中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2011年
28期
1996-2000
,共5页
段敏超%钟小宁%黄颖%何志义%唐海娟
段敏超%鐘小寧%黃穎%何誌義%唐海娟
단민초%종소저%황영%하지의%당해연
肺气肿%T淋巴细胞,辅助诱导%受体,趋化因子%白细胞介素类%烟草对健康的危害
肺氣腫%T淋巴細胞,輔助誘導%受體,趨化因子%白細胞介素類%煙草對健康的危害
폐기종%T림파세포,보조유도%수체,추화인자%백세포개소류%연초대건강적위해
Pulmonary emphysema%T-lymphocytes,helper-inducer%Receptors,chemokine%Interleukins%Tobacco use disorder
目的 探讨香烟烟雾暴露肺气肿小鼠肺实质中Th17细胞的变化及其可能机制.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周组、对照24周组、烟雾暴露12周组、烟雾暴露24周组,每组10只.香烟烟雾暴露法建立小鼠肺气肿模型.HE染色观察小鼠肺组织的改变,计算平均内衬间隔和肺泡破坏指数;流式细胞术检测小鼠肺实质中Th17细胞、Th1细胞、Th17/Th1细胞、CD4+IL-17+趋化因子受体(CCR)6+T细胞占CD4细胞比例及CCR6+Th17细胞占Th17细胞比例;酶联免疫吸附法(ELISA)检测小鼠肺实质中的白细胞介素(IL)-1β、IL-6、IL-23、转化生长因子(TGF)-β、γ-干扰素(IFN-γ)及趋化因子(CCL)20水平,并分析其相互关系.结果 烟雾暴露12周组和24周组小鼠平均内衬间隔分别为(39±4)μm和(47±7)μm,肺泡破坏指数分别为(39.1±1.6)和(45.2±3.1),明显高于对照12周组[(33±3)μm和(28.2±1.6)]和24周组[(32±4)μm和(28.9±2.1)],以烟雾暴露24周组增高更为显著(均P<0.05);Th17细胞比例分别为(3.27±1.12)和(7.19±2.24)、Th17/Th1细胞比例分别为(0.61±0.30)和(1.82±0.52)、Th1细胞比例分别为(10.02±3.68)和(26.21±6.04)也均高于对照12周组[(1.80±0.75),(0.27±0.12)和(3.75±1.72)]和24周组[(1.99±0.59),(0.28±0.11)和(4.16±1.32)](均P＜0.01),且与平均内衬间隔、肺泡破坏指数呈正相关(r=0.706～0.877,均P＜0.01);CD4+IL-17+CCR6+T细胞比例分别为(0.69±0.34)和(1.11±0.48),CCR6+Th17细胞比例分别为(12.23±2.13)和(18.65±1.17),高于对照12周组[(0.22±0.18)和(6.55±2.13)]和24周组[(0.25±0.17)和(7.29±1.57)],以烟雾暴露24周组增高更为显著(均P＜0.05),其中CCR6+Th17细胞比例与平均内衬间隔和肺泡破坏指数呈正相关(r=0.617、0.741,均P＜0.05);IFN-γ、IL-1β、IL-6、IL-23、TGF-β及CCL20浓度也高于对照12周组和24周组(均P＜0.01),且与Th17细胞比例呈正相关(r=0.394～0.800,均P<0.05).结论 香烟烟雾暴露肺气肿小鼠肺内Th17细胞增高;这可能与IFN-γ、IL-1β、IL-6、IL-23、TGF-β的升高及CCR6/CCL20趋化轴作用有关.
