论文标题:兔圆小囊在抗原刺激下脑肠肽类物质及炎症因子的动态变化观察
Studies on the Dynamic Changes of Gut-brain Peptides and Inflammation Factors in Scacculus Rotundus of Rabbit Stimulated by Antigen
论文作者 靳红
论文导师 佘锐萍,论文学位 博士,论文专业 基础兽医学
论文单位 中国农业大学,点击次数 501,论文页数 197页File Size39424k
2005-06-01论文网 http://www.lw23.com/lunwen_1095377/ 圆小囊;免疫组织化学;DNES;超微结构;粘膜免疫
sacculus rotundus; immuno-histochemistry; DNES; ultrastructure; mucosal immunity
本研究以神经-内分泌-免疫网络学说为依据,运用免疫组织化学的方法,结合组织培养技术、免疫电镜技术等,对离体兔圆小囊受到大肠杆菌等4种细菌、内毒素、外毒素等抗原刺激后,其组织内的4种脑肠肽和6种炎症相关因子表达量的动态变化进行了系统观察研究。同时,观察了在体感染兔瘟病毒后圆小囊内脑肠肽和炎症相关因子的变化。结果如下: 1.离体圆小囊受葡萄球菌、大肠杆菌、巴氏杆菌细菌抗原刺激后,胃泌素在圆小囊淋巴滤泡部位的表达,有显著升高的变化。表明淋巴组织滤泡部的DNES细胞大量释放了胃泌素。而沙门氏菌刺激圆小囊后,胃泌素在圆小囊淋巴滤泡部位的表达量逐渐降低。可能由于沙门氏菌的免疫抑制,使得免疫细胞受抑制,进而通过神经-内分泌-免疫网络中局部相关信使和信号转导通路影响了DNES细胞产生胃泌素。而VEGF表达量的变化主要是在淋巴组织滤泡部位急剧降低。 2、NSE在离体圆小囊绒毛部位的表达量的变化最显著,受到葡萄球菌、大肠杆菌、以及内毒素和外毒素刺激后,均呈现显著升高的变化。而受巴氏杆菌、沙门氏菌刺激后引起的变化不明显。表明NSE主要是受到破坏后细胞从胞内释放。 3、受到细菌以及内毒素刺激后,NF在各部位表达量的变化均不显著。而受到外毒素刺激后,在圆小囊淋巴组织圆顶部位出现了显著降低。推断NF可能主要受神经-内分泌-免疫网络的宏观调控调节。 4、受到细菌后离体圆小囊ecNOS的表达变化不显著。但受到内毒素和外毒素刺激后,分别在淋巴组织滤泡部位出现极显著降低。 5、受到葡萄球菌、巴氏杆菌、内毒素和外毒素刺激后的离体圆小囊,CD54表达量的变化差异不显著。大肠杆菌和沙门氏菌只在局部引起CD54表达量急剧降低。提示,大肠杆菌和沙门氏菌侵入肠道粘膜组织时,首先通过降低CD54的表达量,以利于细菌的侵入。 6、受到葡萄球菌、巴氏杆菌、大肠杆菌刺激后的离体圆小囊,E-cadherin的表达量没有显著变化。而沙门氏菌引起了离体圆小囊绒毛部位E-cadherin的表达量的显著降低,表明沙门氏菌入侵肠道枯膜也降低了E-cadherin的表达量。内毒素、外毒素刺激引起了淋巴组织圆顶部位E-cadherin的表达量的急剧降低,说明毒素由于缺乏细菌的表面多糖等结构,不能引起绒毛部位的变化,但是可以直接影响免疫细胞和DNES细胞丰富的圆顶部位对E-cadherin表达。 活体感染兔瘟病毒后,P物质的表达量显著升高;Actin和CD54的表达量呈降低趋势;而TGF-β和E-cadherin的表达量变化不显著;超微结构观察发现,感染兔瘟病毒后圆小囊结构不完整,细胞破损严重。胃泌素免疫电镜细胞化学观察结果显示阳性细胞数量明显升高。此结果表明P物质和胃泌素在病毒感染引起的炎症反应过程中,有一定的作用。 上述结果表明,离体兔圆小囊受到不同抗原刺激后,6种炎症相关因子和4种脑肠肽均出现了增多的趋势,而P物质、胃泌素、NSE三种脑肠肽的反应更为剧烈,而且在反应时间上早于炎症相关因子。提示这些脑肠肽在局部黏膜免疫反应中可能起着先导作用,进而推断脑肠肽在圆小囊局部黏膜免疫反应中可能有一定的调节作用。此结果进一步验证了圆小囊具有一定的局部自主调节作用。
Using immuno-histochemical, tissue cultural and immuno-electron microscopic methods, based on nerve-endocrine-immune net, the dynamic changes of 4 brain-gut peptides and 6 inflammation factors of excised saccalus rotunds of rabbit stimulated by germs, LPS, exotoxin were observed. The results showed:1. After stimulated by Staphylococcus aureus , Escherichia xoli O_78, Pasturella multocida,the content of gastrin in follicle increased significantly. This showed that the DNES cells in follicle released amount of gastrin. While stimulated by Salmonella, the results were opposite to the former. Maybe the immune inhibition of Salmonella made immune cells inhibited and through nerve-immune-endocrine system, the gastrin and other signal messages released by DNES cells were affected. The changes of VEGF decreased dramatically in follicle.2. Stimulated by Staphylococcus aureus , Escherichia .coli O_78, LPS and exotoxin, the contents of NSE in excised SR changed most in epithelium. While