Toll样受体2在七氟醚抑制哮喘小鼠气道炎症中的作用

Effect of Toll-like receptor 2 on the inhibition role of sevoflurane on airway inflammation in asthmatic mice

目的探讨Toll样受体2（TLR2）在七氟醚抑制哮喘小鼠气道炎症中的作用。方法C57 BL／6小鼠肺组织标本为前期研究所获取，分为C57对照组、C57哮喘组、C57七氟醚干预组各8份。24只SPF级雌性TLR2基因缺失（TLR2^-/-）小鼠按随机数字表法分为TLR2^-/-对照组、TLR2^-/-哮喘组、TLR2^-/-七氟醚干预组各8只。卵清蛋白致敏和激发建立哮喘模型，七氟醚干预组采用3％七氟醚反复吸入干预。免疫印迹检测C57各组小鼠肺组织TLR2表达情况；TLR2^-/-小鼠：分类计数肺泡灌洗液（BALF）炎症细胞，酶联免疫吸附（ELISA）法测定BALF中肿瘤坏死因子α（TNF-α）与白细胞介素10（IL-10）的表达；HE染色观察肺组织炎症情况。结果C57哮喘组肺组织TLR2相对表达量（0．547±0．042）显著高于C57对照组（0．312±0．023）（P=0．023）和C57七氟醚干预组（0．287±0．033）（P=0．020）；TLR2^-/-七氟醚干预组炎症细胞总数[（83．13±19．43）×10^3个／ml]、各种炎症细胞分类计数、TNF—α表达[（546±16）pg／m1]均显著低于TLR2^-/-哮喘组[（206．43±41．82）×10^3个／ml、（732±41）pg／m1]，但仍显著高于TLR2^-/-对照组[（44．64±7．17）×10^3个／ml、（380±24）pg／ml]（均P〈0．05）；TLR2^-/-七氟醚干预组肺组织炎症细胞浸润显著少于TLR2^-/-哮喘组。结论TLR2参与介导了七氟醚对哮喘小鼠气道炎症的抑制作用。

Objective To investigate the effect of Toll-like receptor 2 （TLR2） on the inhibition role of sevoflurane on airway inflammation in asthmatic mice. Methods The lung tissue samples of C57 BL/6 mice used in this study were from previous research, including control group, asthma group and sevoflurane group, 8 samples in each group. Twenty-four specific pathogen free female TLR2 gene deletion （TLR2^ -/- ） mice were randomly assigned to control group, asthma group and sevoflurane group, with 8 mice in each group. Asthma group and sevoflurane group were then sensitized and challenged with ovalbumin （OVA） to establish asthma model, combined with repeated inhalation of 3% sevoflurane in sevoflurane group. In C57 mice, expression levels of TLR2 were detected using Western blotting analyses. In TLR2 ^-/- mice, numbers of differential inflammatory cells were investigated; levels of tumor necrosis factor-alpha （TNF-ct） and interleukin-10 （IL-10） in bronchoalveolar lavage fluid （BALF） were measured by enzyme linked immunosorbent assay （ELISA） ; lung tissue inflammation was detected with HE staining. Results In lung tissues from C57 mice, levels of protein expression of TLR2 in asthma group （0. 547 ±0. 042） were higher than those in control group （0. 312 ±0. 023 ） （ P = 0. 023 ） and sevoflurane group （0. 287 ±0. 033 ） （ P = 0. 020）. In TLR2^-/- mice, the number of total cells （ （83.13 ± 19.43） ×10^3/ml）, numbers of differential inflammatory cells and TNF-α level （（546 ±16） pg/ml） in BALF in sevoflurane group were lower than those in asthma group （ （206.43 ±41.82） × 10^3/ml, （732± 41 ） pg/ml）, but still higher than those in control group （ （44. 64 ±7. 17 ） ×10^3/ml, （ 380 ±24 ） pg/ml ） （ all P 〈 0. 05 ） ; lung tissue inflammation was inhibited in sevoflurane group than in asthma group, but still more obvious than that in control group. Conclusion Toll like receptor 2 involved in the anti-inflammatory effect of sevoflurane on asthmatic airway inflammation in mice.