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 o