中华结核和呼吸杂志
中華結覈和呼吸雜誌
중화결핵화호흡잡지
Chinese Journal of Tuberculosis and Respiratory Diseases
2011年
4期
259-264
,共6页
段敏超%钟小宁%何志义%唐海娟%黄颖
段敏超%鐘小寧%何誌義%唐海娟%黃穎
단민초%종소저%하지의%당해연%황영
T淋巴细胞,辅助诱导%白细胞介素类%烟草烟污染%肺炎%肺气肿
T淋巴細胞,輔助誘導%白細胞介素類%煙草煙汙染%肺炎%肺氣腫
T림파세포,보조유도%백세포개소류%연초연오염%폐염%폐기종
T-lymphocytes,helper-inducer%Interleukins%Tobacco smoke pollution%Pneumonia%Pulmonary emphysema
目的 观察香烟烟雾暴露小鼠肺实质中CD4+白细胞介素(IL)-17+辅助性T细胞(Th17)数量及活性的表达,探讨其在香烟暴露小鼠肺部CD4+ γ-干扰素+(Th1)炎症及肺气肿中的作用及相关机制.方法 将40只雄性Balb/c小鼠按随机数字表法分为4组:对照12周(C12)组、对照24周(C24)组、烟雾暴露12周(S12)组、烟雾暴露24周(S24)组,每组10只.香烟烟雾暴露法建立小鼠肺气肿模型.HE染色观察小鼠肺气肿的改变,计算平均内衬间隔和肺泡破坏指数(DI);流式细胞术检测小鼠肺实质中CD4+IL-17+T(Th17)细胞、CD4+γ-干扰素+T(Th1)细胞、CD4+IL-17+γ-干扰素+T(Th17/Th1)细胞、CD8+γ-干扰素+T(Tc1)细胞、CD8+IL-21R+细胞及CD4+IL-17+IL-21+细胞比例;荧光定量PCR法检测小鼠肺实质中维甲酸相关孤独受体(RORγt)和IL-17的mRNA表达,并分析这些指标的相互关系.结果 S12组和S24组的平均内衬间隔[(39±4)μm和(47±7)μm]和DI(39.1±1.6和45.2±3.1)明显高于C12组[(32±4)μm和28.2±1.6]和C24组[(33±3)μm和28.9±2.1],且以S24组的增高更为明显,差异均有统计学意义(t值为4.378～15.188,均P＜0.05);S12组和S24组Th17细胞比例[(3.3±1.1)%和(7.2±2.2)%]均明显高于C12组和C24组[(1.8±0.8)%和(2.0±0.6)%];S12组和S24组RORγtmRNA表达量[(25±4)和(35±3)]及IL-17的mRNA表达量[(26±3)和(36±3)]亦明显高于C12组[(10±5)和(13±5)]和C24组[(11±7)和(8±6)],以S24组增高更为明显,差异均有统计学意义(P＜0.05);S12组和S24组Th1细胞比例[(10.0±3.7)%和(26.2 ±6.0)%]、Th17/Th1细胞比例[(0.61±0.30)%和(1.82±0.52)%]及Tc1细胞比例[(17.0±4.5)%和(26.8±8.5)%]均明显高于C12组[(3.8±1.7)%、(0.27±0.17)%和(4.8±1.9)%]和C24组[(4.2±1.3)%、(0.28±0.11)%和(5.2±1.0)%],以S24组增高更为明显,差异均有统计学意义(P＜0.05);S12组和S24组小鼠Th17细胞与Th1、Tc1细胞比例、平均内衬间隔、DI值均呈显著正相关(r值为0.519～0.797,均P＜0.01);Th17/Th1细胞比例与平均内衬间隔、DI值呈显著正相关(r值分别为0.742和0.802,均P＜0.01);S12组和S24组CD4+IL-17+IL-21+细胞比例[(0.19±0.04)和(0.55±0.24)]明显高于C12组和C24组[(0.07±0.03)和(0.08±0.03)],S24组增高更为明显,差异均有统计学意义(P＜0.05).S12组和S24组的CD8+IL-21R+细胞比例[(2.94±1.26)和(4.12±2.26)]高于C12组和C24组[(1.22±0.31)和(1.34±0.18)](P＞0.05);S12组及S24组小鼠CD4+IL-17+IL-21+细胞比例与Th1、Tc1细胞比例、平均内衬间隔和DI值均呈显著正相关(r值为0.694～0.754,均P＜0.05);S12及S24组小鼠CD8+IL-21R+细胞比例与平均内衬间隔和DI呈显著正相关(r值分别为0.516和0.725均P＜0.05).结论 香烟暴露导致肺气肿小鼠肺内Th17细胞数量及活性上调,并随烟雾暴露时间延长而增强;Th17细胞通过IL-21及IL-21R在肺部Th1/Tc1炎症中起重要促进作用;这对探讨COPD肺部炎症和肺气肿发生机制以及新的治疗靶点具有重要意义.
