【作者】 李进华；

【导师】 陈平； 陈燕；

【作者基本信息】 中南大学 ， 临床医学， 2014， 博士

【摘要】 目的：探讨腺病毒感染对CSE(cigarette smoke extract,香烟烟雾提取物)腹腔注射肺气肿小鼠肺功能、肺病理、肺部炎症、蛋白酶、凋亡及增殖的影响。方法：36只SPF级BALB/c小鼠随机分为PBS组、CSE组和CSE+Adenovirus组,每组12只,制备CSE。各组分别在第1、11、22天注射PBS或CSE,第15天气管内滴注PBS或腺病毒,第28天处理各组小鼠。病毒处理后第7天(造模第22天)每组随机选取2只小鼠制备肺组织冰冻切片观察RFP荧光；小动物肺功能检测系统检测肺功能；HE染色观察肺部病理形态学改变,测定平均内衬间隔(MLI)、肺泡破坏指数(DI)和平均肺泡隔厚度(MAST),行气道和血管炎症半定量评分；免疫组化法检测肺组织MMP-2和MMP-9表达；TUNEL法检测肺组织凋亡并计算凋亡指数(AI)；免疫组化法检测增殖标志物Ki67表达,反映肺组织增殖情况,并计算增殖指数(PI)。结果：(1) CSE+Adenovirus组肺组织可见红色荧光,PBS组和CSE组均未观察到红色荧光。(2)肺功能：与PBS组比较,CSE组与CSE+Adenovirus组小鼠肺阻力(RAW)增高,动态肺顺应性(Cdyn)降低,差异均具有统计学意义(p<0.05)。与CSE组比较,C SE+Adenovirus组RAW及Cdyn无明显差异(p>0.05)。各组间小鼠呼吸频率、潮气量、每分钟通气量、呼气时间、吸呼比差异均无统计学意义(p>0.05)。(3)肺病理：与PBS组比较,CSE组及CSE+Adenovirus组肺泡破坏更重,MLI和DI增高,差异有统计学意义(p<0.05),MAST降低,差异亦具有统计学意义(p<0.05)。与CSE组比较,CSE+Adenovirus组肺泡破坏程度无明显区别,MLI、DI及MAST均无明显区别,差异无统计学意义(p>0.05)。(4)气道及血管炎症：CSE+Adenovirus组气道管壁周围可见大量炎症细胞浸润,气道炎症半定量评分示,与PBS组比较,CSE组评分均值稍高于PBS组,但无统计学意义(p>0.05),CSE+Adenovirus组评分显著增高,有统计学意义(p<0.05)；与CSE组比较,CSE+Adenovirus组气道炎症评分也显著增高,有统计学意义(p<0.05)。CSE+Adenovirus组血管管壁周围可见炎症细胞浸润,血管炎症半定量分析显示,与PBS组比较,CSE组血管炎症评分无统计学差异(p>0.05), CSE+Adenovirus组血管炎症评分显著增高,有统计学意义(p0.05)；与CSE组比较,CSE+Adenovirus组血管炎症评分显著增高,有统计学意义(p0.05)。(5)MMP-2及MMP-9表达：与PBS组比较,CSE组和CSE+Adenovirus组MMP-2和MMP-9表达均增高,有统计学意义(p<0.05)；与CSE组比较,CSE+Adenovirus组该两组蛋白表达亦增高,有统计学意义(p<0.05)。(6)凋亡：与PBS组比较,CSE组与CSE+Adenovirus组DI增高,差异有统计学意义(p<0.05)；与CSE组比较,CSE+Adenovirus组DI显著增高,差异有统计学意义(p<0.05)。(7)增殖：与PBS组比较,CSE组与CSE+Adenovirus组肺组织Ki67表达增多,PI增高,差异有统计学意义(p0.05)。与CSE组比较,CSE+Adenovirus组PI无显著差别(p>0.05)。结论：CSE腹腔注射能建立肺气肿小鼠模型。腺病毒感染可促进CSE腹腔注射肺气肿小鼠肺部炎症反应,上调蛋白酶的表达,增加凋亡水平。更多还原

【Abstract】 Objective: To investigate the effect of adenovirus infection on the CSE(cigarette smoke extract) induced emphysema mice from the aspects of.pulmonary function, pathology, inflammation, protease, apoptosis and proliferation.Methods:BALB/c mice were injected with PBS/CSE at day1,11and22, and were received PBS/adenovirus intratracheally at day15. Mice were sacrificed at day28. Fluorescence of red fluorecent protein, pulmonary function, pathology of lung tissue by using mean linear intercept(MLI), destructive index(DI) and mean alveolar septal thickness(MAST), score of inflammation, expression of MMP-2and MMP-9, apoptosis and proliferation of lung tissue were measured.Results:(1) There was red fluorescence observed in CSE+Adenovirus group, while no red fluorescence in PBS and CSE group.(2) Compared with PBS group, airway resistance(RAW) was higher in CSE group and CSE+Adenovirus group(p<0.05), and lung dynamic compliance(Cdyn) was lower in CSE group and CSE+Adenovirus group(p<0.05). There were no significant differences in RAW and Cdyn between CSE group and CSE+Adenovirus group(p>0.05).(3) Compared with PBS group, MLI and DI were higher in CSE group and CSE+Adenovirus group(p<0.05), and MAST was lower in CSE group and CSE+Adenovirus group(p<0.05). There were no significant differences in MLI, DI and Cdyn between CSE group and CSE+Adenovirus group(p>0.05).(4) There were lots of inflammatory cells around small airway and microvessels in CSE+Adenovirus group. Compared with PBS group and CSE group, the scores of inflammation in small airway and microvessels were higher in CSE+Adenovirus group(p<0.05). There were no significant differences in the scores of inflammation in small airway and microvessels between PBS group and CSE group (p>0.05).(5) Compared with PBS group, the expression of MMP-2and MMP-9increased in CSE group and CSE+Adenovirus group(p<0.05). Compared with CSE group, the expression of MMP-2and MMP-9increased in CSE+Adenovirus group(p<0.05)(6) Compared with PBS group, the apoptosis index(DI) was higher in CSE group and CSE+Adenovirus group(p<0.05). Compared with CSE group, DI was higher in CSE+Adenovirus group(p<0.05).(7) Compared with PBS group, the proliferation index(PI) was higher in CSE group and CSE+Adenovirus group(p<0.05). There were no significant differences in PI between CSE group and CSE+Adenovirus group (p>0.05).Conclusions:CSE intraperitoneal injection can induce emphysema mice successfully. Adenovirus infection can promote the inflammation, increase protease expression and enhance apoptosis level.更多还原