中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
28期
1988-1991
,共4页
周佳%尤列·皮尔曼%维克多·科罗索夫%周向东
週佳%尤列·皮爾曼%維剋多·科囉索伕%週嚮東
주가%우렬·피이만%유극다·과라색부%주향동
神经调节蛋白1%白细胞介素1β%黏蛋白类%受体%表皮生长因子
神經調節蛋白1%白細胞介素1β%黏蛋白類%受體%錶皮生長因子
신경조절단백1%백세포개소1β%점단백류%수체%표피생장인자
Neuregulin-1%Interleukin-1beta%Mucins%Receptor%epidermal growth factor
目的 探讨神经调节蛋白1β(NRG-1β)在白细胞介素1β(IL-1β)诱导的气道黏液高分泌中的作用.方法 用IL-1β刺激人气道上皮细胞株HBE16细胞构建黏液高分泌模型,逆转录PCR法检测NRG-1β mRNA和黏蛋白(MUC)5AC mRNA,ELISA法检测NRG-1β和MUC5AC蛋白水平.用NRG-1β刺激后,ELISA法检测MUC5AC表达,Western印迹法检测磷酸化人表皮生长因子受体(ErbB)1 ～4.预先分别给予ErbB1～4抗体及p38触分裂原活化蛋白(MAPK)特异性抑制剂、细胞外信号调节蛋白(ERK)1/2特异性抑制剂、丝裂原压力活性蛋白激酶(MSK)1特异性抑制剂、环磷酸腺苷(c-AMP)反应原件结合蛋白(CREB)单克隆抗体,再经NRG-1β刺激后,ELISA法检测MUC5 AC蛋白表达.结果 IL-1β显著增加NRG-1β、MUC5AC mRNA水平及其蛋白表达,且在蛋白水平呈现浓度正相关性.单独给予NRG-1β(1、10、100、200 nmol/L)刺激HBE16细胞,MUC5 AC蛋白表达(0.328±0.055、0.364±0.086、0.650±0.134、0.586±0.068)较对照组(0.227 ±0.019)显著增加,差异均有统计学意义(均P＜0.05),同时磷酸化ErbB2、3呈阳性表达,而磷酸化ErbB1、4呈近似阴性表达.预先给予ErbB2、3抗体、p38MAPK特异性抑制剂、ERK1/2特异性抑制剂和MSK1抑制剂、CREB抗体再给予NRG-1β刺激者MUC5AC蛋白表达(0.221±0.033、0.238±0.044、0.386±0.021、0.352 ±0.022、0.294±0.017、0.252±0.019)较单独给予NRG-1β(0.650±0.134)显著降低,差异均有统计学意义(均P＜0.05).结论 IL-1β导致气道黏液高分泌可能通过NRG- 1β/ErbB2、3异二聚体信号通路,并激活后续的MAPK/MSK1/CREB信号路径实现.
目的 探討神經調節蛋白1β(NRG-1β)在白細胞介素1β(IL-1β)誘導的氣道黏液高分泌中的作用.方法 用IL-1β刺激人氣道上皮細胞株HBE16細胞構建黏液高分泌模型,逆轉錄PCR法檢測NRG-1β mRNA和黏蛋白(MUC)5AC mRNA,ELISA法檢測NRG-1β和MUC5AC蛋白水平.用NRG-1β刺激後,ELISA法檢測MUC5AC錶達,Western印跡法檢測燐痠化人錶皮生長因子受體(ErbB)1 ～4.預先分彆給予ErbB1～4抗體及p38觸分裂原活化蛋白(MAPK)特異性抑製劑、細胞外信號調節蛋白(ERK)1/2特異性抑製劑、絲裂原壓力活性蛋白激酶(MSK)1特異性抑製劑、環燐痠腺苷(c-AMP)反應原件結閤蛋白(CREB)單剋隆抗體,再經NRG-1β刺激後,ELISA法檢測MUC5 AC蛋白錶達.結果 IL-1β顯著增加NRG-1β、MUC5AC mRNA水平及其蛋白錶達,且在蛋白水平呈現濃度正相關性.單獨給予NRG-1β(1、10、100、200 nmol/L)刺激HBE16細胞,MUC5 AC蛋白錶達(0.328±0.055、0.364±0.086、0.650±0.134、0.586±0.068)較對照組(0.227 ±0.019)顯著增加,差異均有統計學意義(均P＜0.05),同時燐痠化ErbB2、3呈暘性錶達,而燐痠化ErbB1、4呈近似陰性錶達.預先給予ErbB2、3抗體、p38MAPK特異性抑製劑、ERK1/2特異性抑製劑和MSK1抑製劑、CREB抗體再給予NRG-1β刺激者MUC5AC蛋白錶達(0.221±0.033、0.238±0.044、0.386±0.021、0.352 ±0.022、0.294±0.017、0.252±0.019)較單獨給予NRG-1β(0.650±0.134)顯著降低,差異均有統計學意義(均P＜0.05).結論 IL-1β導緻氣道黏液高分泌可能通過NRG- 1β/ErbB2、3異二聚體信號通路,併激活後續的MAPK/MSK1/CREB信號路徑實現.
