中华急诊医学杂志
中華急診醫學雜誌
중화급진의학잡지
CHINESE JOURNAL OF EMERGENCY MEDICINE
2012年
6期
607-611
,共5页
董亮%贺宏丽%刘军%陆晓旻%杨毅%邱海波
董亮%賀宏麗%劉軍%陸曉旻%楊毅%邱海波
동량%하굉려%류군%륙효민%양의%구해파
急性肺损伤%常规树突状细胞%成熟%Th免疫应答%细胞因子%炎症%动态变化
急性肺損傷%常規樹突狀細胞%成熟%Th免疫應答%細胞因子%炎癥%動態變化
급성폐손상%상규수돌상세포%성숙%Th면역응답%세포인자%염증%동태변화
Acute lung injury%Conventional dendritic cells%Maturation%T helper cell immune response%Cytokines%Inflammation%Dynamic change
目的 观察急性肺损伤(ALI)早期肺常规树突状细胞(cDCs)数量及成熟程度的动态改变及其对ALI早期炎症反应和肺损伤的影响.方法 C57BL/6小鼠18只随机(随机数字法)分为健康对照组、6 h-ALI组和24 h-ALI组,气管内注射LPS复制ALI模型后6h及24h分别处死小鼠,留取肺组织,采用流式细胞术检测肺cDCs的数量及成熟程度的动态变化；ELISA检测肺组织IL-6、TNF-α水平评价肺部炎症反应；PCR检测肺组织转录因子T-bet及GATA-3mRNA的表达评价Th1/Th2亚群漂移；ELISA检测肺组织IFN-γ、IL-4水平反映Th1/Th2型细胞因子的平衡；计算肺湿质量/体质量比评价肺水肿程度；肺组织HE染色行组织病理学检查及肺损伤评分反映肺损伤程度.组间比较采用单因素方差分析.结果 ALI组小鼠肺IL-6和TNF-α水平、肺湿质量体质量比及肺损伤评分均较健康对照组显著增高(P＜0.01);6 h-ALI组肺cDCs占肺单个核细胞比例(％)较健康对照组升高[(2.25±0.25)vs(0.93±0.40),P＜0.01],其肺cDCs表达MHC Ⅱ的比例(％)也高于健康对照组[(55.51±11.72) vs.(6.67±0.87),P＜0.01]；而24 h-ALI组肺cDCs占肺单个核细胞比例(％)较6h-ALI组显著降低[(1.01±0.28)vs.(2.25±0.25),P＜0.01],其肺cDCs表达MHC Ⅱ的比例(％)也低于6h-ALI组[(10.94±2.55) vs.(55.51±11.72),P＜0.01]；此外24 h-ALI组肺Th1亚群特异性转录因子T-bet相对表达水平较健康对照组升高[(2.94±0.30)vs(1.00±0.12),P＜0.01],其Th1型细胞因子IFN-γ的水平(Pg·mg-1)也高于健康对照组[(48.17±2.43)vs(16.46±1.68),P＜0.01].结论 ALI早期即存在肺cDCs数量和成熟程度的增加,并可能通过增强Th1型免疫反应及促炎性细胞因子的分泌启动及加剧ALI早期肺部炎症反应.
目的 觀察急性肺損傷(ALI)早期肺常規樹突狀細胞(cDCs)數量及成熟程度的動態改變及其對ALI早期炎癥反應和肺損傷的影響.方法 C57BL/6小鼠18隻隨機(隨機數字法)分為健康對照組、6 h-ALI組和24 h-ALI組,氣管內註射LPS複製ALI模型後6h及24h分彆處死小鼠,留取肺組織,採用流式細胞術檢測肺cDCs的數量及成熟程度的動態變化；ELISA檢測肺組織IL-6、TNF-α水平評價肺部炎癥反應；PCR檢測肺組織轉錄因子T-bet及GATA-3mRNA的錶達評價Th1/Th2亞群漂移；ELISA檢測肺組織IFN-γ、IL-4水平反映Th1/Th2型細胞因子的平衡；計算肺濕質量/體質量比評價肺水腫程度；肺組織HE染色行組織病理學檢查及肺損傷評分反映肺損傷程度.組間比較採用單因素方差分析.結果 ALI組小鼠肺IL-6和TNF-α水平、肺濕質量體質量比及肺損傷評分均較健康對照組顯著增高(P＜0.01);6 h-ALI組肺cDCs佔肺單箇覈細胞比例(％)較健康對照組升高[(2.25±0.25)vs(0.93±0.40),P＜0.01],其肺cDCs錶達MHC Ⅱ的比例(％)也高于健康對照組[(55.51±11.72) vs.(6.67±0.87),P＜0.01]；而24 h-ALI組肺cDCs佔肺單箇覈細胞比例(％)較6h-ALI組顯著降低[(1.01±0.28)vs.(2.25±0.25),P＜0.01],其肺cDCs錶達MHC Ⅱ的比例(％)也低于6h-ALI組[(10.94±2.55) vs.(55.51±11.72),P＜0.01]；此外24 h-ALI組肺Th1亞群特異性轉錄因子T-bet相對錶達水平較健康對照組升高[(2.94±0.30)vs(1.00±0.12),P＜0.01],其Th1型細胞因子IFN-γ的水平(Pg·mg-1)也高于健康對照組[(48.17±2.43)vs(16.46±1.68),P＜0.01].結論 ALI早期即存在肺cDCs數量和成熟程度的增加,併可能通過增彊Th1型免疫反應及促炎性細胞因子的分泌啟動及加劇ALI早期肺部炎癥反應.