目的 探討香煙煙霧暴露肺氣腫小鼠肺實質中Th17細胞的變化及其可能機製.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週組、對照24週組、煙霧暴露12週組、煙霧暴露24週組,每組10隻.香煙煙霧暴露法建立小鼠肺氣腫模型.HE染色觀察小鼠肺組織的改變,計算平均內襯間隔和肺泡破壞指數;流式細胞術檢測小鼠肺實質中Th17細胞、Th1細胞、Th17/Th1細胞、CD4+IL-17+趨化因子受體(CCR)6+T細胞佔CD4細胞比例及CCR6+Th17細胞佔Th17細胞比例;酶聯免疫吸附法(ELISA)檢測小鼠肺實質中的白細胞介素(IL)-1β、IL-6、IL-23、轉化生長因子(TGF)-β、γ-榦擾素(IFN-γ)及趨化因子(CCL)20水平,併分析其相互關繫.結果 煙霧暴露12週組和24週組小鼠平均內襯間隔分彆為(39±4)μm和(47±7)μm,肺泡破壞指數分彆為(39.1±1.6)和(45.2±3.1),明顯高于對照12週組[(33±3)μm和(28.2±1.6)]和24週組[(32±4)μm和(28.9±2.1)],以煙霧暴露24週組增高更為顯著(均P<0.05);Th17細胞比例分彆為(3.27±1.12)和(7.19±2.24)、Th17/Th1細胞比例分彆為(0.61±0.30)和(1.82±0.52)、Th1細胞比例分彆為(10.02±3.68)和(26.21±6.04)也均高于對照12週組[(1.80±0.75),(0.27±0.12)和(3.75±1.72)]和24週組[(1.99±0.59),(0.28±0.11)和(4.16±1.32)](均P＜0.01),且與平均內襯間隔、肺泡破壞指數呈正相關(r=0.706～0.877,均P＜0.01);CD4+IL-17+CCR6+T細胞比例分彆為(0.69±0.34)和(1.11±0.48),CCR6+Th17細胞比例分彆為(12.23±2.13)和(18.65±1.17),高于對照12週組[(0.22±0.18)和(6.55±2.13)]和24週組[(0.25±0.17)和(7.29±1.57)],以煙霧暴露24週組增高更為顯著(均P＜0.05),其中CCR6+Th17細胞比例與平均內襯間隔和肺泡破壞指數呈正相關(r=0.617、0.741,均P＜0.05);IFN-γ、IL-1β、IL-6、IL-23、TGF-β及CCL20濃度也高于對照12週組和24週組(均P＜0.01),且與Th17細胞比例呈正相關(r=0.394～0.800,均P<0.05).結論 香煙煙霧暴露肺氣腫小鼠肺內Th17細胞增高;這可能與IFN-γ、IL-1β、IL-6、IL-23、TGF-β的升高及CCR6/CCL20趨化軸作用有關.
목적 탐토향연연무폭로폐기종소서폐실질중Th17세포적변화급기가능궤제.방법 장40지웅성Balb/c소서안수궤수자표법분위4조:대조12주조、대조24주조、연무폭로12주조、연무폭로24주조,매조10지.향연연무폭로법건립소서폐기종모형.HE염색관찰소서폐조직적개변,계산평균내츤간격화폐포파배지수;류식세포술검측소서폐실질중Th17세포、Th1세포、Th17/Th1세포、CD4+IL-17+추화인자수체(CCR)6+T세포점CD4세포비례급CCR6+Th17세포점Th17세포비례;매련면역흡부법(ELISA)검측소서폐실질중적백세포개소(IL)-1β、IL-6、IL-23、전화생장인자(TGF)-β、γ-간우소(IFN-γ)급추화인자(CCL)20수평,병분석기상호관계.결과 연무폭로12주조화24주조소서평균내츤간격분별위(39±4)μm화(47±7)μm,폐포파배지수분별위(39.1±1.6)화(45.2±3.1),명현고우대조12주조[(33±3)μm화(28.2±1.6)]화24주조[(32±4)μm화(28.9±2.1)],이연무폭로24주조증고경위현저(균P<0.05);Th17세포비례분별위(3.27±1.12)화(7.19±2.24)、Th17/Th1세포비례분별위(0.61±0.30)화(1.82±0.52)、Th1세포비례분별위(10.02±3.68)화(26.21±6.04)야균고우대조12주조[(1.80±0.75),(0.27±0.12)화(3.75±1.72)]화24주조[(1.99±0.59),(0.28±0.11)화(4.16±1.32)](균P＜0.01),차여평균내츤간격、폐포파배지수정정상관(r=0.706～0.877,균P＜0.01);CD4+IL-17+CCR6+T세포비례분별위(0.69±0.34)화(1.11±0.48),CCR6+Th17세포비례분별위(12.23±2.13)화(18.65±1.17),고우대조12주조[(0.22±0.18)화(6.55±2.13)]화24주조[(0.25±0.17)화(7.29±1.57)],이연무폭로24주조증고경위현저(균P＜0.05),기중CCR6+Th17세포비례여평균내츤간격화폐포파배지수정정상관(r=0.617、0.741,균P＜0.05);IFN-γ、IL-1β、IL-6、IL-23、TGF-β급CCL20농도야고우대조12주조화24주조(균P＜0.01),차여Th17세포비례정정상관(r=0.394～0.800,균P<0.05).결론 향연연무폭로폐기종소서폐내Th17세포증고;저가능여IFN-γ、IL-1β、IL-6、IL-23、TGF-β적승고급CCR6/CCL20추화축작용유관.