목적 탐토신경조절단백1β(NRG-1β)재백세포개소1β(IL-1β)유도적기도점액고분비중적작용.방법 용IL-1β자격인기도상피세포주HBE16세포구건점액고분비모형,역전록PCR법검측NRG-1β mRNA화점단백(MUC)5AC mRNA,ELISA법검측NRG-1β화MUC5AC단백수평.용NRG-1β자격후,ELISA법검측MUC5AC표체,Western인적법검측린산화인표피생장인자수체(ErbB)1 ～4.예선분별급여ErbB1～4항체급p38촉분렬원활화단백(MAPK)특이성억제제、세포외신호조절단백(ERK)1/2특이성억제제、사렬원압력활성단백격매(MSK)1특이성억제제、배린산선감(c-AMP)반응원건결합단백(CREB)단극륭항체,재경NRG-1β자격후,ELISA법검측MUC5 AC단백표체.결과 IL-1β현저증가NRG-1β、MUC5AC mRNA수평급기단백표체,차재단백수평정현농도정상관성.단독급여NRG-1β(1、10、100、200 nmol/L)자격HBE16세포,MUC5 AC단백표체(0.328±0.055、0.364±0.086、0.650±0.134、0.586±0.068)교대조조(0.227 ±0.019)현저증가,차이균유통계학의의(균P＜0.05),동시린산화ErbB2、3정양성표체,이린산화ErbB1、4정근사음성표체.예선급여ErbB2、3항체、p38MAPK특이성억제제、ERK1/2특이성억제제화MSK1억제제、CREB항체재급여NRG-1β자격자MUC5AC단백표체(0.221±0.033、0.238±0.044、0.386±0.021、0.352 ±0.022、0.294±0.017、0.252±0.019)교단독급여NRG-1β(0.650±0.134)현저강저,차이균유통계학의의(균P＜0.05).결론 IL-1β도치기도점액고분비가능통과NRG- 1β/ErbB2、3이이취체신호통로,병격활후속적MAPK/MSK1/CREB신호로경실현.
Objective To explore the role of neuregulin 1β (NRG-1β) in airway hypersecretion induced by interleukin ( IL)-1β.Methods After stimulating the airway epithelial cell line HBE16 with IL-1β,the expressions of NRG-1β mRNA and mucin (MUC) 5AC mRNA were detected by reverse transcription-polymerese chain reaction (RT-PCR),the proteins of NRG-1β and MUC5AC measured by enzyme-linked immunnsorbent assay ( ELISA ) and phosphorylated erythroblastic leukemia viral oncogene homolog (ErbB)1 -4 detected by Western blot.The cells were pre-treated with antibodies of ErbBI -4,inhibitors of p38 mitogen-activated protein kinase ( MAPK),ERK1/2,mitogen- and stress-activated protein kinase (MSK)1 and antibody of cAMP-response element-binding protein (CREB).After the addition of stimulant NRG-1β,MUCSAC was measured by ELISA.Results IL-1β could increase markedly the levels of NRG-1 β mRNA and MUC5AC mRNA and also the proteins of NRG-1β and MUC5AC in a dose-dependent fashion.NRG-1β at concentrations of 1,10,100,200 nmol/L increased the expression of MUC5AC (0.328±0.055,0.364 ±0.086,0.650 ±0.134,0.586 ±0.068) versus the control group (0.227 ±0.019 ).And the results had statistical significances (P ＜ 0.05 ).The expressions of phosphorylated ErbB2 and ErbB3 stimulated by NRG-1 β were positive while those of phosphorylated ErbB1 and ErbB 4 negative.After a pretreatment of antibodies of ErbB2,ErbB3,inhibitors of p38MAPK,ERK1/2,MSK1 and antibody of CREB and a stimulation of NRG-1β,the expression of MUC5AC decreased (0.221 ±0.033,0.238 ±0.044,0.386 ±0.021,0.352 ±0.022,0.294 ±0.017,0.252 ±0.019) versus the NRG-1β group (0.650 ± 0.134).And the results had statistical significances ( P ＜ 0.05 ).Conclusion IL-1 β may cause airway hypersecreton probably through the combination of NRG-1β with ErbB2 and ErbB3 heterodimers and the activation of MAPK/MSK1/CREB signal conduction.
