ACAP4-ezrin在HP VacA影响胃酸分泌中的作用及与慢性胃炎证候演变的相关性研究
发布时间：2018-07-16 18:08
【摘要】：目的:VacA是幽门螺杆菌重要的毒力因子之一,本团队已经进行的部分细胞实验表明其能够通过介导细胞外钙内流促进重要骨架结合蛋白ezrin的降解,进而影响ACAP4和质子泵在胃壁细胞膜上的定位,导致胃酸分泌能力的下调。为了验证这一结论的正确性以及进一步揭示ACAP4-ezrin信号通路在胃酸分泌功能中的作用,我们将研究思路由细胞学水平延伸至整体动物水平,利用ACAP4壁细胞特异性敲除小鼠研究了 HP VacA对胃酸分泌的影响并探讨其机制。同时我们对临床慢性胃炎患者进行了证候学调查,分析了慢性胃炎病理演变过程中证候的变化趋势,并利用从临床取得的患者胃黏膜组织探讨了 ACAP4-ezrin、质子泵表达与疾病病理演变以及证候分布特点和演变趋势之间的相关性。方法:本研究分为实验研究和临床研究两部分。第一部分为实验研究,本部分研究利用幽门螺杆菌VacA毒素对ACAP4基因剔除小鼠的胃酸分泌能力、胃粘膜对VacA易感性、VacA对ACAP4-ezrin及质子泵表达等的影响进行了研究,探讨了 ACAP4-ezrin在胃酸分泌当中的重要作用以及VacA对该通路的影响。第二部分为临床研究,本研究共收集了 300例慢性胃炎患者的四诊信息及其中59例患者的胃黏膜标本,运用描述性分析、因子分析等探讨慢性胃炎患者的证候特点及随着病理演变过程而出现的证候演变趋势,并结合免疫组化等研究方法,探索了 ACAP4-ezrin、质子泵等的表达与疾病的病理演变、证候演变的相关性。结果:①ACAP4KO小鼠生理状态下胃酸分泌能力降低,VacA能够抑制野生型小鼠的胃酸分泌功能,而对ACAP4 KO小鼠的酸分泌功能则基本无影响,同时ACAP4KO小鼠胃泌素介导的胃酸分泌水平出现代偿性升高;②野生型小鼠粘膜厚度变化以及腺体结构松散程度均较ACAP4 KO小鼠的损害较轻;给予VacA灌胃的ACAP4KO小鼠出现粘膜结构紊乱,局部出血等病理损害的几率明显高于野生型小鼠,并且上皮细胞出现空泡化的几率要明显高于野生型小鼠;③VacA还能够明显的减弱正常小鼠胃粘膜ACAP4、ezrin、质子泵的表达;④非萎缩性胃炎患者的证候类型以实证居多,萎缩性胃炎则以虚实夹杂证居多,癌前病变患者则以虚证居多,三者证候类型之间呈现了实证→虚实夹杂证→虚证的演变趋势;⑤ACAP4-ezrin及质子泵的表达与HP感染存在着明显的相关性,但ACAP4-ezrin及质子泵的表达与慢性胃炎患者的证候分布及其演变趋势之间暂未发现明显的相关性。结论:①ACAP4-ezrin在壁细胞胃酸分泌过程中承担着重要的作用,ACAP4-ezrin介导的胃酸分泌可以因为ACAP4的敲除而被阻断;②VacA破坏ezrin在壁细胞顶膜的定位及ACAP4-ezrin的相互作用在整体动物水平依然具有导致胃酸分泌减少的重要作用。但实验结果同时也提示,除了 ACAP4-ezrin介导的胃酸分泌途径外,依然存在着其他的介导胃酸分泌的途径或者通路,当ACAP4-ezrin的相互作用被阻断时,这些通路可能能够起到代偿的作用;③ACAP4基因敲除后,小鼠胃粘膜对VacA导致的粘膜损伤的易感性增加,提示ACAP4-ezrin可能是HP VacA作用于胃粘膜上皮细胞导致出现生理功能紊乱和病理损害的重要作用靶点之一;④实验研究和临床研究的结果表明,HP感染过程中分泌的VacA能够作用于ACAP4-ezrin复合体,通过破坏ezrin结构、ACAP4-ezrin相互作用以及抑制H,KATPase表达等多种方式抑制胃酸的分泌,这可能是HP感染的患者出现胃酸分泌过少的重要机制之一;⑤在慢性胃炎的病理演变过程中ACAP4、ezrin以及质子泵的表达随着胃粘膜结构破坏的加重逐渐减弱,部分统计学结果说明ACAP4、ezrin和质子泵的表达与慢性胃炎病情程度之间具有一定的相关性。因而ACAP4-ezrin以及质子泵不仅在胃粘膜壁细胞中正常的表达是决定胃酸分泌正常与否的重要指标,同时也有可能是提示胃粘膜病变严重程度的重要指标;⑥非萎缩性胃炎患者临床证候类型以实证居多,萎缩性胃炎则以虚实夹杂证居多,癌前病变患者则以虚证居多,三者证候之间呈现了实证→虚实夹杂证→虚证的演变趋势;⑦ACAP4-ezrin及质子泵的表达与HP感染存在着明显的相关性,但并不是绝对的负相关关系;而ACAP4-ezrin及质子泵的表达与慢性胃炎患者的证候分布及其演变趋势之间暂未发现明显的相关性。
[Abstract]:Objective: VacA is one of the important virulence factors of Helicobacter pylori. Some cell experiments already carried out by our team have shown that it can promote the degradation of important cytoskeleton binding protein ezrin by mediating extracellular calcium influx, thereby affecting the localization of ACAP4 and proton pumps on the membrane of the gastric wall, leading to the downregulation of gastric acid secretion. The validity of the conclusion and the further revelation of the role of ACAP4-ezrin signaling pathway in gastric acid secretion. We will