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
2010年
9期
688-692
,共5页
邱诗林%白晶%钟小宁%黄秋嫔%陈慧%柳广南
邱詩林%白晶%鐘小寧%黃鞦嬪%陳慧%柳廣南
구시림%백정%종소저%황추빈%진혜%류엄남
T淋巴细胞,调节性%烟%戒烟%肺气肿
T淋巴細胞,調節性%煙%戒煙%肺氣腫
T림파세포,조절성%연%계연%폐기종
T-Lymphocytes,regulatory%Smoke%Smoking cessation%Pulmonary emphysema
目的 观察香烟暴露及断烟后大鼠Treg细胞的变化,探讨Treg细胞与断烟后大鼠肺部炎症反应和肺气肿持续存在的关系.方法 将50只雄性Wistar大鼠按随机数字表法分为5组:健康对照组、香烟暴露组和断烟组,其中健康对照组和香烟暴露组又分为12周组(对照1组和实验1组)和24周组(对照2组和实验2组).香烟烟雾暴露法建立大鼠肺气肿模型.HE染色观察大鼠肺气肿的改变,酶联免疫吸附法检测大鼠BALF中白细胞介素(IL-8)和肿瘤坏死因子-α(TNF-α)水平,流式细胞术检测大鼠外周血和肺实质中CD4+Foxp3调节性T细胞(Treg细胞)比例,实时定量PCR法检测大鼠外周血和肺实质中Foxp3的mRNA表达.多组间比较采用单因素方差分析,组内两两比较采用SNK和Games-Howell检验.结果 实验1组和实验2组平均内衬间隔(MLI)为(64.9±5.3)μm和(77.9±11.5)μm,均明显高于对照1组和对照2组的(39.0±3.8)μm和(40.3±2.7)μm,差异均有统计学意义(P＜0.01);断烟组MLI[(71.5 ±5.8)μm]介于实验1组和实验2组之间,但肺气肿程度较实验1组明显加重,差异有统计学意义(P＜0.01).实验1组和实验2组IL-8水平[(68±17)ng/L和(85±16)ng/L]及TNF-α水平[(14.1±1.8)ng/L和(20.1±8.7)ng/L]均明显高于对照1组和对照2组[(44±8)ng/L、(43±9)ng/L及(6.3±2.3)ng/L、(5.8±1.6)ng/L],差异均有统计学意义(P＜0.05).实验1组和实验2组肺实质中CD+4Foxp3+Treg细胞[(6.6±0.8)%和(5.3±0.9)%]明显少于对照1组和对照2组[(9.0±1.0)%和(9.6±0.9)%],差异均有统计学意义(P＜0.01);断烟组肺实质中CD+4Foxp3+Treg细胞[(7.2±0.6)%]与实验1组比较虽无明显减少,但未能恢复至正常水平.实验1组和实验2组肺实质中Foxp3的mRNA表达量(17±7和9±7)明显低于对照1组和对照2组(39±6和42±7),差异均有统计学意义(P＜0.01);断烟组肺实质中Foxp3的mRNA表达量(21±9)与实验1组比较未见明显减少,但未能恢复至正常水平.结论 香烟暴露肺气肿大鼠肺内调节性T细胞下调可能导致大鼠肺内炎症反应放大,并在断烟后持续存在,提示调节性T细胞下调可能是大鼠肺气肿持续进展的原因之一.
