双歧杆菌完整肽聚糖治疗食物过敏小鼠及对调节性T细胞影响的研究
发布时间：2018-08-24 13:55
【摘要】：目的:以食物过敏小鼠为动物模型,通过灌喂双歧杆菌完整肽聚糖(Whole Peptidoglycan,WPG),观察其对食物过敏的治疗效果以及对调节性T细胞的作用;探讨双歧杆菌防治变态反应性疾病的可能机制,为益生菌及其生物活性成分的进一步开发利用提供依据。 方法:对象:无鸡蛋喂养4～6周龄SPF级Balb/c雌鼠24只,随机分为3组,每组8只。取其中2组建立食物过敏模型,分别为:OVA致敏阳性对照组(OVA组)和OVA激发后灌喂双歧杆菌WPG治疗组(T组);同时设正常生理盐水阴性对照组(C组)。方法:(1)观察各组症状表现;(2)HE染色观察各组小肠组织形态学变化;(3)ELISA法检测血清中OVA特异性IgE水平;(4)流式细胞术分析脾脏单个核细胞悬液中CD4+CD25+调节性T细胞的数量变化;(5)ELISA法检测血清中IL-10和TGF-β水平。 结果:(1)双歧杆菌WPG对OVA介导的速发型变态反应有缓解作用,T组仅12. 5%(1/8)的小鼠出现腹泻且腹泻程度较OVA组明显减轻,而OVA组小鼠87.5%(7/8)出现腹泻;T组小鼠平均体重增长高于OVA组(P0.05),HE染色可见小肠绒毛中上皮细胞局灶性坏死、脱落、固有层炎症细胞浸润现象明显改善,OVA特异性IgE水平明显降低,差异有显著性(P0.05);(2)OVA组与C组相比,小鼠脾脏单个核细胞悬液中Foxp3阳性细胞(Foxp3+CD4+CD25+调节性T细胞)占CD4+CD25+T细胞含量显著降低(P0.01),血清IL-10和TGF-β水平皆降低,差异皆有统计学意义(P0.05);(3)T组小鼠脾脏单个核细胞悬液中CD4+CD25+T细胞占CD4+T细胞含量高于OVA组,差异有统计学差异(P0.05),较C组正常小鼠差异无统计学意义(P0.05);T组小鼠脾细胞中Foxp3阳性细胞(Foxp3+CD4+CD25+调节性T细胞)占CD4+CD25+T细胞含量与OVA组相比显著增高(P0.01)。T组小鼠血清IL-10水平高于OVA组(P0.05);TGF-β水平显著高于OVA组(P0.01)。 结论:(1)双歧杆菌WPG与双歧杆菌全菌一样,对OVA致敏小鼠具有相同的治疗作用:可以缓解小鼠急性发作症状,降低食物过敏发病率,减轻肠道炎症病理改变;(2)双歧杆菌WPG在治疗食物过敏中增加CD4+CD25+调节性T细胞数量,刺激细胞因子IL-10和TGF-β分泌,调节Th1/Th2平衡。可能是双歧杆菌防治食物过敏的机制之一。
[Abstract]:Objective: to observe the therapeutic effect of bifidobacterium integrity peptidoglycan (Whole Peptidoglycan,WPG) on food allergy and its effect on regulatory T cells in mice with food allergy. To explore the possible mechanism of Bifidobacterium in the prevention and treatment of allergic diseases, and to provide a basis for the further development and utilization of probiotics and their bioactive components. Methods: Twenty-four SPF Balb/c female rats aged 4 weeks and 6 weeks without egg were randomly divided into 3 groups with 8 rats in each group. Food allergy models were established in two groups: OVA group (OVA group), WPG group (T group) fed with OVA, and normal saline negative control group (C group). Methods: (1) observe the symptoms of each group; (2) observe the changes of intestinal histomorphology by HE staining; (3) detect the level of OVA specific IgE in serum by ELISA method; (4) analyze the quantity of CD4 CD25 regulatory T cells in splenic mononuclear cell suspension by flow cytometry. (5) the levels of IL-10 and TGF- 尾 in serum were detected by ELISA method. Results: (1) Bifidobacterium WPG could alleviate the OVA mediated hypersensitivity in T group only 12.2%. 5% (1 / 8) of the mice developed diarrhea and the degree of diarrhea was significantly less than that of the OVA group, while 87.5% (7 / 8) of the mice in the OVA group showed a higher average weight gain than that in the OVA group (P0.05). The infiltration of inflammatory cells in lamina propria significantly improved the level of OVA-specific IgE, and the difference was significant (P0.05); (2) between OVA group and C group. Foxp3 positive cells (Foxp3 CD4 CD25 regulatory T cells) in splenic mononuclear cell suspension significantly decreased the content of CD4 CD25 T cells (P0.01), and the levels of serum IL-10 and TGF- 尾 were both decreased. The content of CD4 CD25 T cells in splenic mononuclear cell suspension of T group was significantly higher than that of OVA group (P0.05); (3). The difference was statistically significant (P0.05), but there was no significant difference between group C and group C (P0.05). The percentage of Foxp3 positive cells (Foxp3 CD4 CD25 regulatory T cells) in CD4 CD25 T cells in T group was significantly higher than that in OVA group (P0.01) .T group was significantly higher than that in OVA group (P0.01). The level of IL-10 was higher than that of OVA group (P0.05) and the level of TGF- 尾 was significantly higher than that of OVA group (P0.01). Conclusion: (1) Bifidobacterium WPG has the same therapeutic effect on OVA sensitized mice as the whole bifidobacterium strain: it can relieve the acute attack symptoms and reduce the incidence of food allergy in mice. (2) Bifidobacterium WPG increased the number of CD4 CD25 regulatory T cells, stimulated the secretion of cytokines IL-10 and TGF- 尾, and regulated Th1/Th2 balance in the treatment of food allergy. It may be one of the mechanisms of Bifidobacterium to prevent food allergy.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.12
