CD40对烟草烟雾暴露小鼠肺组织Foxp3＋Treg细胞的影响  

Effect of CD40 on Foxp3＋ Treg cell in the lung of cigarette smoke exposure mice

作　　者：邓婷婷[1] 钟小宁[1] 王琴[1] 邝良鉴[1] 邱诗林[1] 梁毅[1] 何志义[1] 张建全[1] 白晶[1] 

机构地区：[1]广西医科大学第一附属医院呼吸内科,南宁530021

出　　处：《中华医学杂志》2016年第14期1139-1143,共5页National Medical Journal of China

基　　金：国家自然科学基金（81260013）

摘　　要：目的观察烟草烟雾暴露下CD40对小鼠肺内叉头／翼状螺旋转录因子阳性调节性T细胞（Foxp3＋Treg）的影响。方法野生型与CD40-/-C57BL／6小鼠各20只，分别按随机数字表法分为两组，即：野生型对照组、野生型烟熏组与CD40-/-对照组、CD40-/-烟熏组，每组各10只。烟熏组香烟烟雾暴露24周建立小鼠肺气肿模型，HE染色观察小鼠肺组织的改变，计算肺泡平均内衬间隔（MLI）。免疫组化法观察肺组织中Foxp3＋细胞百分数，实时定量PCR与Western印迹法检测肺组织中Foxp3mRNA与蛋白的表达。酶联免疫吸附（ELISA）法检测各组小鼠肺组织中白细胞介素（IL）-10，IL-35水平。结果CD40-/-烟熏组MLI显著低于野生型烟熏组[（30．0±1．7）比（37．3±3．7）μm]，高于CD40-/-对照组[（23．2±2．5）±m]，野生型烟熏组显著高于野生型对照组[（22．2±1．7）±m]（均P〈0．05）。CD40-/-烟熏组肺组织中Foxp3＋细胞数量均显著高于野生型烟熏组和CD40-/-对照组[（16．89±0．75）％比（9．65±0．74）％和（13．58±0．51）％]，野生型烟熏组显著低于野生型对照组[（12．13±0．81）％]（均P〈0．05）。CD40-/-烟熏组肺组织中Foxp3的mRNA与蛋白表达量均显著高于野生型烟熏组和CD40-/-对照组，野生型烟熏组显著低于野生型对照组（均P〈0．05）。CD40-/-烟熏组肺组织匀浆中IL-10水平均显著高于野生型烟熏组和CD40-/-对照组[（231±25）比（80±31）和（183±29）ng／L]，野生型烟熏组显著低于野生型对照组[（192±37）ng／L]（均P〈0．05）。CD40-/-烟熏组肺组织匀浆中IL．35水平均显著高于CD40。一对照组、野生型对照组、野生型烟熏组[（2084-29）比（118±29）、（148±36）、（137±37）ng／L]（均P〈0．05）。结论敲除CD40基因可促进烟草烟雾暴露小鼠肺内Treg细胞分化，阻断CD40-/-CD40配体通路可能有助于改善烟草烟雾暴露所致的�Objective To evaluate the effect of CD40 on Foxp3＋ Treg cell in the lung of cigarette smoke exposure mice. Methods According to the random number table, 20 wild type （WT） C57 BL/6 mice and 20 CD40-/-C57 BL/6 mice were randomly divided into two groups： WT control group, WT smoke-exposure group （24 weeks ） and CD40-/- control group, CD40-/- smoke-exposure group （24 weeks） （n = 10 each）. Alveolar airspace enlargement was observed by HE staining. Morphological change was evaluated by mean linear intercepts （MLI）. Immunohistoehemical method was used to detect the quantity of Foxp3 ＋ cell in the lung. The mRNA expression of Foxp3 was measured by fluorescence quantitative real-time polymerase chain reaction （qRT-PCR）. The protein level of Foxp3 was measured by Western blot. Interleukin （IL） -10 and IL-35 levels in the lung were tested by enzyme-linked immunosorbent assay （ELISA）. Results The MLI in CD40-/- smoke-exposure group was significantly lower than the WT smoke-exposure group[ （30.0 ± 1.7） vs （37.3±3.7）μm], but higher than the CD40-/- control group [ （23. 2 ±2. 5） μm], WT smoke-exposure group was significantly higher than the WT control group[ （22. 2 ± 1.7） μm ] （ all P 〈 0. 05 ）. The percentage of Foxp3＋ cell in the lungs of CD40 -/- smoke-exposure group was significantly higher than the WT smoke-exposure group and CD40-/- control group[ （ 16. 89 ± 0. 75）% vs （9. 65 ±0.74 ） % and （ 13.58 ±0. 51 ） % ], WT smoke-exposure group was significantly lower than WT control group[ （12. 13± 0. 81 ）% ] （all P 〈 0.05 ）. In the lungs, Foxp3 mRNA and protein expression in CD40 -/- smoke-exposure group were increased compared to WT-smoke-exposure group and CD40-/- control group, WT smoke-exposure group were decreased compared to WT control group （ all P 〈 0. 05 ）. In the lungs, the level of IL-10 in CD40-/- smoke-exposure group was higher than the WT smoke-exposure group and CD40-/- control group[ （231 ±25） vs （80 ±31�

关 键 词：TNF受体相关因子3 T淋巴细胞 调节性 肺气肿 小鼠 

分 类 号：R114[医药卫生—卫生毒理学]