目的 觀察香煙暴露及斷煙後大鼠Treg細胞的變化,探討Treg細胞與斷煙後大鼠肺部炎癥反應和肺氣腫持續存在的關繫.方法 將50隻雄性Wistar大鼠按隨機數字錶法分為5組:健康對照組、香煙暴露組和斷煙組,其中健康對照組和香煙暴露組又分為12週組(對照1組和實驗1組)和24週組(對照2組和實驗2組).香煙煙霧暴露法建立大鼠肺氣腫模型.HE染色觀察大鼠肺氣腫的改變,酶聯免疫吸附法檢測大鼠BALF中白細胞介素(IL-8)和腫瘤壞死因子-α(TNF-α)水平,流式細胞術檢測大鼠外週血和肺實質中CD4+Foxp3調節性T細胞(Treg細胞)比例,實時定量PCR法檢測大鼠外週血和肺實質中Foxp3的mRNA錶達.多組間比較採用單因素方差分析,組內兩兩比較採用SNK和Games-Howell檢驗.結果 實驗1組和實驗2組平均內襯間隔(MLI)為(64.9±5.3)μm和(77.9±11.5)μm,均明顯高于對照1組和對照2組的(39.0±3.8)μm和(40.3±2.7)μm,差異均有統計學意義(P＜0.01);斷煙組MLI[(71.5 ±5.8)μm]介于實驗1組和實驗2組之間,但肺氣腫程度較實驗1組明顯加重,差異有統計學意義(P＜0.01).實驗1組和實驗2組IL-8水平[(68±17)ng/L和(85±16)ng/L]及TNF-α水平[(14.1±1.8)ng/L和(20.1±8.7)ng/L]均明顯高于對照1組和對照2組[(44±8)ng/L、(43±9)ng/L及(6.3±2.3)ng/L、(5.8±1.6)ng/L],差異均有統計學意義(P＜0.05).實驗1組和實驗2組肺實質中CD+4Foxp3+Treg細胞[(6.6±0.8)%和(5.3±0.9)%]明顯少于對照1組和對照2組[(9.0±1.0)%和(9.6±0.9)%],差異均有統計學意義(P＜0.01);斷煙組肺實質中CD+4Foxp3+Treg細胞[(7.2±0.6)%]與實驗1組比較雖無明顯減少,但未能恢複至正常水平.實驗1組和實驗2組肺實質中Foxp3的mRNA錶達量(17±7和9±7)明顯低于對照1組和對照2組(39±6和42±7),差異均有統計學意義(P＜0.01);斷煙組肺實質中Foxp3的mRNA錶達量(21±9)與實驗1組比較未見明顯減少,但未能恢複至正常水平.結論 香煙暴露肺氣腫大鼠肺內調節性T細胞下調可能導緻大鼠肺內炎癥反應放大,併在斷煙後持續存在,提示調節性T細胞下調可能是大鼠肺氣腫持續進展的原因之一.
목적 관찰향연폭로급단연후대서Treg세포적변화,탐토Treg세포여단연후대서폐부염증반응화폐기종지속존재적관계.방법 장50지웅성Wistar대서안수궤수자표법분위5조:건강대조조、향연폭로조화단연조,기중건강대조조화향연폭로조우분위12주조(대조1조화실험1조)화24주조(대조2조화실험2조).향연연무폭로법건립대서폐기종모형.HE염색관찰대서폐기종적개변,매련면역흡부법검측대서BALF중백세포개소(IL-8)화종류배사인자-α(TNF-α)수평,류식세포술검측대서외주혈화폐실질중CD4+Foxp3조절성T세포(Treg세포)비례,실시정량PCR법검측대서외주혈화폐실질중Foxp3적mRNA표체.다조간비교채용단인소방차분석,조내량량비교채용SNK화Games-Howell검험.결과 실험1조화실험2조평균내츤간격(MLI)위(64.9±5.3)μm화(77.9±11.5)μm,균명현고우대조1조화대조2조적(39.0±3.8)μm화(40.3±2.7)μm,차이균유통계학의의(P＜0.01);단연조MLI[(71.5 ±5.8)μm]개우실험1조화실험2조지간,단폐기종정도교실험1조명현가중,차이유통계학의의(P＜0.01).실험1조화실험2조IL-8수평[(68±17)ng/L화(85±16)ng/L]급TNF-α수평[(14.1±1.8)ng/L화(20.1±8.7)ng/L]균명현고우대조1조화대조2조[(44±8)ng/L、(43±9)ng/L급(6.3±2.3)ng/L、(5.8±1.6)ng/L],차이균유통계학의의(P＜0.05).실험1조화실험2조폐실질중CD+4Foxp3+Treg세포[(6.6±0.8)%화(5.3±0.9)%]명현소우대조1조화대조2조[(9.0±1.0)%화(9.6±0.9)%],차이균유통계학의의(P＜0.01);단연조폐실질중CD+4Foxp3+Treg세포[(7.2±0.6)%]여실험1조비교수무명현감소,단미능회복지정상수평.실험1조화실험2조폐실질중Foxp3적mRNA표체량(17±7화9±7)명현저우대조1조화대조2조(39±6화42±7),차이균유통계학의의(P＜0.01);단연조폐실질중Foxp3적mRNA표체량(21±9)여실험1조비교미견명현감소,단미능회복지정상수평.결론 향연폭로폐기종대서폐내조절성T세포하조가능도치대서폐내염증반응방대,병재단연후지속존재,제시조절성T세포하조가능시대서폐기종지속진전적원인지일.
