摘要
【目的】观察嗜酸乳杆菌完整肽聚糖(Whole peptidoglycan,WPG)对致敏脾淋巴细胞Th1/Th2及Treg/Th17平衡的体外调节作用。【方法】通过腹腔注射β-乳球蛋白(β-Lg)建立BALB/c小鼠牛乳过敏模型。造模成功后,分离致敏小鼠的脾淋巴细胞并分别与不同剂量的WPG共同孵育,酶联免疫法检测细胞上清液中抗体(总IgE和特异性IgE),Th1/Th2及Treg/Th17相关细胞因子(IFN-γ,IL-4,TGF-β,IL-17)水平,流式细胞术检测脾淋巴细胞中CD3+、CD4+和CD8+的百分含量,荧光定量PCR法检测过敏小鼠脾细胞中Th1/Th2及Treg/Th17相关转录因子T-bet、GATA-3、Foxp3和RORγt mRNA的表达量。【结果】WPG体外刺激致敏脾细胞后可显著抑制IgE的产生,上调CD3+、CD4+细胞数及CD4+/CD8+比值,下调Th2型因子(IL-4,GATA-3mRNA)和Th17型因子(IL-17,RORγt mRNA)的表达,且与过敏组相比,中、高剂量WPG的作用效果显著(P<0.05);另外,WPG体外作用还上调了Th1型因子(IFN-γ,T-bet mRNA)及Treg型因子(TGF-β,Foxp3 mRNA)的表达,且具有剂量依赖性。【结论】嗜酸乳杆菌WPG体外刺激可有效纠正致敏脾淋巴细胞的Th1/Th2及Treg/Th17失衡。
[Objective] Aim of this study was to investigate the effect of whole peptidoglycan （WPG） from Lactobacillus acidophilus on regulating Th1/Th2 and Treg/Th17 balance in sensitized splenocytes. [Methods] BALB/c mice were intraperitoneally injected with β-Lactoglobulin （β-Lg）, the sensitized splenocytes were stimulated in vitro with different doses of WPG. Supernatants were collected and measured total IgE and β-Lg-specific IgE, Th1/Th2 and Treg/Th17 related cytokines （IFN-γ, IL-4, TGF-β, IL-17） production by ELISA assay; the percentage of CD3^＋, CD4^＋, CD8^＋ subgroups in spleen cells were measured by flow cytometry, and the levels of T-bet, GATA-3, Foxp3 and RORγt mRNA were measured by RT-PCR. [Results] Compared with the allergy group, treatment with medium and high dose of WPG significantly decreased IL-17, RORγt mRNA response. In addition, the numbers of CD3^＋ and CD4^＋ cells and the ratio of the secretion of total IgE and β-Lg-specific IgE, Th2-type （IL-4, GATA-3 mRNA） and Th17-type （CD4^＋/CD8^＋）, Th1-type （IFN-γ, T-bet mRNA） and Treg-type （TGF-β, Foxp3 mRNA） response were increased in the splenocytes stimulated with WPG in a dose manner. [Conclusion] Treatment with WPG from Lactobacillus acidophilus could regulate the imbalance of Th1/Th2 and Treg/Th17 in allergic spleen cells.
出处
《微生物学通报》
CAS
CSCD
北大核心
2014年第7期1334-1341,共8页
Microbiology China
基金
乳品科学教育部重点实验室开放基金项目(No.2012KLDSOF-07)
黑龙江省教育厅骨干教师基金项目(No.1254G008)
