舌下含服免疫治疗对哮喘小鼠嗜酸性粒细胞凋亡及Fas、Bcl-2蛋白表达的影响

EFFECTS OF SUBLINGUAL IMMUNOTHERAPY ON EOSINOPHIL APOPTOSIS AND FAS AND BCl-2 PROTEIN EXPRESSION IN LUNG TISSUES OF ASTHMATIC MICE

目的:研究舌下含服疫苗对哮喘小鼠肺组织嗜酸性粒细胞凋亡及Fas、Bcl-2蛋白表达的影响。方法:21只BALb/c小鼠随机分为正常组、哮喘组和舌下含服粉尘螨疫苗治疗组。对正常组小鼠不做任何处理,对哮喘组小鼠用尘螨提取液诱发哮喘以制作哮喘模型,治疗组则以尘螨抗原致敏后进行免疫治疗。应用免疫组化Envision法检测肺组织Fas和Bcl-2蛋白的表达,再以体视学方法测算肺组织Fas和Bcl-2蛋白阳性细胞的体密度(Vv)和数密度(Nv)。用TUNEL法检测肺组织嗜酸性粒细胞凋亡情况。结果:①3组小鼠肺组织均可见嗜酸性粒细胞凋亡,治疗组嗜酸性粒细胞凋亡指数显著高于哮喘组(p<0.01)。②肺组织Fas、Bcl-2蛋白阳性细胞的体视学测量,治疗组Fas蛋白阳性细胞的体密度与数密度均比哮喘组高,差别均有统计学意义(p<0.05)。③治疗组Bcl-2蛋白阳性细胞的体密度与数密度均比哮喘组低,差别均有统计学意义(p<0.05)。结论:舌下含服免疫治疗可通过调节凋亡基因Fas、Bcl-2在嗜酸性粒细胞上的表达及肺组织内其它细胞上的表达,进而促进嗜酸性粒细胞及肺内其它炎性细胞在哮喘气道的凋亡来减少气道炎性细胞浸润,控制气道慢性炎症,达到防治哮喘的作用。

OBJECTIVE： To observe effect on eosinophil apoptosis and expression of Fas, Bcl -2 in lung tissue by sublingual baccal immunotherapy. METHOD： 21 BALb/c mice were divided into three groups randomly, control group, asthma model group and sublingual baccal vaccine - treatment group. Mice in the model group were injected with Dermatophagoides culinae extract to induce asthma and then treated with sublingual buccal immunotherapy. Immunohistochemical Envision method was adopted to examine the expression of Fas and Bcl - 2 and stereological method to calculate the two indices volume density, numerical density of the positive staining cells. TUNEL technique was used to determine the apoptosis of eosinophils in the lung tissue. RESULTS ： The eosinophil apoptosis could be seen in mice lung tissues of the three groups and its index in the asthma group mice lung tissue was lower than that of the treated group （ P 〈0. 01 ）. The volume density （Vv） and numerical density （Nv） could be noticed in the posi- tively staining cell in lung tissues. In the control group, Vv and Nv of Fas positive cells were respectively （ 3.66±0. 35 ）×10^-2 μm^0 and （7.78±0.48）×10^-6μm^-3; in the asthma group, Vv （2.43±0.22）×10^-2μm^0 and Nv （5.76±0.43）×10^-6μm^-3 ; and in the treated group, Vv （4.69±0.28）×10^-2μm^0 and Nv （10.55±0.54）×10^-6μm^-3. Vv and Nv of the treated group were higher than those of the asthma group, and the statistical significance was found in their differences （ P 〈 0. 05 ）. In the control group, Vv and Nv of the Bcl-2 positive cell were respectively （2.37±0.12）×10^-2μm^0 and （7.26±0.45）×10^-6μm^-3; in the asthma group, Vv （4.08±0.31）×10^-2μm^0 and Nv（11.86±0.53）×10^-6μm^-3; and in the treated group, Vv （2.14 ±0. 26）×10^-2 μm^0 and Nv （5.94±0. 39）×10^-6μm^-3. Vv and Nv of the treat group were lower than those of the asthma group, and the statistical significance was found in their differences （P〈0. 05）.