中国病理生理杂志
中國病理生理雜誌
중국병리생리잡지
CHINESE JOURNAL OF PATHOPHYSIOLOGY
2009年
12期
2403-2407
,共5页
内皮缩血管肽1%气道上皮细胞%成纤维细胞%p38MAPK%PI3K/Akt信号通路
內皮縮血管肽1%氣道上皮細胞%成纖維細胞%p38MAPK%PI3K/Akt信號通路
내피축혈관태1%기도상피세포%성섬유세포%p38MAPK%PI3K/Akt신호통로
Endothelin-1%Airway epithelial cells%Fibroblasts%p38 MAPK%PI3K/Akt signal pathway
目的:探讨p38丝裂原活化蛋白激酶(MAPK)、磷脂酰肌醇3-激酶(PI3K/Akt)在内皮素(ET)-1介导损伤气道上皮诱导上皮下成纤维细胞活化过程中的作用及对白细胞介素(IL)-6的影响.方法:将正常或经多聚左旋精氨酸(PLA)刺激的人气道上皮细胞与原代气道成纤维细胞共培养并分别加入p38 MAPK、PI3K特异性抑制剂SB203580、LY294002或ET受体A阻断剂BQ123,应用免疫组化、免疫印迹技术或ELISA检测成纤维细胞α-平滑肌肌动蛋白(α-SMA)表达、p38 MAPK、Akt的活化及成纤维细胞上清IL-6水平;制备成纤维细胞胶原凝胶并与不同方法处理的上皮细胞共培养,测量各组凝胶面积变化以了解上皮细胞对成纤维细胞收缩反应的诱导及其上述处理因素的影响.结果:与损伤上皮细胞共培养的成纤维细胞上清中ET-1、IL-6水平[(13.69±1.36) ng/L、(56.7±10.7) ng/L]明显高于与正常上皮细胞共培养的成纤维细胞上清[(3.79±0.64) ng/L、(15.5±3.2)ng/L],BQ123、SB203580或LY294002皆不同程度减弱损伤上皮细胞诱导的IL-6释放[分别为(27.2±3.1) ng/L、(31.5±3.6) ng/L、(41.3±3.2) ng/L];成纤维细胞与损伤气道上皮细胞共培养后p38 MAPK、Akt先后激活,BQ123减弱磷酸化p38 MAPK、Akt水平,SB203580浓度依赖性减弱Akt磷酸化水平,而LY294002对磷酸化p38 MAPK水平影响很小.与损伤气道上皮细胞共培养后成纤维细胞α-SMA表达增加,并且胶原收缩百分比明显大于与正常气道上皮共培养的成纤维细胞[(61.2±2.7)% vs (15.4±7.3)%];BQ123、SB203580及LY294002皆不同程度减弱成纤维细胞α-SMA表达与凝胶收缩且BQ123、SB203580抑制凝胶收缩作用较LY294002更明显.结论:ET-1通过激活p38 MAPK、PI3K/Akt信号通路并促进IL-6分泌在损伤气道上皮诱导成纤维细胞活化过程中发挥关键作用.
目的:探討p38絲裂原活化蛋白激酶(MAPK)、燐脂酰肌醇3-激酶(PI3K/Akt)在內皮素(ET)-1介導損傷氣道上皮誘導上皮下成纖維細胞活化過程中的作用及對白細胞介素(IL)-6的影響.方法:將正常或經多聚左鏇精氨痠(PLA)刺激的人氣道上皮細胞與原代氣道成纖維細胞共培養併分彆加入p38 MAPK、PI3K特異性抑製劑SB203580、LY294002或ET受體A阻斷劑BQ123,應用免疫組化、免疫印跡技術或ELISA檢測成纖維細胞α-平滑肌肌動蛋白(α-SMA)錶達、p38 MAPK、Akt的活化及成纖維細胞上清IL-6水平;製備成纖維細胞膠原凝膠併與不同方法處理的上皮細胞共培養,測量各組凝膠麵積變化以瞭解上皮細胞對成纖維細胞收縮反應的誘導及其上述處理因素的影響.結果:與損傷上皮細胞共培養的成纖維細胞上清中ET-1、IL-6水平[(13.69±1.36) ng/L、(56.7±10.7) ng/L]明顯高于與正常上皮細胞共培養的成纖維細胞上清[(3.79±0.64) ng/L、(15.5±3.2)ng/L],BQ123、SB203580或LY294002皆不同程度減弱損傷上皮細胞誘導的IL-6釋放[分彆為(27.2±3.1) ng/L、(31.5±3.6) ng/L、(41.3±3.2) ng/L];成纖維細胞與損傷氣道上皮細胞共培養後p38 MAPK、Akt先後激活,BQ123減弱燐痠化p38 MAPK、Akt水平,SB203580濃度依賴性減弱Akt燐痠化水平,而LY294002對燐痠化p38 MAPK水平影響很小.與損傷氣道上皮細胞共培養後成纖維細胞α-SMA錶達增加,併且膠原收縮百分比明顯大于與正常氣道上皮共培養的成纖維細胞[(61.2±2.7)% vs (15.4±7.3)%];BQ123、SB203580及LY294002皆不同程度減弱成纖維細胞α-SMA錶達與凝膠收縮且BQ123、SB203580抑製凝膠收縮作用較LY294002更明顯.結論:ET-1通過激活p38 MAPK、PI3K/Akt信號通路併促進IL-6分泌在損傷氣道上皮誘導成纖維細胞活化過程中髮揮關鍵作用.