stimulated by Pasturella multocida and Escherichia .coli O_78, the content of NSE changed not so dramatically. This showed NSE most released from cells when damaged by some antigens.3. After stimulated by germs and LPS, the contents of NF in SR had no significant difference between the experiment and control group. But after stimulated by exotoxin, the content of NF in dome decreased significantly. Reckoned that NF might be regulated by nerve-immune-endocrine system..4.After stimulated by germs, the changes of contents of ecNOS were not so dramatically. While after stimulated by LPS and exotoxin, those in follicle decreased significantly.5.After stimulated by Staphylococcus aureus, Pasturella multocida ,LPS and exotoxin, the contents of CD54 had no significant difference between the experiment and control group. After stimulated by Escherichia .coli O_78 and Pasturella multocida, the contents of CD54 decreased greatly in part of SR. This suggested that when these two germs invaded intestines, first through, the content of CD54 decreased for invasion of germs.6.After stimulated by Staphylococcus aureus, Pasturella multocida and Escherichia .coli O_78, the contents of E-cadherin had no significant difference between the experiment and control group. While stimulated by Salmonella, the content of E-cad in epithelium decreased significantly. This showed that when Salmonella invaded intestine, the content of E-cad also decreased. After stimulated by LPS and extoxin, the contents of E-cad in dome decreased dramatically. This suggested that toxin could not change the content of E-cad but change those in dome of SR, where there were amount of immune cells and DNES cells.Infected with RHDV, the contents of SP in SR increased significantly, while the contents of actin and CD54 decreased. Ultrastructurally, the structures of SR were changed and the cells were damaged greatly. The result of immuno-electron cytochemistry showed the number of positive reaction cells of gastrin increased significantly. This suggested that SP and gastrin may interplaywith the inflammation caused by virus.All the results showed that when excised SR stimulated by various antigens, these 6 inflammation factors and 4 gut-brain peptides all increased, while SP, gastrin and NSE changed more dramatically. And those gut-brain peptides changed earlier than those inflammation factors. This suggested that these three gut-brain peptides might have a leading function in local mucosal immunity and it deduced that gut-brain peptides might have some regulative function. These results proved furtherly that SR had some self- regulative functions.
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