目的 觀察香煙煙霧暴露小鼠肺實質中CD4+白細胞介素(IL)-17+輔助性T細胞(Th17)數量及活性的錶達,探討其在香煙暴露小鼠肺部CD4+ γ-榦擾素+(Th1)炎癥及肺氣腫中的作用及相關機製.方法 將40隻雄性Balb/c小鼠按隨機數字錶法分為4組:對照12週(C12)組、對照24週(C24)組、煙霧暴露12週(S12)組、煙霧暴露24週(S24)組,每組10隻.香煙煙霧暴露法建立小鼠肺氣腫模型.HE染色觀察小鼠肺氣腫的改變,計算平均內襯間隔和肺泡破壞指數(DI);流式細胞術檢測小鼠肺實質中CD4+IL-17+T(Th17)細胞、CD4+γ-榦擾素+T(Th1)細胞、CD4+IL-17+γ-榦擾素+T(Th17/Th1)細胞、CD8+γ-榦擾素+T(Tc1)細胞、CD8+IL-21R+細胞及CD4+IL-17+IL-21+細胞比例;熒光定量PCR法檢測小鼠肺實質中維甲痠相關孤獨受體(RORγt)和IL-17的mRNA錶達,併分析這些指標的相互關繫.結果 S12組和S24組的平均內襯間隔[(39±4)μm和(47±7)μm]和DI(39.1±1.6和45.2±3.1)明顯高于C12組[(32±4)μm和28.2±1.6]和C24組[(33±3)μm和28.9±2.1],且以S24組的增高更為明顯,差異均有統計學意義(t值為4.378～15.188,均P＜0.05);S12組和S24組Th17細胞比例[(3.3±1.1)%和(7.2±2.2)%]均明顯高于C12組和C24組[(1.8±0.8)%和(2.0±0.6)%];S12組和S24組RORγtmRNA錶達量[(25±4)和(35±3)]及IL-17的mRNA錶達量[(26±3)和(36±3)]亦明顯高于C12組[(10±5)和(13±5)]和C24組[(11±7)和(8±6)],以S24組增高更為明顯,差異均有統計學意義(P＜0.05);S12組和S24組Th1細胞比例[(10.0±3.7)%和(26.2 ±6.0)%]、Th17/Th1細胞比例[(0.61±0.30)%和(1.82±0.52)%]及Tc1細胞比例[(17.0±4.5)%和(26.8±8.5)%]均明顯高于C12組[(3.8±1.7)%、(0.27±0.17)%和(4.8±1.9)%]和C24組[(4.2±1.3)%、(0.28±0.11)%和(5.2±1.0)%],以S24組增高更為明顯,差異均有統計學意義(P＜0.05);S12組和S24組小鼠Th17細胞與Th1、Tc1細胞比例、平均內襯間隔、DI值均呈顯著正相關(r值為0.519～0.797,均P＜0.01);Th17/Th1細胞比例與平均內襯間隔、DI值呈顯著正相關(r值分彆為0.742和0.802,均P＜0.01);S12組和S24組CD4+IL-17+IL-21+細胞比例[(0.19±0.04)和(0.55±0.24)]明顯高于C12組和C24組[(0.07±0.03)和(0.08±0.03)],S24組增高更為明顯,差異均有統計學意義(P＜0.05).S12組和S24組的CD8+IL-21R+細胞比例[(2.94±1.26)和(4.12±2.26)]高于C12組和C24組[(1.22±0.31)和(1.34±0.18)](P＞0.05);S12組及S24組小鼠CD4+IL-17+IL-21+細胞比例與Th1、Tc1細胞比例、平均內襯間隔和DI值均呈顯著正相關(r值為0.694～0.754,均P＜0.05);S12及S24組小鼠CD8+IL-21R+細胞比例與平均內襯間隔和DI呈顯著正相關(r值分彆為0.516和0.725均P＜0.05).結論 香煙暴露導緻肺氣腫小鼠肺內Th17細胞數量及活性上調,併隨煙霧暴露時間延長而增彊;Th17細胞通過IL-21及IL-21R在肺部Th1/Tc1炎癥中起重要促進作用;這對探討COPD肺部炎癥和肺氣腫髮生機製以及新的治療靶點具有重要意義.
