發(fā)布時(shí)間：2018-04-15 11:34

  本文選題：乳酸菌 + 派伊爾結(jié)�。� 參考：《江南大學(xué)》2009年博士論文

【摘要】： 派伊爾結(jié)(Peyer’s patch,PP)是腸道黏膜免疫系統(tǒng)的重要部位,是乳酸菌進(jìn)入腸黏膜免疫誘導(dǎo)部位的主要通道。本課題研究乳酸菌表面性質(zhì)和黏附黏液、PP性能的關(guān)系,不同黏附性能的乳酸菌和強(qiáng)黏附菌株(三種狀態(tài):活、紫外滅活和熱滅活)的免疫調(diào)節(jié)作用,強(qiáng)黏附菌株的抗感染作用和調(diào)節(jié)PP基因表達(dá)的作用。這些研究有助于闡明乳酸菌調(diào)節(jié)黏膜免疫機(jī)制,也將為乳酸菌免疫調(diào)節(jié)制劑的開(kāi)發(fā)提供理論依據(jù)。 試驗(yàn)先用大鼠黏液、小鼠PP為黏附基質(zhì),研究23株細(xì)菌的黏附性,結(jié)果顯示,Lactobacillus plantarum Fn008、Lactobacillus acidophilus Fn037、Lactobacillus rhamnosus GG(LGG)和Salmonella Typhimurium為強(qiáng)黏附菌株。利用細(xì)菌黏附正十六烷、水接觸角法測(cè)定細(xì)菌表面疏水性,結(jié)果表明,23株細(xì)菌的表面疏水性與黏附性沒(méi)有相關(guān)性。利用紅細(xì)胞凝集試驗(yàn)分析乳酸菌細(xì)胞表面凝集素的表達(dá)情況,結(jié)果表明23株的細(xì)菌凝集性與其黏附性沒(méi)有相關(guān)性。利用細(xì)菌表面疏水性、表面電荷和液體表面張力計(jì)算細(xì)菌表面自由能,結(jié)果表明22株乳酸菌的表面自由能與其黏附PP性呈負(fù)相關(guān)。 為了研究乳酸菌黏附PP能力與免疫調(diào)節(jié)的關(guān)系,分別給小鼠連續(xù)3天灌胃四株黏附PP能力不同的乳酸菌(LGG、Fn008、Fn022和Fn036)以及Salmonella,檢測(cè)脾淋巴細(xì)胞增殖能力及其基因IL-12表達(dá)能力、腹腔和派伊爾結(jié)巨噬細(xì)胞吞噬能力、細(xì)胞因子(IL-4,IL-10,IFN-γ,TGF-β,IL-17)、黏膜免疫指標(biāo)sIgA、免疫相關(guān)基因(CD80,Mfge8,CXCL13和CD79a)及細(xì)菌易位,結(jié)果表明:乳酸菌(LGG、Fn008、Fn022和Fn036)黏附派伊爾結(jié)能力與其增強(qiáng)PP巨噬細(xì)胞吞噬活性及產(chǎn)生sIgA能力呈正相關(guān);乳酸菌(LGG、Fn008、Fn022和Fn036)可誘導(dǎo)抗原呈遞細(xì)胞特異基因(CD80和Mfge8)和B細(xì)胞趨化因子CXCL13表達(dá)上調(diào),同時(shí)可抑制B細(xì)胞特異性基因CD79a表達(dá),這種作用依賴(lài)于其黏附PP的能力;乳酸菌和Salmonella都誘導(dǎo)了Th1免疫;乳酸菌Fn008、Fn022、LGG和Fn036沒(méi)有破壞腸道黏膜屏障,而Salmonella卻破壞腸道黏膜屏障引起細(xì)菌顯著易位。 為了研究乳酸菌Fn008經(jīng)過(guò)不同處理后黏附PP能力與其免疫調(diào)節(jié)作用的關(guān)系,分別給小鼠灌胃活菌Fn008 (V-Fn008)、紫外滅活菌Fn008 (UV-Fn008)、熱滅活菌Fn008 (HK-Fn008)、Fn008+Salmonella和Salmonella,連續(xù)3天,檢測(cè)脾淋巴細(xì)胞增殖能力及其基因IL-12表達(dá)能力、腹腔和派伊爾結(jié)巨噬細(xì)胞吞噬能力、細(xì)胞因子(IL-4,IL-10,IFN-γ,TGF-β,IL-17)、黏膜免疫指標(biāo)sIgA、免疫相關(guān)基因(CD80,Mfge8,CXCL13和CD79a)及細(xì)菌易位,結(jié)果表明:V-Fn008、UV-Fn008和HK-Fn008黏附PP的能力與其增強(qiáng)PP巨噬細(xì)胞吞噬能力及產(chǎn)生sIgA的能力呈正相關(guān)。V-Fn008、UV-Fn008和HK-Fn008可誘導(dǎo)抗原呈遞細(xì)胞特異基因(CD80和Mfge8)和B細(xì)胞趨化因子CXCL13表達(dá),同時(shí)可抑制B細(xì)胞特異性基因CD79a表達(dá),這種作用依賴(lài)于其黏附派伊爾結(jié)的能力�？诜㑇-Fn008、UV-Fn008和HK-Fn008都誘導(dǎo)了Th1細(xì)胞免疫,且UV-Fn008和HK-Fn008誘導(dǎo)的Th1免疫反應(yīng)弱于V-Fn008;活菌Fn008能抑制Salmonell引起的細(xì)菌易位; 利用On-tools分析口服乳酸菌Fn008小鼠空腸派伊爾結(jié)顯著改變的基因,研究相關(guān)的顯著性通路,結(jié)果表明,口服乳酸菌Fn008引起空腸派伊爾結(jié)內(nèi)的響應(yīng)基因主要與免疫、黏膜屏障和其它通路(肌動(dòng)蛋白骨架的調(diào)節(jié)和自然殺傷性細(xì)胞介導(dǎo)的細(xì)胞毒性等)通路有關(guān)。采用定量PCR結(jié)果表明,乳酸菌(LGG、Fn008、Fn022、Fn036、UV-Fn008和HK-Fn008)可誘導(dǎo)派伊爾結(jié)黏著斑通路中Itgb1、lamaα3和Thbs2三個(gè)基因表達(dá)上調(diào),這種作用依賴(lài)于其黏附派伊爾結(jié)的能力;乳桿菌Fn008通過(guò)使黏著斑中的基因FAK表達(dá)下調(diào),而上調(diào)Itgb1、lamaα3、Itgb4、Thbs2基因表達(dá),從而抑制沙門(mén)氏菌引起的細(xì)菌易位。 總之,本研究結(jié)果表明,特定乳酸菌主要通過(guò)表面自由能介導(dǎo)的作用力黏附PP,其黏附PP性與其增強(qiáng)派伊爾結(jié)巨噬細(xì)胞吞噬活性及誘導(dǎo)sIgA產(chǎn)生的能力呈正相關(guān);乳酸菌可誘導(dǎo)派伊爾結(jié)黏著斑通路中Itgb1、lamaα3和Thbs2三個(gè)基因表達(dá)上調(diào)、可誘導(dǎo)抗原呈遞細(xì)胞特異基因CD80和Mfge8表達(dá)上調(diào),同時(shí)可抑制B細(xì)胞特異性基因CD79a表達(dá),這種作用依賴(lài)于其黏附派伊爾結(jié)的能力;乳桿菌Fn008通過(guò)使黏著斑中的基因FAK表達(dá)下調(diào),而上調(diào)Itgb1、Itgb4、lamaα3、Thbs2基因表達(dá),從而抑制沙門(mén)氏菌引起的細(xì)菌易位。
[Abstract]:In this study , we study the relationship between the surface properties of lactic acid bacteria and the adhesion viscosity , the relationship between the properties of PP , the effects of different adhesive properties on anti - infection and the regulation of the expression of PP gene . These studies help to elucidate the mechanism of the regulation of the mucosal immune system by lactic acid bacteria and provide the theoretical basis for the development of the immune regulation preparation of lactic acid bacteria .



