發(fā)布時(shí)間：2018-01-30 15:42

  本文關(guān)鍵詞： 幽門螺桿菌 Th17 白介素-23 白介素-17　出處：《南華大學(xué)》2011年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:通過建立幽門螺桿菌(H. pylori)感染的C57BL/6小鼠模型,分析感染后不同時(shí)期小鼠胃組織IL-17、IL-23 mRNA及蛋白表達(dá)情況,以及脾臟單細(xì)胞懸液中Th17細(xì)胞應(yīng)答情況,并分析感染后胃炎的嚴(yán)重程度與Th17-IL-17應(yīng)答效應(yīng)的相關(guān)性,同時(shí)設(shè)立治療組檢測(cè)上述指標(biāo)的變化情況。為探討IL-23/IL-17信號(hào)通路在H. pylori感染中的作用奠定基礎(chǔ),為H. pylori誘發(fā)的胃炎的免疫防御及治療提供新的理論和實(shí)驗(yàn)依據(jù)。 方法: (1)對(duì)6～8周齡的C57BL/6小鼠進(jìn)行灌胃感染H. pylori,每隔48小時(shí)一次,共灌胃5次。在H. pylori感染后的4、8、12周分別處死小鼠,鑒定H. pylori感染小鼠模型是否成功。同時(shí)設(shè)立治療組,采用三聯(lián)療法對(duì)灌胃感染4周后的小鼠進(jìn)行治療; (2)對(duì)對(duì)照組、感染組、治療組小鼠分別取部分胃組織作H㤘E染色,顯微鏡下觀察胃組織炎癥變化; (3)采用Trizol法對(duì)胃組織及脾細(xì)胞提取RNA,RT-PCR檢測(cè)細(xì)胞因子IL-17、IL-23 mRNA的表達(dá)情況; (4) ELISA檢測(cè)胃組織勻漿上清中細(xì)胞因子IL-17、IL-23蛋白含量; (5)采用PMA+Ionomycin或H. pylori WCP對(duì)脾淋巴細(xì)胞進(jìn)行刺激培養(yǎng),流式細(xì)胞術(shù)檢測(cè)脾臟單細(xì)胞懸液中Th17細(xì)胞應(yīng)答情況。 結(jié)果: (1)成功建立了H. pylori感染小鼠模型,感染組小鼠胃黏膜炎癥程度隨感染時(shí)間的延長而不斷加重; (2)與對(duì)照組相比,H. pylori感染組小鼠的胃組織中IL-17、IL-23的表達(dá)在mRNA及蛋白水平均顯著增高,H. pylori感染組小鼠的脾淋巴細(xì)胞中Th17細(xì)胞比例顯著高于對(duì)照組小鼠的該比值; (3) H. pylori感染組小鼠IL-17、IL-23 mRNA及蛋白含量以及脾淋巴細(xì)胞中Th17細(xì)胞比例隨感染時(shí)間延長而增加; (4)治療組小鼠IL-17、IL-23表達(dá)量及脾淋巴細(xì)胞中Th17細(xì)胞比率,與治療前即感染后4周的小鼠相比較均有所下降; (5)感染后不同時(shí)期小鼠胃黏膜的炎癥程度與IL-17、IL-23的表達(dá)量存在正相關(guān)。 結(jié)論: (1) H. pylori感染后IL-17、IL-23表達(dá)均上調(diào); (2) H. pylori感染后可以誘導(dǎo)Th17細(xì)胞應(yīng)答; (3) H. pylori感染后胃炎程度與胃組織IL-17、IL-23含量存在正相關(guān)。
[Abstract]:Objective: to establish a C57BL / 6 mouse model of Helicobacter pylori (H.pylori) infection and analyze the gastric tissue IL-17 of mice at different stages after infection. The expression of IL-23 mRNA and protein, and the response of Th17 cells in spleen single cell suspension. The relationship between the severity of post-infection gastritis and the response of Th17-IL-17 was analyzed. At the same time, a treatment group was set up to detect the changes of the above indexes, which laid a foundation for the study of the role of IL-23/IL-17 signaling pathway in H. pylori infection. To provide a new theoretical and experimental basis for the immune defense and treatment of H. pylori induced gastritis. Methods: (1) six weeks old C57BL / 6 mice were infused with H.pylorius every 48 hours, 5 times after H. pylori infection. After 12 weeks, the mice were killed to determine whether the model of H. pylori infection was successful or not. At the same time, the treatment group was set up, and the mice were treated with triple therapy after 4 weeks of gavage infection. (2) for the control group, the infection group and the treatment group, some gastric tissues were taken from the mice for H? The changes of inflammation in gastric tissue were observed under microscope. Trizol method was used to detect the expression of IL-17 IL-23 mRNA in gastric tissues and spleen cells by RT-PCR. (4) ELISA was used to detect the content of IL-17 and IL-23 in the supernatant of gastric tissue homogenate. 5) splenic lymphocytes were stimulated by PMA Ionomycin or H. pylori WCP. The response of Th17 cells in single cell suspension of spleen was detected by flow cytometry. Results: 1) the model of H. pylori infection in mice was successfully established, and the degree of gastric mucosal inflammation in infected mice was aggravated with the prolongation of infection time. (2) compared with the control group, the expression of IL-17 and IL-23 in the gastric tissues of mice infected with H. pylori was significantly increased in the level of mRNA and protein. The percentage of Th17 cells in spleen lymphocytes of pylori infected mice was significantly higher than that of control mice. The contents of IL-17 IL-23 mRNA and protein and the percentage of Th17 cells in spleen lymphocytes increased with the prolongation of infection time in mice infected with H. pylori. (4) the expression of IL-17 IL-23 and the ratio of Th17 cells in spleen lymphocytes in the treatment group were significantly lower than those in the control group (4 weeks after infection). (5) there was a positive correlation between the inflammatory degree of gastric mucosa and the expression of IL-17 and IL-23 in mice at different stages after infection. Conclusion: 1) the expression of IL-17 and IL-23 were up-regulated after infection with H. pylori. 2) Th17 cells could be induced by H. pylori infection. 3) the degree of gastritis after H. pylori infection was positively correlated with the content of IL-17 and IL-23 in gastric tissue.
【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R378