목적 관찰향연연무폭로소서폐실질중CD4+백세포개소(IL)-17+보조성T세포(Th17)수량급활성적표체,탐토기재향연폭로소서폐부CD4+ γ-간우소+(Th1)염증급폐기종중적작용급상관궤제.방법 장40지웅성Balb/c소서안수궤수자표법분위4조:대조12주(C12)조、대조24주(C24)조、연무폭로12주(S12)조、연무폭로24주(S24)조,매조10지.향연연무폭로법건립소서폐기종모형.HE염색관찰소서폐기종적개변,계산평균내츤간격화폐포파배지수(DI);류식세포술검측소서폐실질중CD4+IL-17+T(Th17)세포、CD4+γ-간우소+T(Th1)세포、CD4+IL-17+γ-간우소+T(Th17/Th1)세포、CD8+γ-간우소+T(Tc1)세포、CD8+IL-21R+세포급CD4+IL-17+IL-21+세포비례;형광정량PCR법검측소서폐실질중유갑산상관고독수체(RORγt)화IL-17적mRNA표체,병분석저사지표적상호관계.결과 S12조화S24조적평균내츤간격[(39±4)μm화(47±7)μm]화DI(39.1±1.6화45.2±3.1)명현고우C12조[(32±4)μm화28.2±1.6]화C24조[(33±3)μm화28.9±2.1],차이S24조적증고경위명현,차이균유통계학의의(t치위4.378～15.188,균P＜0.05);S12조화S24조Th17세포비례[(3.3±1.1)%화(7.2±2.2)%]균명현고우C12조화C24조[(1.8±0.8)%화(2.0±0.6)%];S12조화S24조RORγtmRNA표체량[(25±4)화(35±3)]급IL-17적mRNA표체량[(26±3)화(36±3)]역명현고우C12조[(10±5)화(13±5)]화C24조[(11±7)화(8±6)],이S24조증고경위명현,차이균유통계학의의(P＜0.05);S12조화S24조Th1세포비례[(10.0±3.7)%화(26.2 ±6.0)%]、Th17/Th1세포비례[(0.61±0.30)%화(1.82±0.52)%]급Tc1세포비례[(17.0±4.5)%화(26.8±8.5)%]균명현고우C12조[(3.8±1.7)%、(0.27±0.17)%화(4.8±1.9)%]화C24조[(4.2±1.3)%、(0.28±0.11)%화(5.2±1.0)%],이S24조증고경위명현,차이균유통계학의의(P＜0.05);S12조화S24조소서Th17세포여Th1、Tc1세포비례、평균내츤간격、DI치균정현저정상관(r치위0.519～0.797,균P＜0.01);Th17/Th1세포비례여평균내츤간격、DI치정현저정상관(r치분별위0.742화0.802,균P＜0.01);S12조화S24조CD4+IL-17+IL-21+세포비례[(0.19±0.04)화(0.55±0.24)]명현고우C12조화C24조[(0.07±0.03)화(0.08±0.03)],S24조증고경위명현,차이균유통계학의의(P＜0.05).S12조화S24조적CD8+IL-21R+세포비례[(2.94±1.26)화(4.12±2.26)]고우C12조화C24조[(1.22±0.31)화(1.34±0.18)](P＞0.05);S12조급S24조소서CD4+IL-17+IL-21+세포비례여Th1、Tc1세포비례、평균내츤간격화DI치균정현저정상관(r치위0.694～0.754,균P＜0.05);S12급S24조소서CD8+IL-21R+세포비례여평균내츤간격화DI정현저정상관(r치분별위0.516화0.725균P＜0.05).결론 향연폭로도치폐기종소서폐내Th17세포수량급활성상조,병수연무폭로시간연장이증강;Th17세포통과IL-21급IL-21R재폐부Th1/Tc1염증중기중요촉진작용;저대탐토COPD폐부염증화폐기종발생궤제이급신적치료파점구유중요의의.
