA expressions of EGFR and MUC5AC were detected by real-time PCR.EGFR,p-EGFR and MUC5AC protein levels were determined by Western blot and ELISA.Results EGFR mRNA level was increased by 12.7％ in CSE and 8.6％ in gefitinib group,but had no significant differences among CSE,gefitinib group and control group (all P＞ 0.05).MUC5AC mRNA levels were enhanced by 141.7％,26.4％ in CSE group and gefitinib group respectively,and there were significant differences among CSE,gefitinib group and control group (all P＜0.05).EGFR protein levels were (600.34±64.58) μg/mg,(632.58±72.94) μg/mg,(584.57±67.39) μg/mg,in control,CSE and gefitinib groups,respectively,and there were no significant differences between groups (all P＞0.05).p-EGFR protein levels were (338.62±45.28) μg/mg,(679.43±78.23) μg/mg,(292.74±59.17) μg/mg in control,CSE and gefitinib groups,respectively.MUC5AC protein levels were(72.80±6.25)μg/mg,(187.00±±10.26)μg/mg,(92.57±8.32)μg/mg in control,CSE and gefitinib groups respectively.Compared with control group,p-EGFR and MUC5AC protein levels were increased significantly in CSE group (both P＜0.05),and had no significant differences in p EGFR and MUC5AC protein levels between control group and gefitinib group.Conclusions CSE may lead to mucus hypersecretion through activating the EGFR-mediated signaling pathways.Gefitinib may inhibit mucus hypersecretion by inhibiting EGFR tyrosine kinanse activity.EGFR may serve as a potential target for COPD.