本文编号：2201028
[Abstract]:Objective: to observe the therapeutic effect of bifidobacterium integrity peptidoglycan (Whole Peptidoglycan,WPG) on food allergy and its effect on regulatory T cells in mice with food allergy. To explore the possible mechanism of Bifidobacterium in the prevention and treatment of allergic diseases, and to provide a basis for the further development and utilization of probiotics and their bioactive components. Methods: Twenty-four SPF Balb/c female rats aged 4 weeks and 6 weeks without egg were randomly divided into 3 groups with 8 rats in each group. Food allergy models were established in two groups: OVA group (OVA group), WPG group (T group) fed with OVA, and normal saline negative control group (C group). Methods: (1) observe the symptoms of each group; (2) observe the changes of intestinal histomorphology by HE staining; (3) detect the level of OVA specific IgE in serum by ELISA method; (4) analyze the quantity of CD4 CD25 regulatory T cells in splenic mononuclear cell suspension by flow cytometry. (5) the levels of IL-10 and TGF- 尾 in serum were detected by ELISA method. Results: (1) Bifidobacterium WPG could alleviate the OVA mediated hypersensitivity in T group only 12.2%. 5% (1 / 8) of the mice developed diarrhea and the degree of diarrhea was significantly less than that of the OVA group, while 87.5% (7 / 8) of the mice in the OVA group showed a higher average weight gain than that in the OVA group (P0.05). The infiltration of inflammatory cells in lamina propria significantly improved the level of OVA-specific IgE, and the difference was significant (P0.05); (2) between OVA group and C group. Foxp3 positive cells (Foxp3 CD4 CD25 regulatory T cells) in splenic mononuclear cell suspension significantly decreased the content of CD4 CD25 T cells (P0.01), and the levels of serum IL-10 and TGF- 尾 were both decreased. The content of CD4 CD25 T cells in splenic mononuclear cell suspension of T group was significantly higher than that of OVA group (P0.05); (3). The difference was statistically significant (P0.05), but there was no significant difference between group C and group C (P0.05). The percentage of Foxp3 positive cells (Foxp3 CD4 CD25 regulatory T cells) in CD4 CD25 T cells in T group was significantly higher than that in OVA group (P0.01) .T group was significantly higher than that in OVA group (P0.01). The level of IL-10 was higher than that of OVA group (P0.05) and the level of TGF- 尾 was significantly higher than that of OVA group (P0.01). Conclusion: (1) Bifidobacterium WPG has the same therapeutic effect on OVA sensitized mice as the whole bifidobacterium strain: it can relieve the acute attack symptoms and reduce the incidence of food allergy in mice. (2) Bifidobacterium WPG increased the number of CD4 CD25 regulatory T cells, stimulated the secretion of cytokines IL-10 and TGF- 尾, and regulated Th1/Th2 balance in the treatment of food allergy. It may be one of the mechanisms of Bifidobacterium to prevent food allergy.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.12
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