肝郁对哮喘大鼠辅助性T细胞17和调节性T细胞的影响

Effect of liver depression on Thl7 and treg in asthma ratsi

目的研究肝郁（肝气郁结证）对哮喘大鼠辅助性T淋巴细胞17（Th17）、调节性T细胞（Treg）、白细胞介素．17（IL-17）、白细胞介素．10（IL-10）及气道炎症反应的影响，阐明肝郁型哮喘发病的免疫机制。方法利用中医肝郁证动物模型和西医哮喘动物模型的建模方法，建立大鼠肝郁型哮喘的病证结合模型。收集支气管肺泡灌洗液（BALF）并进行细胞总数及分类计数、肺组织进行病理学观察；流式细胞术检测外周血中Thl7和Treg细胞占CD4十T细胞比例；酶联免疫吸附试验（ELISA）法检测大鼠血清中IL-17、IL-10的水平。结果肝郁哮喘组、哮喘模型组炎性细胞总数为[（96．86±4．43）×10^7／L、（88．22±3．22）×10^7／L]、嗜酸性粒细胞比例为[（27．58±4．65）％、（22．67±2．43）％]、Thl7细胞比例为[（6．86±0．98）％、（6．01±0．77）％]、IL-17水平为[（48．88±8．06）pg／ml、（43．24±6．32）pg／m1]均高于正常对照组[（30．58±2．49）x10^7／L、（O．78±0．12）％、（2．80±0．82）％、（24．11±3．40）pg／m1]；肝郁哮喘组和哮喘模型组Treg细胞比例[（3．09±0．55）％、（3．96±0．66）％]及IL-10水平[（19．79±2．80）pg／ml、（20．29±3．12）pg／m1]均低于正常对照组[（8．02±1．26）％、（30．79±4．01）pg／m1]。肝郁哮喘组炎性细胞总数（96．86±4．43）×10^7／L、嗜酸性粒细胞比例（27．58±4．65）％、Thl7细胞比例（6．86±0．98）％、IL-17水平（48．88±8．06）pg／ml均高于哮喘模型组[（88．22±3．22）×10^7／L、（22．67±2．43）％、（6．01±0．77）％、（43．24±6．32）pedml]；肝郁哮喘组Treg细胞比例（3．09±0．55）％低于哮喘模型组（3．96±0．66）％；肝郁哮喘组肺组织病理学观察症状较哮喘模型组严重。结论肝气郁结证可促进哮喘的炎症反应、Thl7／Treg平衡失调和IL-17水平增加而加重哮喘。合并肝郁证是哮喘反复发作的重要内在因素之一。

[Abstract] Objective To investigate the effect of liver depression （Liver Qi Stagnation） on th17, Treg, IL-17, IL-10 and airway inflammation in asthmatic rats, and to clarify the immune mechanism of asthma with liver depression. Methods Established the combined with disease and syndrome model of asthma with liver depression. Collected the bronchoalveolar lavage fluid （BALF） to count the total and differential cell. Lung tissue was observed in microscope; the proportion of Thl7 cells and Trcg cells of CD4 tT cells in peripheral blood was measured by flow cytometry; the levels of IL-17 and IL-10 were determined by ELISA. Results The total number of inflammatory cells[（96.86±4.43）X 107/L, （88.22±3.22）Xl0V/L], the proportion of eosinophils [ （27.58±4.65） %, （22.67±2.43） %], Thl7 cells [ （6.86±0.98） %, （6.01 ±0.77） %] and IL-17 level [ （48.88± 8.06）pg/ml, （43.24± 6.32） pg/mlJ of asthma in liver depression group and asthma group were significantly higher than the control group [ （30.58±2.49） 10T/L, （0.78±0.12） %, （2.80±0.82） %, （24.11 ± 3.40） pg/ml] ; Treg ceils [ （3.09± 0.55） %, （3.96± 0.66） %] and IL-10 level [ （19.79± 2.80） pg/ml, （20.29± 3.12） pg/ml] were significantly lower than the control group [ （8.02± 1.26）%, （30.79± 4.01）pg/ml]. The total number of inflammatory ceils （96.86±4.43） 10S/L, the proportion of eosinophils （27.58± 4.65）% and Thl7 cells（6.86± 0.98） % and IL-17 level（48.88± 8.06）pg/mL of asthma in liver depression group were significantly higher than the asthma group （88.22± 3.22）X 10Y/L, （22.67± 2.43）%, （6.01± 0.77）%, （43.24± 6.32） pg/ml; the proportion of Treg cells （3.09± 0.55）% was significantly lower than the asthma group （3.96± 0.66）%; and the lung histopathology symptoms was more severe than asthma group. Coneluslon Liver Qi Stagnation can promote the inflammation of asthma, the imbalance of Th 17/Treg and IL-17 level to aggravate the asthma. Liver depression is one of the major internal factors in recurrent episodes of asthma.