摘要
目的研究大黄素对布鲁菌侵染巨噬细胞免疫功能的调节，为探寻新的药物治疗布鲁菌病提供理论和实验依据。方法取BALB/c小鼠分离骨髓细胞，加入MG-CSF诱导分化培养，用小鼠F4/80抗体、CD11b抗体标记细胞，流式检测骨髓细胞分化为巨噬细胞（MΦ）的比例；通过MTT法检测不同浓度大黄素对小鼠MΦ存活率的影响，同时以多西环素为对照，比较两种药物的半抑制浓度（IC50）；用布鲁菌侵染巨噬细胞（MΦ和细菌比例为100∶1）培养4 h，然后加入大黄素作用后，培养1、6、12、24、48 h，通过菌落计数法检测大黄素对MΦ内布鲁菌存活的影响，然后用ELISA法检测MΦ培养上清中细胞因子TNF-α、IL-6、IFN-γ的含量。结果培养第8天，骨髓细胞诱导分化为巨噬细胞的比例为91.28%；大黄素的IC50（608.4 μg/ml）明显高于多西环素的IC50（225.5 μg/ml）；菌落计数结果：6、12、24、48 h大黄素组logCFU值均显著低于空白组，大黄素组间比较，24 h大黄素组的logCFU值最低，且低于多西环素组；ELISA结果显示，与对照组比较，6、12、24 h大黄素组TNF-α、IL-6、IFN -γ含量明显增加，24 h大黄素组TNF-α、IL-6、IFN-γ含量最高；而12 h大黄素组IFN-γ含量[（74.233±4.416） pg/ml]与24 h大黄素组[（78.328±8.932） pg/ml]比较，差异无统计学意义。结论大黄素能够通过刺激TNF-α、IL-6、IFN-γ表达量增加，增强巨噬细胞杀伤布鲁菌的能力。
ObjectiveTo study the immunoregulatory effects of emodin on macrophages during Brucella infection and to provide theoretical and experimental basis for developing new drugs to treat brucellosis.MethodsBone marrow cells were isolated from BALB/c mice and cultured with MG-CSF to induce differentiation. Flow cytometry was used to detect the differentiation of bone marrow cells into macrophages （MΦ） by using FITC-labeled mouse anti-F4/80 antibody and PE-labeled anti-CD11b antibody. MTT method was used to detect the influences of various concentrations of emodin on the survival rate of MΦ. Doxycycline was used as the control to compare half maximal inhibitory concentrations （IC50） of the two drugs. MΦ were cultured with Brucella at a ratio of 100∶1 for 4 h. MΦ and Brucella were further cultured for 1, 6, 12, 24 and 48 h after adding emodin. Effects of emodin on the survival of MΦ were analyzed by colony counting method. ELISA was performed to detect the levels of TNF-α, IL-6 and IFN-γ in culture supernatants.ResultsOn day 8 of culturing, 91.28% of bone marrow cells differentiated into macrophages. The IC50 of emodin （608.4 μg/ml） was significantly higher than that of doxycycline （225.5 μg/ml）. The logCFU values of emodin stimulation groups （6, 12, 24 and 48 h） were significantly lower than those of blank control groups. Among all emodin stimulation groups, the 24 h group had the lowest logCFU value, which was also lower than that of the doxycycline treatment group. The levels of TNF-α, IL-6 and IFN-γ in 6, 12 and 24 h emodin stimulations group increased significantly as compared with those of the corresponding control groups. The levels of TNF-α, IL-6 and IFN-γ peaked at 24 h of culturing Brucella-infected MΦ with emodin. No significant difference in IFN-γ level was found between the 12 and 24 h emodin stimulation groups [（74.233±4.416） pg/ml vs （78.328±8.932） pg/ml].ConclusionEmodin may enhance the ability of macrophages to kill Brucella through promoting the expression
作者
朱珠
陈泽慧
陈安林
董泽令
彭丹
周小仙
杨智芳
Zhu Zhu, Chen Ze- hui, Chen Anlin, Dong Zeling, Peng Dan, Zhou Xiaoxian, Yang Zhifang(Department of Laboratory Medicine, Affiliated Hospital of Zunyi Medical College, Zunyi 563003, China ; Department of Laboratory Medicine Zunyi Medical College, Zunyi 563000, China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2018年第3期199-204,共6页
Chinese Journal of Microbiology and Immunology
