香烟烟雾暴露对哮喘大鼠树突状细胞共刺激分子CD_(80)、CD_(86)表达的影响及意义

Effect and significance of cigarette smoke exposure on the expression of CD_(80),CD_(86) by marrow-derived dendritic cells from asthmatics rat

目的通过研究香烟烟雾对支气管哮喘(简称哮喘)大鼠骨髓来源的树突状细胞(dendriticcells,DCs)表面共刺激分子CD80、CD86表达的影响,探讨吸烟加重哮喘的免疫学机制。方法雄性Wistar大鼠40只随机分为对照组、哮喘组、烟雾暴露组、哮喘+烟雾暴露组。建立动物模型,培养各组大鼠骨髓来源DCs及脾脏来源淋巴细胞,用流式细胞仪检测DCs表面共刺激分子CD80和CD86的表达。同时进行混合淋巴细胞反应,酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测细胞上清中IFN-γ和IL-4含量,MTT法检测淋巴细胞增殖活性。结果①哮喘+烟雾暴露组大鼠DCs表面共刺激分子CD80和CD86的表达低于哮喘组,差异有统计学意义(P值均<0.01)。哮喘+烟雾暴露组大鼠DCs表面共刺激分子CD80表达低于对照组,哮喘组大鼠DCs表面共刺激分子CD86表达高于对照组,差异有统计学意义(P值均<0.01)。②哮喘组大鼠DCs刺激同种异体淋巴细胞增殖能力高于对照组,差异有统计学意义(P<0.01)。哮喘+烟雾暴露组大鼠DCs刺激同种异体淋巴细胞增殖能力低于哮喘组和对照组,差异有统计学意义(P值均<0.01)。③烟雾暴露+哮喘组和哮喘组DCs的IFN-γ表达低于对照组,IL-4表达高于对照组,差异有统计学意义(P值均<0.01),烟雾暴露+哮喘组DCs的IFN-γ表达低于哮喘组,IL-4表达高于哮喘组,差异有统计学意义(P值均<0.01)。④DCs表面共刺激分子CD80与IFN-γ表达、淋巴细胞增殖活性呈显著正相关(分别r=0.659、0.394,P<0.01和P<0.05),与IL-4表达呈显著负相关(r=-0.421,P<0.01),共刺激分子CD86的表达与淋巴细胞增殖活性呈显著正相关(r=0.712,P<0.01)。结论香烟烟雾暴露可以降低哮喘大鼠DCs表面共刺激分子CD80、CD86的表达,进而减弱DCs活化淋巴细胞的能力,这可能在吸烟影响哮喘气道炎症和免疫平衡中发挥了一定作用。

Objective To study the effect of cigarette smoke exposure on the expression of CD80,CD86 and relationship with the activity and immune balance of lymphocytes by marrow-derived dendritic cells(DCs) from asthmatics rat.Methods Forty Wistar rats were randomly divided into control group,smoke exposure group,asthma group,asthma combined smoke exposure group.Established animal models,then rat bone marrow-derived DCs and spleen lymphocytes were cultivated,and mixed lymphocyte responses(MLR) was done.Collected cell and cell supernatant.The expression of CD80 and CD86 was determined by flow cytometric analysis,and the proliferations of lymphocytes was examined with MTT colorimetric assay.The expression of IFN-γ and IL-4 was determined by using the enzyme linked immunosorbent assay(ELISA).Results ① Compared with asthma group,the expression levels of CD80 and CD86 decreased significantly in asthma combined smoke exposure group(P<0.01,respectively).Compared with control group,the expression levels of CD80 decreased in asthma combined smoke exposure group significantly(P<0.01).②Compared with control group,the stimulating activity of DCs on allogeneic lymphocytes increased significantly in both asthma group and asthma combined smoke exposure group(P<0.01,respectively).Compared with asthma group,the stimulating activity of DCs on allogeneic lymphocytes decreased in asthma combined smoke exposure group significantly(P<0.01).③ Compared with the control group,the expression of IFN-γ decreased and the expression of IL-4 increased remarkably in both the cigarette smoke exposure combined asthma group and the asthma group(P<0.01,respectively).Compared with the asthma group,the expression of IFN-γ decreased and the expression of IL-4 increased remarkably in the cigarette smoke exposure combined asthma group(P<0.01,respectively).④ The expression levels of CD80 was positively correlated with IFN-γ and the activity of lymphocytes,and negatively correlated with IL-4.The expression levels of CD86 was positively correlated with the activity of lymphocytes.Conclusions Cigarette smoke exposure may inhibit expression of CD80 and CD86 by marrow-derived DCs and reduce DCs stimulating activity on allogeneic lymphocytes in asthmatics rat,which may play an important role in aggravating airway inflammation and immune balance in asthma.