本文選題：H.pylori + 十二指腸粘膜上皮細(xì)胞　； 參考：《遵義醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：通過動物實驗和體外實驗研究H. pylori感染前后十二指腸粘膜上皮細(xì)胞碳酸氫鹽轉(zhuǎn)運蛋白（CFTR、SLC26A6、SLC26A3）的表達(dá)，探討H. pylori感染影響十二指腸粘膜上皮細(xì)胞碳酸氫鹽分泌的機制，進(jìn)一步明確H. pylori相關(guān)性十二指腸潰瘍的發(fā)病機制。 方法：動物實驗：選用6周齡C57BL/6J雄鼠，隨機分為對照組和模型組，模型組予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及吉姆薩染色法鑒定是否成功感染，再取近端十二指腸，用RT-PCR測定十二指腸粘膜碳酸氫鹽轉(zhuǎn)運蛋白（CFTR、SLC26A3、SLC26A6）mRNA表達(dá)量的變化。體外實驗：原代培養(yǎng)人正常十二指腸粘膜上皮細(xì)胞，培養(yǎng)成功后加入上述H.pylori菌株，共培養(yǎng)24小時，用RT-PCR及Western-Blot的方法測定十二指腸粘膜碳酸氫鹽轉(zhuǎn)運蛋白mRNA表達(dá)量的變化。 結(jié)果：動物實驗：小鼠模型胃粘膜吉姆薩染可見大量逗點狀、短棒狀紫藍(lán)色小體，即為H.pylori；RT-PCR擴增出H.pylori16srRNA基因，鑒定H.pylori成功感染。RT-PCR方法檢測小鼠感染H.pylori后CFTR、SLC26A6、SLC26A3的mRNA表達(dá)水平較對照組明顯降低（P＜0.05）。體外實驗：RT-PCR方法檢測H.pylori菌株與十二指腸粘膜上皮細(xì)胞共培養(yǎng)24h后CFTR、SLC26A3、SLC26A6的mRNA表達(dá)水平較對照組明顯降低（P＜0.05）。Western-Blot方法檢測H.pylori菌株與十二指腸粘膜上皮細(xì)胞共培養(yǎng)24h后CFTR蛋白質(zhì)表達(dá)水平較對照組明顯降低（P＜0.05）。其中，同時含有cagA和vacA的H.pylori國際標(biāo)準(zhǔn)株SS1對CFTR、SLC26A6和SLC26A3的影響大而持久。 結(jié)論：H.pylori感染影響了十二指腸粘膜上皮細(xì)胞碳酸氫鹽轉(zhuǎn)運蛋白的表達(dá)，從而影響十二指腸粘膜上皮細(xì)胞碳酸氫鹽分泌。H.pylori對十二指腸粘膜上皮細(xì)胞碳酸氫鹽轉(zhuǎn)運蛋白的影響與其致病因子有關(guān)。
[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.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R573.1

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