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 ;基礎(chǔ)疫苗學(xué)會(huì)議報(bào)告[J];中國生物制品學(xué)雜志;1989年01期

2 韋敏 ,張?zhí)煊?B_(16)-BL_6黑素瘤細(xì)胞因子基因轉(zhuǎn)移的免疫生物學(xué)意義[J];國外醫(yī)學(xué).耳鼻咽喉科學(xué)分冊(cè);1995年01期

3 陳詩書;“瘤苗”——細(xì)胞因子轉(zhuǎn)導(dǎo)人腫瘤細(xì)胞[J];生物工程進(jìn)展;1999年04期

4 馮進(jìn)波,許曉群,魏海明,劉文濤;臍血細(xì)胞因子基因表達(dá)的免疫學(xué)意義[J];臨床輸血與檢驗(yàn);2002年01期

5 宋仕玲,龔作炯;細(xì)胞因子及其網(wǎng)絡(luò)對(duì)肝纖維化的作用[J];人民軍醫(yī);2003年10期

6 余丹鳳,沈盛暉,張庚,胡馬洪,周穎;醒腦靜注射液對(duì)重癥顱腦損傷患者細(xì)胞因子及肺部感染的影響[J];中國中西醫(yī)結(jié)合急救雜志;2005年05期

7 楊小進(jìn),毛勤生,周新澤;二硫代氨基甲酸吡咯烷預(yù)防腹腔粘連及其機(jī)制探討[J];第二軍醫(yī)大學(xué)學(xué)報(bào);2005年11期

8 楊曉靜;閆元奎;路強(qiáng);;翼狀胬肉發(fā)病機(jī)制的研究進(jìn)展[J];內(nèi)蒙古醫(yī)學(xué)院學(xué)報(bào);2006年04期

9 張s，

本文編號(hào)：1476592