目的探讨CD40对烟草烟雾暴露肺气肿小鼠肺部CD8^+T细胞毒性功能的影响。方法40只雄性C57小鼠按随机数字表法随机分为CD40^+/+对照组、CD40^+/+烟熏组、CD40^-/-对照组、CD40^-/-烟熏组4组,烟熏组采用烟草烟雾暴露24周建立小鼠肺气肿模型。各组肺组织分别采用HE染色观察肺部病理改变并计算肺泡平均内衬间隔;免疫组织化学法检测CD8^+T细胞和穿孔素^+、颗粒酶B^+细胞百分比;实时荧光定量PCR(RT—PCR)检测穿孔素、颗粒酶B、白细胞介素(IL)-27mRNA表达;酶联免疫吸附(ELISA)法检测IL-27细胞因子水平。结果CD40^+/+烟熏组肺组织明显破坏,可见大量炎症细胞浸润;CD40^-/-烟熏组肺组织破坏及炎症细胞浸润程度较CD40^+/+烟熏组轻。CD40^+/+烟熏组肺泡平均内衬间隔显著高于CD40^+/+对照组、CD40^-/-对照组和CD40^-/-烟熏组[(37.2±3.6)比(24.0±3.4)、(22.5±2.4)、(29.9±1.7)μm](均P〈0.05);CD40。一烟熏组高于CINO“’对照组及CD40。一对照组(均P〈0.05)。CD40“’烟熏组CD8^+、穿孔素^-、颗粒酶B^+细胞百分比[(16.3±2.3)%、(11.4±2.1)%、(10.7±1.9)%]均显著高于CD40^+/+对照组[(8.3±1.6)%、(5.1±1.2)%、(4.6±1.0)%]、CD40^-/-对照组[(6.4±1.5)%、(4.3±1.0)%、(4.2±1.0)%]、CI40^-/-烟熏组[(8.6±1.7)%、(5.6±1.3)%、(5.5±1.3)%](均P〈0.05)。CD40^+/+烟熏组穿孔素、颗粒酶B、IL-27mRNA相对表达量[(20.3士7.3)、(18.3±12.3)、(2.24-0.7)]均显著高于CIMO^+/+对照组[(9.4士4.8)、(10.6±3.8)、(1.3±0.6)]、CD40^-/-对照组[(8.1±3.1)、(7.7±3.5)、(1.1±0.5)]、CD40^-/-烟熏组[(12.94-6.2)、(10.4±4.6)、(1.5±0.4)](均P〈0.05)。CD^?
Objective To explore the effect of CIM0 knock out on the cytotoxic function of CD8 ~ T cell of mice with eigarette smoke-induced emphysema. Methods A total of 40 male C57 mice were divided into four groups according to the random number table, including CD40 ^+/+ control group, CD40^+/+ smoke- exposure group, CD40^-/-control group, CD40 ^-/-smoke-exposure group. The smoke-exposure groups were exposed to cigarette smoke for 24 weeks to establish emphysema model. Morphological changes were evaluated by linear intercepts. The percentages of CD8, perforin, granzyme B positive eells were evaluated by immunohistochemistry. The mRNA expressions of perforin, granzyme B, interleukin (IL) -27 were measured by fluorescent real time quantitative polymerase chain reaction (RT-PCR). The IL-27 cytokine level was tested by enzyme-linked immunosorbent assay (ELISA). Results The mean linear intercepts in CD40^+/+ smoke-exposure group was significantly higher than CD40^+/+ control group, CD40^-/- control group, and CD40^ -/- smoke-exposure group [ ( 37.2 ± 3.6 ) vs ( 24. 0 ± 3.4 ), ( 22. 5± 2.4 ), ( 29. 9 ±1.7) μm] ( all P 〈 0. 05 ) . CD40^-/- smoke-exposure group was higher than CD40^+/+ control group, CD40^ -/ -control group (all P 〈 0. 05 ). The percentages of CD8 positive, perforin positive and granzyme B positive cells in CD40 +/+ smoke-exposure group [ ( 16. 3 ± 2. 3 ) %, ( 11.4 ± 2. 1 ) %, ( 10. 7 ± 1.9 ) % ] were significantly higher than CD40^+/+ control group [ (8.3 ± 1.6)%, (5.1 ± 1.2)%, (4. 6 ± 1.0)% ], CD40^ -/- control group [ (6.4 ± 1. 5 ) % , (4. 3 ± 1.0) % , (4. 2 ± 1.0) % ] and CD40^ -/- smoke-exposure group [(8.6±1.7)%, (5.6±1.3)%, (5.5±1.3)%] (allP〈0.05). RT-PCR results showed that the mRNA expressions of perforin, granzyme B and IL-27 in CD40 ^+/+ smoke-exposure group [ (20. 3 ± 7.3), ( 18.3 ± 12. 3), (2.2 ±0. 7) ] were significantly higher than CD40 ^+/