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目的通过腺病毒载体介导小鼠γ干扰素(mIFNγ)在小鼠肺脏转基因表达,探讨其对卵白蛋白(OVA)诱导过敏小鼠模型的治疗作用及其机制。方法48只BALB/c小鼠,按随机数字表法分为7组,即阴性对照组(A组)、过敏模型组Ⅰ、Ⅱ、Ⅲ组(B、D和F组)、转基因治疗组Ⅰ、Ⅱ、Ⅲ组(C、E和G组)。除A组外,B、C、D、E、F、G各组在第0天和第5天经腹腔给予OVA(每只15μg)致敏,第12～14天经气道吸入0.5%的OVA雾化液激发(20ml/次),建立过敏模型。第15天C、E、G组小鼠经鼻滴入50μl带有mIFNγ基因的复制缺陷型腺病毒(AdCMVmIFNγ)悬液[每只5×109/空斑形成单位(PFU)],A、B、D、F组小鼠经鼻滴入50μl生理盐水作为对照。第18天处死A、B、C组小鼠,第21天处死D和E组小鼠,第25天处死F和G组小鼠,获取支气管肺泡灌洗液(BALF)进行细胞分类计数,其上清液用酶联免疫吸附法测定mIFNγ的浓度,右肺用4%多聚甲醛固定及苏木精伊红(HE)染色,左肺组织提取总RNA,用半定量逆转录聚合酶链反应(RTPCR)了解肺组织中白细胞介素4(IL4)、IL-5、IL-6、IL-10、IL1-2、IL13和IL18的表达。结果(1)经鼻给予AdCMVmIFN-γ后,BALF中可测得mIFNγ的高效表达,C组第3天为(729.0±104.7)pg/ml,E组第6天为(984.5±119.1)pg/ml,G组第10天为(310.6±59.7)pg/ml。(2)B、C、D、
Objective To investigate the therapeutic effect and mechanism of mIFNγ on mouse lung by adenoviral vector mediated transfection of mouse ovalbumin (OVA) -induced allergy. Methods 48 BALB / c mice were randomly divided into 7 groups according to random number table: negative control group (group A), allergic model group Ⅰ, Ⅱ and Ⅲ (group B, D and F), transgenic group Ⅰ , Ⅱ, Ⅲ group (C, E and G group). Groups A, B, C, D, E, F and G were sensitized intraperitoneally with OVA (15 μg each) on day 0 and day 5, and 0.5% OVA aerosol excitation (20ml / time), the establishment of allergic models. On Day 15, mice in Groups C, E and G were intranasally instilled with 50 μl of a replication-deficient adenovirus (AdCMVmIFNγ) suspension containing mIFNγ gene [5 × 10 9 / plaque forming unit (PFU)], A, B D, F mice were instilled with 50μl saline as control. Groups A, B and C mice were sacrificed on day 18, D and E mice were sacrificed on day 21 and F and G mice were sacrificed on day 25 to obtain bronchoalveolar lavage fluid (BALF) The concentration of mIFNγin supernatant was determined by enzyme-linked immunosorbent assay. The right lung was fixed with 4% paraformaldehyde and HE staining. The total RNA was extracted from the left lung tissue. Semiquantitative reverse transcription-polymerase chain reaction (RTPCR) to understand the expression of interleukin 4 (IL4), IL-5, IL-6, IL-10, IL1-2, IL13 and IL18 in lung tissue. Results (1) The high expression of mIFNγ was detected in BALF after nasal administration of AdCMV mIFN-γ. The day 3 of C group was (729.0 ± 104.7) pg / ml and that of E group was (984.5 ± 119.1) pg / ml in group G and (310.6 ± 59.7) pg / ml on day 10 in group G. (2) B, C, D,