中华器官移植杂志
中華器官移植雜誌
중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2014年
9期
561-566
,共6页
周志毅%朱幸沨%陈静瑜%郭庆%杨国仪
週誌毅%硃倖沨%陳靜瑜%郭慶%楊國儀
주지의%주행풍%진정유%곽경%양국의
大鼠%肺%再灌注损伤%TLR4%缺氧诱导因子-1α
大鼠%肺%再灌註損傷%TLR4%缺氧誘導因子-1α
대서%폐%재관주손상%TLR4%결양유도인자-1α
Rats%Lung%Reperfusion injury%Toll-like receptor 4%Hypoxia-inducible factor-1α
目的 观察大鼠肺缺血再灌注损伤(IRI)模型中TLR4信号通路与HIF-1α的相互关系及影响,探讨HIF-1α表达在TLR4信号通路介导的肺IRI中的影响及机制.方法 45只雄性SD大鼠,分为9组进行观察.(1)假手术组;(2)肺IRI组;(3)肺IRI+生理盐水对照组;(4)肺IRI+ DMSO对照组;(5)肺IRI+脂多糖干预组(TLR4激活组);(6)肺IRI+ TAK-242干预组(TLR4抑制组);(7)肺IRI+硫氧还蛋白干预组(ASK1抑制组);(8)肺IRI+ SB203580干预组(P38抑制组);(9)肺IRI+毛壳菌素干预组(HIF-1α抑制组).制作大鼠原位肺IRI模型,取各组大鼠左肺组织,观察肺组织病理变化、行逆转录-聚合酶链反应及蛋白质印迹法检测,统计分析TLR4信号通路(TLR4、MyD88、TRIF、ASK1及p38)与HIF-1α的相互关系及作用.结果 脂多糖干预组HIF-1α mRNA及蛋白的表达较TAK-242干预组明显增加(P＜0.05);MyD88、P-ASK1及P-p38的表达较TAK-242干预组明显增强(P＜0.05);经人重组硫氧还蛋白特异性抑制ASK1,肺IRI组中P-p38及HIF-1α表达明显减少(P＜0.05),经p38特异性抑制剂SB203580干预,肺IRI组HIF-1α的表达同样受到显著抑制(P＜0.05),而各组总ASK1及p38无明显变化,TRIF的表达则亦未受到明显影响.对比肺IRI组,应用特异性HIF-1α抑制剂毛壳菌素可显著减轻肺组织TNF-α和IL-6的表达,减轻细胞凋亡及肺损伤评分(P＜0.05).TLR4 mRNA和蛋白的表达被毛壳菌素显著抑制(P＜0.05).结论 HIF-1α在TLR4信号通路介导的肺IRI中具有损害性作用,TLR4和HIF-1α可能协同参与肺IRI的发生、发展,其可能是治疗肺IRI新的有效靶点.
目的 觀察大鼠肺缺血再灌註損傷(IRI)模型中TLR4信號通路與HIF-1α的相互關繫及影響,探討HIF-1α錶達在TLR4信號通路介導的肺IRI中的影響及機製.方法 45隻雄性SD大鼠,分為9組進行觀察.(1)假手術組;(2)肺IRI組;(3)肺IRI+生理鹽水對照組;(4)肺IRI+ DMSO對照組;(5)肺IRI+脂多糖榦預組(TLR4激活組);(6)肺IRI+ TAK-242榦預組(TLR4抑製組);(7)肺IRI+硫氧還蛋白榦預組(ASK1抑製組);(8)肺IRI+ SB203580榦預組(P38抑製組);(9)肺IRI+毛殼菌素榦預組(HIF-1α抑製組).製作大鼠原位肺IRI模型,取各組大鼠左肺組織,觀察肺組織病理變化、行逆轉錄-聚閤酶鏈反應及蛋白質印跡法檢測,統計分析TLR4信號通路(TLR4、MyD88、TRIF、ASK1及p38)與HIF-1α的相互關繫及作用.結果 脂多糖榦預組HIF-1α mRNA及蛋白的錶達較TAK-242榦預組明顯增加(P＜0.05);MyD88、P-ASK1及P-p38的錶達較TAK-242榦預組明顯增彊(P＜0.05);經人重組硫氧還蛋白特異性抑製ASK1,肺IRI組中P-p38及HIF-1α錶達明顯減少(P＜0.05),經p38特異性抑製劑SB203580榦預,肺IRI組HIF-1α的錶達同樣受到顯著抑製(P＜0.05),而各組總ASK1及p38無明顯變化,TRIF的錶達則亦未受到明顯影響.對比肺IRI組,應用特異性HIF-1α抑製劑毛殼菌素可顯著減輕肺組織TNF-α和IL-6的錶達,減輕細胞凋亡及肺損傷評分(P＜0.05).TLR4 mRNA和蛋白的錶達被毛殼菌素顯著抑製(P＜0.05).結論 HIF-1α在TLR4信號通路介導的肺IRI中具有損害性作用,TLR4和HIF-1α可能協同參與肺IRI的髮生、髮展,其可能是治療肺IRI新的有效靶點.