The results showed that the surface hydrophobicity of 23 strains was not related to the adhesion . The results showed that the surface hydrophobicity , surface charge and surface tension of the 23 strains were not related to the adhesion . The results showed that the surface free energy of 22 strains of lactic acid bacteria was negatively correlated with the adhesion of bacteria .



The effects of lactobacillus ( LGG , Fn008 , Fn022 and Fn36 ) on cell - specific genes ( CD80 , Mfge8 , CXCL13 and CD79a ) and bacterial translocation were studied . The results showed that the ability of lactobacillus ( LGG , Fn008 , Fn022 and Fn36 ) to induce cell - specific genes ( CD80 , Mfge8 , CXCL13 and CD79a ) and bacterial translocation showed that lactic acid bacteria ( LGG , Fn008 , Fn022 and Fn36 ) induced Th1 immunity . Lactic acid bacteria ( LGG , Fn008 , Fn022 , LGG and Fn36 ) did not destroy intestinal mucosal barrier , while Salmonella damaged intestinal mucosal barrier to cause significant bacterial translocation .



V - Fn008 , UV - Fn008 and HK - Fn008 were able to induce cell specific genes ( CD80 and Mfge8 ) and B cell chemokine CXCL13 . The results showed that the ability of V - Fn008 , UV - Fn008 and HK - Fn008 to induce cell specific genes ( CD80 , Mfge8 , CXCL13 and CD79a ) and bacterial translocation showed that V - Fn008 , UV - Fn008 and HK - Fn008 were able to induce Th1 cell immunity , and the immune response induced by UV - Fn008 , UV - Fn008 and HK - Fn008 was weaker than V - Fn008 ; and the viable bacteria Fn008 inhibited the translocation of bacteria caused by Salmonell .



The results showed that the response gene of Fn008 , Fn008 , Fn022 , Fn36 , UV - Fn008 and HK - Fn008 , could induce up - regulation of Itgb1 , 偽3 and Thbs2 gene expression . The results showed that lactic acid bacteria ( LGG , Fn008 , Fn022 , Fn36 , UV - Fn008 and HK - Fn008 ) could induce the expression of Itgb1 , 偽3 and Thbs2 genes in the sticky spot , and the expression of Itgb1 , 偽3 , Itgb4 and Thbs2 genes was raised .



In conclusion , the results showed that specific lactic acid bacteria were mainly mediated by surface free - energy - mediated adhesion ( PP ) , and its adherence PP was positively correlated with its ability to enhance the phagocytic activity and the ability of sIgA production . The expression of specific gene CD80 and Mfge8 was regulated by lactic acid bacteria .

【學(xué)位授予單位】：江南大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2009
【分類(lèi)號(hào)】：R392

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 劉瓊;王春鳳;;乳酸菌免疫調(diào)節(jié)功能研究進(jìn)展[J];吉林畜牧獸醫(yī);2011年10期

相關(guān)碩士學(xué)位論文 前1條

1 邵悅;益生菌對(duì)ND免疫雛雞局部黏膜體液免疫及IL-7 mRNA表達(dá)的影響[D];東北農(nóng)業(yè)大學(xué);2011年

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本文編號(hào)：1753982