H.pylori对十二指肠粘膜上皮细胞碳酸氢盐转运蛋白的影响

发布时间：2018-07-11 09:46

本文选题：H.pylori + 十二指肠粘膜上皮细胞　；参考：《遵义医学院》2014年硕士论文

【摘要】：目的：通过动物实验和体外实验研究H. pylori感染前后十二指肠粘膜上皮细胞碳酸氢盐转运蛋白（CFTR、SLC26A6、SLC26A3）的表达，探讨H. pylori感染影响十二指肠粘膜上皮细胞碳酸氢盐分泌的机制，进一步明确H. pylori相关性十二指肠溃疡的发病机制。方法：动物实验：选用6周龄C57BL/6J雄鼠，随机分为对照组和模型组，模型组予H.pylori菌株SS1（VacA+CagA+）、NCTC11637（VacA+CagA-）、NCTC11639（VacA-CagA+）分次灌胃，于感染后3天、1周、2周、4周、8周取小鼠胃，用RT-PCR及吉姆萨染色法鉴定是否成功感染，再取近端十二指肠，用RT-PCR测定十二指肠粘膜碳酸氢盐转运蛋白（CFTR、SLC26A3、SLC26A6）mRNA表达量的变化。体外实验：原代培养人正常十二指肠粘膜上皮细胞，培养成功后加入上述H.pylori菌株，共培养24小时，用RT-PCR及Western-Blot的方法测定十二指肠粘膜碳酸氢盐转运蛋白mRNA表达量的变化。结果：动物实验：小鼠模型胃粘膜吉姆萨染可见大量逗点状、短棒状紫蓝色小体，即为H.pylori；RT-PCR扩增出H.pylori16srRNA基因，鉴定H.pylori成功感染。RT-PCR方法检测小鼠感染H.pylori后CFTR、SLC26A6、SLC26A3的mRNA表达水平较对照组明显降低（P＜0.05）。体外实验：RT-PCR方法检测H.pylori菌株与十二指肠粘膜上皮细胞共培养24h后CFTR、SLC26A3、SLC26A6的mRNA表达水平较对照组明显降低（P＜0.05）。Western-Blot方法检测H.pylori菌株与十二指肠粘膜上皮细胞共培养24h后CFTR蛋白质表达水平较对照组明显降低（P＜0.05）。其中，同时含有cagA和vacA的H.pylori国际标准株SS1对CFTR、SLC26A6和SLC26A3的影响大而持久。结论：H.pylori感染影响了十二指肠粘膜上皮细胞碳酸氢盐转运蛋白的表达，从而影响十二指肠粘膜上皮细胞碳酸氢盐分泌。H.pylori对十二指肠粘膜上皮细胞碳酸氢盐转运蛋白的影响与其致病因子有关。
[Abstract]:Objective: to investigate the expression of bicarbonate transporter (CFT) in duodenal mucosal epithelial cells before and after H. pylori infection, and to explore the mechanism of the effect of H. pylori infection on the secretion of bicarbonate in duodenal mucosal epithelial cells. To further clarify the pathogenesis of H. pylori associated duodenal ulcer. Methods: six week-old C57BL / 6J male rats were randomly divided into two groups: control group and model group. The model group was treated with H.pylori strain SS1 (Vaca CagA) NCTC11637 (VacA CagA-) NCTC11639 (VacA-CagA). RT-PCR and Gimsa staining were used to determine whether the infection was successful or not. Then the proximal duodenum was collected. The expression of hydrogen carbonate transporter (CFTRC26A3) mRNA in duodenal mucosa was determined by RT-PCR. In vitro experiment: the normal human duodenal epithelial cells were cultured in vitro. The above H. pylori strains were added to the duodenal mucosal epithelial cells. The expression of bicarbonate transporter mRNA in duodenal mucosa was determined by RT-PCR and Western-Blot. Results: in animal experiment, a large number of comma-like, short rod-shaped purple blue bodies were found in the gastric mucosa of mouse model. The H.pylori 16s rRNA gene was amplified by RT-PCR. The expression of SLC26A6 and SLC26A3 mRNA was significantly lower in H. pylori infected mice than that in control group (P < 0.05). Expression of Helicobacter pylori (H.pylori) and duodenal mucosal epithelial cells in co-cultured for 24 h after 24 hours of co-culture of H. pylori strain and duodenal mucosal epithelial cells was detected by in vitro experiment: the mRNA expression level of CFTRC26A3HSLC26A6 was significantly lower than that of the control group (P < 0.05) .Western-Blot method was used to detect the expression of H.pylori in co-cultured duodenal epithelial cells for 24 hours. The expression of CFTR protein was significantly lower than that of control group (P < 0.05). Among them, the international standard strain SS1 containing both cagA and vacA had a great and lasting effect on CFTRN SLC26A6 and SLC26A3. Conclusion the expression of bicarbonate transporter in duodenal mucosal epithelial cells was affected by H. pylori infection. The effect of H. pylori on the bicarbonate transporter of duodenal mucosal epithelial cells is related to its pathogenic factors.
【学位授予单位】：遵义医学院
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R573.1

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