基础医学与临床
基礎醫學與臨床
기출의학여림상
BASIC MEDICAL SCIENCES AND CLINICS
2010年
3期
246-251
,共6页
黏蛋白类%香烟烟雾%受体%表皮生长因子%血红素加氧酶-1
黏蛋白類%香煙煙霧%受體%錶皮生長因子%血紅素加氧酶-1
점단백류%향연연무%수체%표피생장인자%혈홍소가양매-1
mucins%cigarette smoke%receptor%epidermal growth factor%heine oxygenase-1
目的 探讨血红素加氧酶-1(HO-1)对香烟烟雾提取物(CSE)所致人气道黏液高分泌的影响.方法 用CSE刺激A549细胞,复制黏液高分泌细胞模型.分为对照组、CSE组、氯化高铁血红素(Heroin)组和锌原卟啉(ZnPPIX)组,观察黏蛋白(MUC)5AC、表皮生长因子受体(EGFR)、磷酸化(P)-EGFR、HO-1及双功能氧化酶1(Duoxl)的表达.用MTT法测定细胞活性;RT-PCR法检测HO-1、Duox1、EGFR及MUC5AC的mRNA;Western blot法检测P-EGFR、EGFR、Duox1和HO-1蛋白;ELISA法检测细胞裂解液中MUCSAC蛋白表达.结果 CSE组MUC5AC的mRNA吸光度积分相对值和蛋白水平分别为0.660±0.044和(157±3)μg/mg,较对照组0.412±0.043和(105±8)μg/mg明显升高(P＜0.05),p-EGFR蛋白水平、EGFR、HO-1及Duox1的mRNA和蛋白水平也较对照组明显增高.与CSE组相比,Hemin组140-1 mRNA及蛋白进一步增高,而p-EGFR蛋白水平、Duox1、EGFR及MUCSAC的mRNA和蛋白水平明显回降.与对照组相比,ZnPPIX组HO-1 mRNA及蛋白水平无明显增高,而p-EGFR蛋白水平、Duox1、EGFR及MUC5AC的mRNA和蛋白水平明显增高.结论 HO-1可通过下调Duox1的表达,阻断香烟烟雾诱导EGFR活化的上游信号途径,减少EGFR活化,从而抑制MUCSAC的表达.
目的 探討血紅素加氧酶-1(HO-1)對香煙煙霧提取物(CSE)所緻人氣道黏液高分泌的影響.方法 用CSE刺激A549細胞,複製黏液高分泌細胞模型.分為對照組、CSE組、氯化高鐵血紅素(Heroin)組和鋅原卟啉(ZnPPIX)組,觀察黏蛋白(MUC)5AC、錶皮生長因子受體(EGFR)、燐痠化(P)-EGFR、HO-1及雙功能氧化酶1(Duoxl)的錶達.用MTT法測定細胞活性;RT-PCR法檢測HO-1、Duox1、EGFR及MUC5AC的mRNA;Western blot法檢測P-EGFR、EGFR、Duox1和HO-1蛋白;ELISA法檢測細胞裂解液中MUCSAC蛋白錶達.結果 CSE組MUC5AC的mRNA吸光度積分相對值和蛋白水平分彆為0.660±0.044和(157±3)μg/mg,較對照組0.412±0.043和(105±8)μg/mg明顯升高(P＜0.05),p-EGFR蛋白水平、EGFR、HO-1及Duox1的mRNA和蛋白水平也較對照組明顯增高.與CSE組相比,Hemin組140-1 mRNA及蛋白進一步增高,而p-EGFR蛋白水平、Duox1、EGFR及MUCSAC的mRNA和蛋白水平明顯迴降.與對照組相比,ZnPPIX組HO-1 mRNA及蛋白水平無明顯增高,而p-EGFR蛋白水平、Duox1、EGFR及MUC5AC的mRNA和蛋白水平明顯增高.結論 HO-1可通過下調Duox1的錶達,阻斷香煙煙霧誘導EGFR活化的上遊信號途徑,減少EGFR活化,從而抑製MUCSAC的錶達.
목적 탐토혈홍소가양매-1(HO-1)대향연연무제취물(CSE)소치인기도점액고분비적영향.방법 용CSE자격A549세포,복제점액고분비세포모형.분위대조조、CSE조、록화고철혈홍소(Heroin)조화자원계람(ZnPPIX)조,관찰점단백(MUC)5AC、표피생장인자수체(EGFR)、린산화(P)-EGFR、HO-1급쌍공능양화매1(Duoxl)적표체.용MTT법측정세포활성;RT-PCR법검측HO-1、Duox1、EGFR급MUC5AC적mRNA;Western blot법검측P-EGFR、EGFR、Duox1화HO-1단백;ELISA법검측세포렬해액중MUCSAC단백표체.결과 CSE조MUC5AC적mRNA흡광도적분상대치화단백수평분별위0.660±0.044화(157±3)μg/mg,교대조조0.412±0.043화(105±8)μg/mg명현승고(P＜0.05),p-EGFR단백수평、EGFR、HO-1급Duox1적mRNA화단백수평야교대조조명현증고.여CSE조상비,Hemin조140-1 mRNA급단백진일보증고,이p-EGFR단백수평、Duox1、EGFR급MUCSAC적mRNA화단백수평명현회강.여대조조상비,ZnPPIX조HO-1 mRNA급단백수평무명현증고,이p-EGFR단백수평、Duox1、EGFR급MUC5AC적mRNA화단백수평명현증고.결론 HO-1가통과하조Duox1적표체,조단향연연무유도EGFR활화적상유신호도경,감소EGFR활화,종이억제MUCSAC적표체.
Objective To investigate the effects of heme oxygenase-1 (HO-1) on the signal transduction pathway of mucin(MUC) expression induced by cigarette smoke.Methods The cell model of mucous hypersecretion was made by human lung A549 cell stimulated by cigarette smoke extract(CSE),the cells were divided into 4 groups:negative control group,CSE treatment group,heroin pre-treatment group and ZnPPIX pre-treatment group.The expression of MUC5AC,epidermal growth factor receptor(EGFR),p-EGFR,HO-1 and dual oxidase 1 (Duoxl) were detected.The cell activity was assessed by methyl thiazolyl tetrazolium method.The change of HO-1 mRNA,Duoxl mRNA,EGFR mRNA and MUC5AC mRNA was examined by reverse transcriptase-polymerase chain reaction.The protein expression of EGFR,p-EGFR,Duox1 and HO-1 was measured by Western blot,while the pro tein expression changes of MUC 5 AC were detected by ELISA.Results The expression level of MUC 5 AC mRNA and its protein in the CSE group increased significantly (P＜0.05) as compared to those in the control group [0.412±0.043,(105±8) μg/mg.The mRNA and protein of EGFR,Duox1 and HO-1,the protein of p-EGFR increased significantly as compared to the control group.After the cells being pre-treated with hemin,the mRNA and protein of HO-1 increased significantly,while the mRNA and protein of Duox1,EGFR and MUC,SAC,the protein of p-EGFR decreased significantly as compared to the CSE group.After the cells were pre-treated with ZnPPIX,the increase of HO-1 mRNA and protein was not significant as compared to the control group,while the mRNA and protein of Duox1,EGFR and MUCSAC,the protein of p-EGFR increased significantly.Conclusion HO-1 decreased the level of Duox1,blocked ligand-dependent EGF-R activation and decreased the expression levels of MUCSAC.
