【摘要】：目的:以食物過(guò)敏小鼠為動(dòng)物模型,通過(guò)灌喂雙歧桿菌完整肽聚糖(Whole Peptidoglycan,WPG),觀察其對(duì)食物過(guò)敏的治療效果以及對(duì)調(diào)節(jié)性T細(xì)胞的作用;探討雙歧桿菌防治變態(tài)反應(yīng)性疾病的可能機(jī)制,為益生菌及其生物活性成分的進(jìn)一步開(kāi)發(fā)利用提供依據(jù)。 方法:對(duì)象:無(wú)雞蛋喂養(yǎng)4～6周齡SPF級(jí)Balb/c雌鼠24只,隨機(jī)分為3組,每組8只。取其中2組建立食物過(guò)敏模型,分別為:OVA致敏陽(yáng)性對(duì)照組(OVA組)和OVA激發(fā)后灌喂雙歧桿菌WPG治療組(T組);同時(shí)設(shè)正常生理鹽水陰性對(duì)照組(C組)。方法:(1)觀察各組癥狀表現(xiàn);(2)HE染色觀察各組小腸組織形態(tài)學(xué)變化;(3)ELISA法檢測(cè)血清中OVA特異性IgE水平;(4)流式細(xì)胞術(shù)分析脾臟單個(gè)核細(xì)胞懸液中CD4+CD25+調(diào)節(jié)性T細(xì)胞的數(shù)量變化;(5)ELISA法檢測(cè)血清中IL-10和TGF-β水平。 結(jié)果:(1)雙歧桿菌WPG對(duì)OVA介導(dǎo)的速發(fā)型變態(tài)反應(yīng)有緩解作用,T組僅12. 5%(1/8)的小鼠出現(xiàn)腹瀉且腹瀉程度較OVA組明顯減輕,而OVA組小鼠87.5%(7/8)出現(xiàn)腹瀉;T組小鼠平均體重增長(zhǎng)高于OVA組(P0.05),HE染色可見(jiàn)小腸絨毛中上皮細(xì)胞局灶性壞死、脫落、固有層炎癥細(xì)胞浸潤(rùn)現(xiàn)象明顯改善,OVA特異性IgE水平明顯降低,差異有顯著性(P0.05);(2)OVA組與C組相比,小鼠脾臟單個(gè)核細(xì)胞懸液中Foxp3陽(yáng)性細(xì)胞(Foxp3+CD4+CD25+調(diào)節(jié)性T細(xì)胞)占CD4+CD25+T細(xì)胞含量顯著降低(P0.01),血清IL-10和TGF-β水平皆降低,差異皆有統(tǒng)計(jì)學(xué)意義(P0.05);(3)T組小鼠脾臟單個(gè)核細(xì)胞懸液中CD4+CD25+T細(xì)胞占CD4+T細(xì)胞含量高于OVA組,差異有統(tǒng)計(jì)學(xué)差異(P0.05),較C組正常小鼠差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05);T組小鼠脾細(xì)胞中Foxp3陽(yáng)性細(xì)胞(Foxp3+CD4+CD25+調(diào)節(jié)性T細(xì)胞)占CD4+CD25+T細(xì)胞含量與OVA組相比顯著增高(P0.01)。T組小鼠血清IL-10水平高于OVA組(P0.05);TGF-β水平顯著高于OVA組(P0.01)。 結(jié)論:(1)雙歧桿菌WPG與雙歧桿菌全菌一樣,對(duì)OVA致敏小鼠具有相同的治療作用:可以緩解小鼠急性發(fā)作癥狀,降低食物過(guò)敏發(fā)病率,減輕腸道炎癥病理改變;(2)雙歧桿菌WPG在治療食物過(guò)敏中增加CD4+CD25+調(diào)節(jié)性T細(xì)胞數(shù)量,刺激細(xì)胞因子IL-10和TGF-β分泌,調(diào)節(jié)Th1/Th2平衡。可能是雙歧桿菌防治食物過(guò)敏的機(jī)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.
【學(xué)位授予單位】：重慶醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類(lèi)號(hào)】：R392.12

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