Objective To evaluate the expression of Th17 cell in a cigarette smoke-induced mice model of emphysema and explore the probable mechanisms of its elevation. Methods Forty male Balb/c mice were randomly divided into 4 groups: control group for 12 weeks (C12), control group for 24 weeks (C24), smoke-exposure group for 12 weeks (S12) and smoke-exposure group for 24 weeks (S24)（n=10 each）. Morphological changes were evaluated by mean linear intercepts (Lm) and destructive index (DI). The percentages of Th17, Th1, Th17/Th1, CD4+IL-17+CCR6+T and CCR6+Th17 cells were determined by tetra-color flow cytometry while the levels of interleukin （IL）-1β, IL-6, IL-23, transforming growth factor (TGF)-β, interferon (IFN)-γ and CC chemokine ligand (CCL)-20 assayed by enzyme-linked immunosorbent assay (ELISA). Results The values of Lm [(39±4) μm, (47±7) μm] and DI [(39.1±1.6), (45.2±3.1)] were significantly higher in S12 and S24 than those in C12 [(33±3) μm, (28.2±1.6)] and C24 [(32±4) μm, (28.9±2.1)], particularly in C24 (all P<0.05). The percentages of Th17 cell [(3.27±1.12), (7.19±2.24)], Th17/Th1 cell [(0.61±0.30), (1.82±0.52)] and Th1 cell [(10.02±3.68), (26.21±6.04)] in the lungs of S12 and S24 significantly increased than those in C12 [(1.80±0.75), (0.27±0.12), (3.75±1.72)] and C24 [(1.99±0.59), (0.28±0.11), (4.16±1.32)], particularly in C24 (all P<0.01). The percentages of Th17, Th17/Th1 and Th1 cells in the lungs of S12 and S24 had a positive correlation with Lm and DI (all P<0.01). The percentages of CD4+IL-17+CCR-6+T cell [(0.69±0.34), (1.11±0.48)] and CCR6+Th17 cell [(12.23±2.13), (18.65±1.17)] were significantly elevated in S12 and S24 compared to those in C12 [(0.22±0.18), (6.55±2.13)] and C24 [(0.25±0.17), (7.29±1.57)], particularly in C24 (all P<0.05). Furthermore, a positive correlation between CCR6+Th17 cell and emphysematous lesions was also found (all P<0.05). The levels of IL-1β, IL-6, IL-23, TGF-β, IFN-γ and CCL20 significantly increased in S12 and S24 as compared with those of C12 and C24 (all P<0.05). Meanwhile, the percentage of Th17 cell had a positive correlation with IL-1β, IL-6, IL-23, TGF-β, IFN-γ and CCL20. Conclusion There is an up-regulated expression of Th17 in lungs of cigarette smoke-induced emphysema mice. The CCR6/CCL20 axis and the elevated levels of IL-1β, IL-6, IL-23, TGF-β and IFN-γ may be related with the above effect.
