乳酸菌黏附派伊尔结及免疫调节作用研究

发布时间：2018-04-15 11:34

本文选题：乳酸菌 + 派伊尔结　；参考：《江南大学》2009年博士论文

【摘要】： 派伊尔结(Peyer’s patch,PP)是肠道黏膜免疫系统的重要部位,是乳酸菌进入肠黏膜免疫诱导部位的主要通道。本课题研究乳酸菌表面性质和黏附黏液、PP性能的关系,不同黏附性能的乳酸菌和强黏附菌株(三种状态:活、紫外灭活和热灭活)的免疫调节作用,强黏附菌株的抗感染作用和调节PP基因表达的作用。这些研究有助于阐明乳酸菌调节黏膜免疫机制,也将为乳酸菌免疫调节制剂的开发提供理论依据。试验先用大鼠黏液、小鼠PP为黏附基质,研究23株细菌的黏附性,结果显示,Lactobacillus plantarum Fn008、Lactobacillus acidophilus Fn037、Lactobacillus rhamnosus GG(LGG)和Salmonella Typhimurium为强黏附菌株。利用细菌黏附正十六烷、水接触角法测定细菌表面疏水性,结果表明,23株细菌的表面疏水性与黏附性没有相关性。利用红细胞凝集试验分析乳酸菌细胞表面凝集素的表达情况,结果表明23株的细菌凝集性与其黏附性没有相关性。利用细菌表面疏水性、表面电荷和液体表面张力计算细菌表面自由能,结果表明22株乳酸菌的表面自由能与其黏附PP性呈负相关。为了研究乳酸菌黏附PP能力与免疫调节的关系,分别给小鼠连续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,检测脾淋巴细胞增殖能力及其基因IL-12表达能力、腹腔和派伊尔结巨噬细胞吞噬能力、细胞因子(IL-4,IL-10,IFN-γ,TGF-β,IL-17)、黏膜免疫指标sIgA、免疫相关基因(CD80,Mfge8,CXCL13和CD79a)及细菌易位,结果表明:乳酸菌(LGG、Fn008、Fn022和Fn036)黏附派伊尔结能力与其增强PP巨噬细胞吞噬活性及产生sIgA能力呈正相关;乳酸菌(LGG、Fn008、Fn022和Fn036)可诱导抗原呈递细胞特异基因(CD80和Mfge8)和B细胞趋化因子CXCL13表达上调,同时可抑制B细胞特异性基因CD79a表达,这种作用依赖于其黏附PP的能力;乳酸菌和Salmonella都诱导了Th1免疫;乳酸菌Fn008、Fn022、LGG和Fn036没有破坏肠道黏膜屏障,而Salmonella却破坏肠道黏膜屏障引起细菌显著易位。为了研究乳酸菌Fn008经过不同处理后黏附PP能力与其免疫调节作用的关系,分别给小鼠灌胃活菌Fn008 (V-Fn008)、紫外灭活菌Fn008 (UV-Fn008)、热灭活菌Fn008 (HK-Fn008)、Fn008+Salmonella和Salmonella,连续3天,检测脾淋巴细胞增殖能力及其基因IL-12表达能力、腹腔和派伊尔结巨噬细胞吞噬能力、细胞因子(IL-4,IL-10,IFN-γ,TGF-β,IL-17)、黏膜免疫指标sIgA、免疫相关基因(CD80,Mfge8,CXCL13和CD79a)及细菌易位,结果表明:V-Fn008、UV-Fn008和HK-Fn008黏附PP的能力与其增强PP巨噬细胞吞噬能力及产生sIgA的能力呈正相关。V-Fn008、UV-Fn008和HK-Fn008可诱导抗原呈递细胞特异基因(CD80和Mfge8)和B细胞趋化因子CXCL13表达,同时可抑制B细胞特异性基因CD79a表达,这种作用依赖于其黏附派伊尔结的能力。口服V-Fn008、UV-Fn008和HK-Fn008都诱导了Th1细胞免疫,且UV-Fn008和HK-Fn008诱导的Th1免疫反应弱于V-Fn008;活菌Fn008能抑制Salmonell引起的细菌易位; 利用On-tools分析口服乳酸菌Fn008小鼠空肠派伊尔结显著改变的基因,研究相关的显著性通路,结果表明,口服乳酸菌Fn008引起空肠派伊尔结内的响应基因主要与免疫、黏膜屏障和其它通路(肌动蛋白骨架的调节和自然杀伤性细胞介导的细胞毒性等)通路有关。采用定量PCR结果表明,乳酸菌(LGG、Fn008、Fn022、Fn036、UV-Fn008和HK-Fn008)可诱导派伊尔结黏着斑通路中Itgb1、lamaα3和Thbs2三个基因表达上调,这种作用依赖于其黏附派伊尔结的能力;乳杆菌Fn008通过使黏着斑中的基因FAK表达下调,而上调Itgb1、lamaα3、Itgb4、Thbs2基因表达,从而抑制沙门氏菌引起的细菌易位。总之,本研究结果表明,特定乳酸菌主要通过表面自由能介导的作用力黏附PP,其黏附PP性与其增强派伊尔结巨噬细胞吞噬活性及诱导sIgA产生的能力呈正相关;乳酸菌可诱导派伊尔结黏着斑通路中Itgb1、lamaα3和Thbs2三个基因表达上调、可诱导抗原呈递细胞特异基因CD80和Mfge8表达上调,同时可抑制B细胞特异性基因CD79a表达,这种作用依赖于其黏附派伊尔结的能力;乳杆菌Fn008通过使黏着斑中的基因FAK表达下调,而上调Itgb1、Itgb4、lamaα3、Thbs2基因表达,从而抑制沙门氏菌引起的细菌易位。
[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 , 伪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 , 伪3 and Thbs2 genes in the sticky spot , and the expression of Itgb1 , 伪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 .

【学位授予单位】：江南大学
【学位级别】：博士
【学位授予年份】：2009
【分类号】：R392

【引证文献】