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目的 研究γ干扰素(IFN-γ)对气道黏液细胞的作用及其机制。方法 (1)体外培养正常人气道上皮细胞 5~7d后,以含 50ng/ml的IFN γ的培养基再培养 3d,采用异硫氰酸荧光素(FITC)免疫荧光和末端脱氧核苷酸转移酶介导的脱氧三磷酸尿嘧啶缺口末端标记 (TUNEL)方法,观察IFN-γ诱导体外培养人气道黏液细胞凋亡及其机制。(2) 20只野生型C57BL/6J小鼠经致敏后按数字表法随机分为 4组,每组 5只。分别经鼻气道内滴注IFN-γ50ng、IFN-γ100ng、白细胞介素 13(IL-13)5μg,并以生理盐水作对照,采用细胞形态学方法,分别计数各组致敏小鼠单位气道长度内黏液细胞数;采用TUNEL法检测气道黏液细胞的凋亡改变。结果 ( 1 )免疫荧光细胞核染色联合TUNEL检测显示,经IFN γ作用后的人气道黏液细胞出现凋亡现象,细胞形态改变,胞核破裂;在细胞凋亡过程中,凋亡诱导因子Bax表达增加,并向线粒体转位。(2)经鼻向气管内滴注 100ngIFN γ后,小鼠气道单位长度(黏液细胞数 /毫米气道基底层,下同)黏液细胞数量为 (28±6)个 /mm,生理盐水组为(58±12)个 /mm,IL 13组为(59±6)个 /mm,生理盐水组和IL-13组与IFN γ100ng组比较差异有统计学意义(P<0.05);IFN γ50ng滴注组[ (48±11)个 /mm]与生理盐水组[ (58±12)个 /mm]和IL 13组[ (59±6)个
Objective To study the effect and mechanism of interferon-γ (IFN-γ) on airway mucous cells. Methods (1) Normal human airway epithelial cells were cultured in vitro for 5 to 7 days and then cultured for 3 days in culture medium containing 50 ng / ml IFN γ. FITC immunofluorescence and terminal deoxynucleotidyl transferase Enzyme-mediated deoxyuracil triphosphate nick end labeling (TUNEL) method to observe IFN-γ-induced apoptosis in human airway mucus cells and its mechanism. (2) Twenty wild-type C57BL / 6J mice were sensitized and divided into 4 groups randomly according to the digital table method, with 5 in each group. The nasal airway was instilled with IFN-γ50ng, IFN-γ100ng, IL-13 5μg, respectively, and normal saline as a control, using cell morphology method, were counted in each group of sensitized mouse airway Length of mucus cells; using TUNEL method to detect changes in airway mucus cells. Results (1) Immunofluorescence staining with nuclear staining and TUNEL assay showed that the apoptosis of mucous cells in human airways after IFNγ exposure was observed. The morphological changes and rupture of nuclei were observed. In the process of apoptosis, the expression of apoptosis-inducing factor Bax was increased , And translocate to mitochondria. (2) After intratracheal instillation of 100ng IFNγ, the number of mucous cells per unit length of airway (mucous cells / mm of airway basal layer, the same below) was (28 ± 6) mm / (59 ± 6) / mm for IL 13 group, and there was significant difference between the normal saline group and IL-13 group and the IFNγ100ng group (P <0.05) [(48 ± 11) / mm] compared with saline group [(58 ± 12) / mm] and IL13 group [(59 ± 6)