목적 관찰급성폐손상(ALI)조기폐상규수돌상세포(cDCs)수량급성숙정도적동태개변급기대ALI조기염증반응화폐손상적영향.방법 C57BL/6소서18지수궤(수궤수자법)분위건강대조조、6 h-ALI조화24 h-ALI조,기관내주사LPS복제ALI모형후6h급24h분별처사소서,류취폐조직,채용류식세포술검측폐cDCs적수량급성숙정도적동태변화；ELISA검측폐조직IL-6、TNF-α수평평개폐부염증반응；PCR검측폐조직전록인자T-bet급GATA-3mRNA적표체평개Th1/Th2아군표이；ELISA검측폐조직IFN-γ、IL-4수평반영Th1/Th2형세포인자적평형；계산폐습질량/체질량비평개폐수종정도；폐조직HE염색행조직병이학검사급폐손상평분반영폐손상정도.조간비교채용단인소방차분석.결과 ALI조소서폐IL-6화TNF-α수평、폐습질량체질량비급폐손상평분균교건강대조조현저증고(P＜0.01);6 h-ALI조폐cDCs점폐단개핵세포비례(％)교건강대조조승고[(2.25±0.25)vs(0.93±0.40),P＜0.01],기폐cDCs표체MHC Ⅱ적비례(％)야고우건강대조조[(55.51±11.72) vs.(6.67±0.87),P＜0.01]；이24 h-ALI조폐cDCs점폐단개핵세포비례(％)교6h-ALI조현저강저[(1.01±0.28)vs.(2.25±0.25),P＜0.01],기폐cDCs표체MHC Ⅱ적비례(％)야저우6h-ALI조[(10.94±2.55) vs.(55.51±11.72),P＜0.01]；차외24 h-ALI조폐Th1아군특이성전록인자T-bet상대표체수평교건강대조조승고[(2.94±0.30)vs(1.00±0.12),P＜0.01],기Th1형세포인자IFN-γ적수평(Pg·mg-1)야고우건강대조조[(48.17±2.43)vs(16.46±1.68),P＜0.01].결론 ALI조기즉존재폐cDCs수량화성숙정도적증가,병가능통과증강Th1형면역반응급촉염성세포인자적분비계동급가극ALI조기폐부염증반응.
Objective To investigate the accumulation and maturation status of pulmonary conventional dendritic cells (cDCs) in the early phase of acute lung injury (ALI),and to explore the way of the inflammatory responses and lung injury modulated by cDCs in vivo.Methods Male C57BL/6 mice were randomly ( random number) divided into the normal control group,6 h-ALI group and 24 h-ALI group.Murine model of ALI was made by intra-tracheal administration of lipopolysaccharide (LPS) and lung specimens were taken 6 h or 24 h later.The accumulation and maturation status of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate the lung inflammation.Transcription factors T-bet/GATA-3 mRNA ratio was determined to estimate the balance between Th1/Th2 responses.IFN-γand IL-4 were quantified to evaluate Thl-specific and Th2-specific cytokine production respectively.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Comparison between groups was performed using one -way ANOVA.Results Compared with normal control group,LPS challenge resulted in higher level of IL-6 and TNF-α,increased LWW/BW ratio and significant histopathological changes (P ＜0.01 ).The accumulation and maturation of pulmonary cDCs in 6 h-ALI group were significantly increased after LPS challenge (P ＜0.01 ),while the accumulation and maturation of pulmonary cDCs in 24 h-ALI group were significantly lower than that in 6 h-ALI group ( P ＜0.01 ).Compared with normal control group,the expression of T-bet mRNA in 24 h-ALI group was markedly enhanced ( P ＜ 0.01 ) and the production of IFN-γ was increased as well ( P ＜ 0.01 ).Conclusions The accumulation and maturation of pulmonary cDCs peaked within 24 h after LPS challenge,pulmonary cDCs may initiate and amplify acute lung inflammation of ALI by enhancing the Th1 immune response and ensuing cytokine production.