Objective To evaluate the expression of Treg in a cigarette smoke-induced rat model of emphysema and after smoking cessation in the rats. Methods Fifty male Wistar rats were randomly divided into control group 1 (12 weeks), control group 2 (24 weeks), smoke-exposure group 1 (12 weeks),smoke-exposure group 2 (24 weeks) and smoking cessation group, with 10 rats in each group. Alveolar airspace enlargement was observed by hematoxylin-eosin (HE) staining. IL-8 and TNF-α levels in bronchoalveolar lavage fluid (BALF) were tested by ELISA. The proportion of CD4+ Foxp3+ Treg in peripheral blood and lungs of rats was determined by flow cytometry. The mRNA expression of Foxp3 was measured by real-time PCR. Comparisons of the data between different groups were performed using one-way ANOVA, and SNK and Games-Howell test was used for comparison between 2 groups. Results The mean linear intercept (MLI) in smoke-exposure group 1 and group 2 [( 64. 9 ± 5. 3 ) μm, (77.9 ± 11.5 ) μm] was higher than those in the control group 1 and group 2 [( 39. 0 ± 3. 8 ) μm, (40. 3 ± 2. 7 ) μm], all P ＜ 0. 01.Compared with smoke-exposure group 2, the MLI in smoking cessation group (71.5 ± 5. 8) μm showed a lower value ( P ＜ 0. 01 ), but still higher than that in smoke-exposure group 1 ( P ＜ 0. 01 ). The IL-8 and TNF-α levels in BALF of smoke-exposure group 1 and group 2 [(68 ± 17) ng/L, (85 ± 16) ng/L],[( 14. 1 ± 1.8)ng/L, (20. 1 ±8. 7) ng/L] were higher than those in control group 1 and group 2 [(44 ±8)ng/L, (43±9)ng/L], [(6.3 ±2.3) ng/L, (5.8±1.6)ng/L], all P＜0.05. The IL-8 and TNF-αlevels were not statistically different between in smoking cessation group (56±6)ng/L, (14.7±4.7)ng/L and smoke-exposure group 1. The percentage of Treg in the lungs of smoke-exposure group 1 and group 2 [(6. 6 ±0. 8)%, (5.3 ±0. 9)%] was significantly decreased as compared to control group 1 and group 2 [(9.0 ± 1.0 ) %, ( 9. 6 ± 0. 9 ) %], all P ＜ 0. 01. The percentage of Treg in lungs was not statistically different between smoke-exposure group 1 and smoking cessation group (7. 2 ±0. 6)%. In peripheral blood,there was no significant difference between groups in the percentage of Treg. In the lung, Foxp3 mRNA expression in smoke-exposure group 1 and group 2 [( 17 ±7), (9 ±7)] was less than that in control group 1 and group 2 [( 39 ± 6 ), (42 ± 7 )], all P ＜ 0. 01. The Foxp3 mRNA expression was not statistically different between smoke-exposure group 1 and smoking cessation group (21 ±9). No significant differences in peripheral blood Foxp3 mRNA expression was found between groups. Conclusions Decreased Treg was present in lungs of cigarette smoke-induced model of emphysema despite 12 weeks' smoking cessation,suggesting that down-regulation of Treg may be involved in the amplified and persistent inflammation after smoking cessation.