목적 관찰대서폐결혈재관주손상(IRI)모형중TLR4신호통로여HIF-1α적상호관계급영향,탐토HIF-1α표체재TLR4신호통로개도적폐IRI중적영향급궤제.방법 45지웅성SD대서,분위9조진행관찰.(1)가수술조;(2)폐IRI조;(3)폐IRI+생리염수대조조;(4)폐IRI+ DMSO대조조;(5)폐IRI+지다당간예조(TLR4격활조);(6)폐IRI+ TAK-242간예조(TLR4억제조);(7)폐IRI+류양환단백간예조(ASK1억제조);(8)폐IRI+ SB203580간예조(P38억제조);(9)폐IRI+모각균소간예조(HIF-1α억제조).제작대서원위폐IRI모형,취각조대서좌폐조직,관찰폐조직병리변화、행역전록-취합매련반응급단백질인적법검측,통계분석TLR4신호통로(TLR4、MyD88、TRIF、ASK1급p38)여HIF-1α적상호관계급작용.결과 지다당간예조HIF-1α mRNA급단백적표체교TAK-242간예조명현증가(P＜0.05);MyD88、P-ASK1급P-p38적표체교TAK-242간예조명현증강(P＜0.05);경인중조류양환단백특이성억제ASK1,폐IRI조중P-p38급HIF-1α표체명현감소(P＜0.05),경p38특이성억제제SB203580간예,폐IRI조HIF-1α적표체동양수도현저억제(P＜0.05),이각조총ASK1급p38무명현변화,TRIF적표체칙역미수도명현영향.대비폐IRI조,응용특이성HIF-1α억제제모각균소가현저감경폐조직TNF-α화IL-6적표체,감경세포조망급폐손상평분(P＜0.05).TLR4 mRNA화단백적표체피모각균소현저억제(P＜0.05).결론 HIF-1α재TLR4신호통로개도적폐IRI중구유손해성작용,TLR4화HIF-1α가능협동삼여폐IRI적발생、발전,기가능시치료폐IRI신적유효파점.
Objective To investigate the effect of hypoxia-inducible factor-1α expression (HIF-1α) on toll-like receptor 4 (TLR4) signaling pathway-mediated rat lung ischemia-reperfusion injury (LIRI).Method Forty-five S-D rats were randomly divided into Sham group,LIRI group,LIRI+ TLR4-activated group,LIRI+ TLR4-inhibited group,LIRI + ASK1-inhibited group,LIRI + p38-inhibited group,and LIRI + HIF-1α-inhibited group.The interaction between TLR4 signaling pathway [including TLR4,myeloid differentiation factor 88 (MyD88),TIR-domain-containing adapter-inducing interferon-βTIR-domain-containing adapter-inducing interferon-β (TRIF),Apoptosis signal-regulating kinase 1 (ASK1) and p38] and HIF-1α and the role of TLR4-dependent HIF-1α in LIRI in vivo were analyzed.Result In LIRI,HIF-1α accumulation was induced in a TLR4-dependent fashion,and MyD88,but not TRIF,and activation of ASK1 and P38 were found to be critical for TLR4-mediated HIF-1α accumulation.HIF-1α protein played a critical role in TLR4-mediated lung injury of LIRI.HIF-1α up-regulated TLR4 expression in LIRI in a positive feedback manner.Conclusion We identify that HIF-1α has a damaging effect on TLR4 signaling pathway-mediated LIRI and TLR4-HIF-1 may synergistically involved in the development of LIRI.Therefore we suggest that the interaction between them may represent a novel therapeutic target for the development of novel target-based therapies of LIRI.
