HP感染儿童外周血单个核细胞及胃窦黏膜共刺激分子B7-1、B7-2与CD28的表达

发布时间：2018-05-12 02:04

本文选题：幽门螺杆菌 + 消化性溃疡　；参考：《苏州大学》2012年硕士论文

【摘要】：目的：比较幽门螺杆菌(Hp)感染(有／无消化性溃疡)及对照组患儿外周血单个核细胞(PBMC)及胃窦黏膜细胞B7-1、B7-2和CD28的表达水平,探讨幽门螺杆菌感染在消化性溃疡中的作用机理。方法：选择有消化道症状的患儿共60例,均做胃镜检查并予胃窦部位活检,同时取血标本。共分三组,溃疡组(A组)：有消化道症状,胃镜检查有消化性溃疡,Hp(+),20例；无溃疡组(B组)：有消化道症状,胃镜检查无消化性溃疡,Hp(+),20例;对照组(C组)：有消化道症状,胃镜检查基本正常,Hp(-),20例。用半定量逆转录聚合酶链反应(RT-PCR)和免疫组化分别测定PBMC、胃窦黏膜细胞B7-1、B7-2和CD28mRNA及胃窦黏膜细胞B7-1、B7-2和CD28蛋白的表达水平,比较不同组之间上述指标有无差异。最后采用线性回归分析法对B7-1、B7-2和CD28mRNA及相应蛋白表达之间进行相关性分析。结果：1、免疫组化染色显示B7-1、B7-2主要表达于胃窦黏膜上皮细胞及固有层单个核细胞,CD28主要表达于固有层单个核细胞。HP感染患儿(A组及B组)胃窦黏膜固有层中单个核细胞数量较C组增加。从表达强度看,A组及B组患儿胃窦粘膜B7-1、CD28蛋白较C组增加(P0.05),但在A组及B组之间比较无差异(P0.05);而三组患儿之间胃窦粘膜B7-2蛋白染色均无差异(P0.05)。 2、RT-PCR法显示,A组及B组患儿胃窦黏膜细胞中B7-1mRNA和CD28mRNA的表达均高于C组(P0.05),但在A组及B组之间上述两分子的表达比较无差异(P0.05)；A组、B组及C组之间胃窦黏膜细胞B7-2mRNA表达均无差异。 3、RT-PCR法显示,A组、B组及C组之间PBMC中B7-1、B7-2和CD28mRNA表达均无差异(P0.05)。 4、相关性分析显示,胃窦黏膜局部三种协同刺激分子在转录水平(RT-PCR)与对应的蛋白表达水平(免疫组化)上均存在着一定的正相关关系。结论：1、免疫组化显示,Hp感染后患儿胃窦黏膜中存在着B7-1、B7-2及CD28三种蛋白的表达,说明机体免疫系统有针对Hp产生特异性的免疫应答。 2、儿童Hp感染后在转录及蛋白水平均存在胃窦黏膜局部B7-1及CD28上调,而B7-2无明显变化,提示局部细胞免疫可能参与了感染后黏膜的损伤过程。 3、儿童Hp感染后PBMC中并未发现B7-1mRNA、B7-2mRNA及CD28mRNA的明显变化,说明至少在儿童期,免疫反应主要在胃窦黏膜局部,而对全身的影响相对不大。
[Abstract]:Objective: to compare the expression levels of peripheral blood mononuclear cells (PBMCs) and gastric antral mucosal cells (B7-1TB7-2 and CD28) in patients with Helicobacter pylori (HP) infection (with or without peptic ulcer) and to explore the mechanism of Helicobacter pylori infection in peptic ulcer. Methods: 60 children with digestive tract symptoms were examined by gastroscopy, biopsy of gastric antrum and blood samples were taken. The ulcer group was divided into three groups: group A, with digestive tract symptoms and gastroscopy with peptic ulcer HP (n = 20); group B, without ulceration, with digestive tract symptoms and gastroscopy with no peptic ulcer; group C, control group, with digestive tract symptoms. The gastroscopy was normal in 20 cases. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression levels of B7-1B7-2 and CD28mRNA in gastric antral mucosal cells, and the expression levels of B7-1OB7-2 and CD28 in gastric antral mucosal cells. Finally, linear regression analysis was used to analyze the correlation between B7-1, B7-2, CD28mRNA and the corresponding protein expression. Results the immunohistochemical staining showed that B7-1mB7-2 was mainly expressed in gastric antral mucosal epithelial cells and lamina propria mononuclear cells. CD28 was mainly expressed in lamina propria mononuclear cells (group A and group B). The number of cells in group C was higher than that in group C. Compared with group C, the expression intensity of B7-1 + CD28 protein in gastric antral mucosa of group A and group B was higher than that of group C, but there was no difference between group A and group B, but there was no difference in the staining of B7-2 protein between group A and group B. there was no difference in B7-2 protein staining between group A and group B (P 0.05). 2RT-PCR showed that the expression of B7-1mRNA and CD28mRNA in gastric antral mucosa was higher in group A and B than that in group C, but there was no difference between group A and group B in the expression of B7-2mRNA in gastric antral mucosa. There was no difference between group A and group B in the expression of B7-2mRNA in gastric antral mucosa. 3RT-PCR showed that there was no difference in the expression of B7-1, B7-B7-2 and CD28mRNA in PBMC between group A, group B and group C, and there was no difference between group A and group C in the expression of B7-1B7-2 and CD28mRNA. 4. The correlation analysis showed that there was a positive correlation between the transcription level of RT-PCR and the corresponding protein expression level (immunohistochemistry) of three kinds of co-stimulatory molecules in gastric antrum mucosa. Conclusion the expression of B7-1, B7-B7-2 and CD28 in antral mucosa of children with HP infection was detected by immunohistochemistry, indicating that the immune system has a specific immune response to HP. 2. B7-1 and CD28 were up-regulated in gastric antral mucosa after HP infection, but B7-2 had no obvious change, suggesting that local cellular immunity might be involved in the process of mucosal injury after infection. 3. No significant changes of B7-1mRNA-B7-2 mRNA and CD28mRNA were found in PBMC after HP infection in children, indicating that the immune response was mainly localized in gastric antral mucosa at least in childhood, but had little effect on the whole body.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.1

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