Objective To evaluate the expression and the role of Th 17 in cigarette smoke-induced lung inflammation and emphysema in mice.Methods Forty male BALB/c mice were randomly divided into 4 groups, including a control group C12, a control group C24, a smoke-exposure 12 week group (S12) and a smoke-exposure 24 week group S24 (n = 10 each).Morphological changes were evaluated by mean linear intercepts and destructive index (DI).The proportion of CD4+ IL-17 + Th17, CD4+ IFN-γ+ Th1, CD4+ IL-17 +IFN-γ+ T( Th17/ Th1 ), CD8+ IFN-γ+ Tc1, CD8+ IL-21R + and CD4+ IL-17 + IL-21 + T cells in lungs of mice was determined by flow cytometry.The mRNA expressions of RORγt and IL-17 were evaluated by real-time PCR.Results Mean linear intercepts and DI were significantly higher in S12 and S24 groups [(39 ± 4)μm, (47 ±7) μm], (39.1 ± 1.6, 45.2 ±3.1 ) as compared to C12[(32 ±4) μm,28.2 ± 1.6] and C24groups [(33 ± 3 ) μm ,28.9 ± 2.1], all P ＜ O.05.The percentage of Th17 of S12 and S24 groups [(3.3 ±1.1 )%, (7.2 ±2.2)%] was significantly increased as compared with that of C12 and C24 groups [( 1.8± 0.8) %, (2.0 ± 0.6) %], all P ＜ 0.05.The mRNA levels of RORγt [( 25 ± 4), ( 35 ± 3 )] and IL-17 [(26 ± 3), (36 ± 3 )] in S12 and S24 groups were higher than in C12 [(10 ± 5 ), (13 ± 5 )] and C24 groups [( 11 ± 7 ), (8 ± 6)], all P ＜ 0.05.The percentage of Th 1, Th17/Th1 and Tc1 cells of S12 and S24 groups [(10.0 ±3.7)%, (26.2 ±6.0)%], [(0.61 ±0.30)%, (1.82 ±0.52)%], [(17.0±4.5 ) %, ( 26.8 ± 8.5 ) %] was significantly increased as compared with that of C12 [( 3.8 ± 1.7 ) %,(0.27±0.17)%, (4.8 ±1.9)%] and C24 groups [(4.2±1.3)%, (0.28±0.11)%, (5.2±1.0)%], all P＜0.05.Moreover, the frequency of Th17 cells had a positive correlation with Th1, Tc1 cells and emphysematous lesions ( r =0.519 - 0.797, all P ＜ 0.01 ).In addition, a positive correlation between Th17/Th1 cells and emphysematous lesions was also found (r =0.742, 0.802, all P ＜0.01 ).The percentage of CD4+ IL-17+ IL-21 +T cells was significantly increased in S12 and S24 groups [(0.19 ±0.04) %, (0.55 ± 0.24) %] compared to controls [(0.07 ± 0.03 ) %, (0.08 ± 0.03 ) %], all P ＜ 0.05.Meanwhile, as compared with that of the controls [( 1.22 ± 0.31 ), ( 1.34 ± 0.18 )], the percentage of CD8+ IL-21 R + T cells was also increased in SI 2 and S24 groups [( 2.94 ± 1.26 ), (4.12 ± 2.26 )], but there were no differences among smoke-exposure groups ( P ＞0.05 ).The frequency of CD4+ IL-17 + IL-21 + T cells had a positive correlation with Th 1, Tc1 cells and emphysematous lesions (r = 0.694 -0.754, all P ＜0.05).And the frequency of CD8+ IL-21R+ T cells also had a positive correlation with emphysematous lesions ( r = 0.516, 0.725, all P ＜ 0.05).Conclusions Cigarette smoke increased the expression and the activity of Th17 in mice.Th17 may play a potential (active) role in the development of lung inflammation through IL-21/IL-21R pathway.