목적:탐토p38사렬원활화단백격매(MAPK)、린지선기순3-격매(PI3K/Akt)재내피소(ET)-1개도손상기도상피유도상피하성섬유세포활화과정중적작용급대백세포개소(IL)-6적영향.방법:장정상혹경다취좌선정안산(PLA)자격적인기도상피세포여원대기도성섬유세포공배양병분별가입p38 MAPK、PI3K특이성억제제SB203580、LY294002혹ET수체A조단제BQ123,응용면역조화、면역인적기술혹ELISA검측성섬유세포α-평활기기동단백(α-SMA)표체、p38 MAPK、Akt적활화급성섬유세포상청IL-6수평;제비성섬유세포효원응효병여불동방법처리적상피세포공배양,측량각조응효면적변화이료해상피세포대성섬유세포수축반응적유도급기상술처리인소적영향.결과:여손상상피세포공배양적성섬유세포상청중ET-1、IL-6수평[(13.69±1.36) ng/L、(56.7±10.7) ng/L]명현고우여정상상피세포공배양적성섬유세포상청[(3.79±0.64) ng/L、(15.5±3.2)ng/L],BQ123、SB203580혹LY294002개불동정도감약손상상피세포유도적IL-6석방[분별위(27.2±3.1) ng/L、(31.5±3.6) ng/L、(41.3±3.2) ng/L];성섬유세포여손상기도상피세포공배양후p38 MAPK、Akt선후격활,BQ123감약린산화p38 MAPK、Akt수평,SB203580농도의뢰성감약Akt린산화수평,이LY294002대린산화p38 MAPK수평영향흔소.여손상기도상피세포공배양후성섬유세포α-SMA표체증가,병차효원수축백분비명현대우여정상기도상피공배양적성섬유세포[(61.2±2.7)% vs (15.4±7.3)%];BQ123、SB203580급LY294002개불동정도감약성섬유세포α-SMA표체여응효수축차BQ123、SB203580억제응효수축작용교LY294002경명현.결론:ET-1통과격활p38 MAPK、PI3K/Akt신호통로병촉진IL-6분비재손상기도상피유도성섬유세포활화과정중발휘관건작용.
AIM: To explore the effects of p38 mitogen-activated protein kinases (MAPK) and phosphoinositide 3-kinases (PI3K)/Akt on interleukin (IL)-6, the endothelin (ET)-1-mediated process of airway fibroblast activation induced by injured human bronchial epithelial cells (HBE). METHODS: Human primary cultured airway fibroblasts were co-cultured with HBE pre-treated with or without poly-L-arginine (PLA). The procedure was also performed in the presence or absence of p38 MAPK selective inhibitor SB203580, PI3K selective inhibitor LY294002 or ETA receptor blocker BQ123, respectively. Immunostaining, Western blotting or ELISA were used for detecting α-smooth muscle actin (α-SMA) expression, the activities of p38 MAPK and Akt in fibroblasts or IL-6 levels in supernatants of fibroblasts. In addition, fibroblasts were mixed with soluble collagen and cultured with HBE treated as the same mentioned above, the gel contraction was measured by serial area measurements. RESULTS: ET-1 and IL-6 levels [(13.69±1.36) ng/L, (56.7±10.7) ng/L] in the supernatants of fibroblasts cultured with injured HBE were significantly higher than those in the supernatants of fibroblasts cultured with HBE [(3.79±0.64) ng/L, (15.5±3.2) ng/L]. BQ123, SB203580 or LY294002 decreased IL-6 levels [(27.2±3.1) ng/L, (31.5±3.6) ng/L, (41.3±3.2) ng/L] differently in the supernatants of fibroblasts induced by injured HBE. Activation of p38 MAPK preceded Akt in fibroblasts cultured with injured HBE. BQ123 reduced the phosphorylation levels of p38 MAPK and Akt. SB203580 concentration-dependently attenuated Akt phosphorylation, while LY294002 had little effect on p38 MAPK phosphorylation. Fibroblasts expressed more α-SMA after cultured with injured HBE and showed significant increase in the gel contraction compared to fibroblasts cultured with HBE [percentage of gel contraction: (61.2±2.7)% vs (15.4±7.3)%], all these effects were diminished or inhibited by BQ123, SB203580 or LY294002. Furthermore, the effects of BQ123 and SB203580 on decreased gel contraction were stronger than the effect of LY294002. CONCLUSION: ET-1 exerts a key role in the airway fibroblasts activation induced by injured HBE through activating p38 MAPK, PI3K/Akt signaling and promoting IL-6 expression.