study the effect of HP VacA on the secretion of gastric acid from the cytological level to the whole animal level, and explore the mechanism of the effect of HP VacA on gastric acid secretion. The patients with inflammation were investigated, and the trend of syndrome in the pathological process of chronic gastritis was analyzed. The correlation between ACAP4-ezrin, the expression of proton pump and pathological changes of the disease, the characteristics of syndrome distribution and the trend of evolution were discussed. The two part of the clinical study. The first part is the experimental study. This part studies the gastric acid secretion ability of ACAP4 gene knockout mice with Helicobacter pylori VacA toxin, the susceptibility to VacA in the gastric mucosa, the influence of VacA on the expression of ACAP4-ezrin and proton pump, and the important role of ACAP4-ezrin in the secretion of gastric acid and Va. The effect of cA on this pathway. The second part is clinical study. In this study, 300 cases of chronic gastritis were collected and 59 cases of gastric mucosa were collected. Descriptive analysis and factor analysis were used to investigate the syndrome characteristics of chronic gastritis patients and the trend of syndrome evolution with the process of pathological evolution. The results of the correlation between the expression of ACAP4-ezrin, the expression of proton pump and the pathological changes of the disease and the evolution of syndromes were explored. Results: (1) the secretion of gastric acid decreased in the physiological state of ACAP4KO mice, and the secretion function of gastric acid in wild type mice was inhibited by VacA, but the acid secretion function of ACAP4 KO mice was not affected, and AC The level of gastric acid secretion mediated by gastrin in AP4KO mice showed a compensatory increase, and the changes of mucosal thickness and loose structure of the wild type mice were lighter than that of ACAP4 KO mice; the risk of pathological damage such as mucosal structure disorder and local bleeding in the ACAP4KO mice given to VacA was significantly higher than that in the wild type mice, and the risk of pathological damage was significantly higher than that of the wild type mice. The probability of vacuolization of epithelial cells was significantly higher than that of wild type mice; (3) VacA could obviously weaken the expression of ACAP4, ezrin and proton pump in normal mice gastric mucosa; 4. The syndrome types of non atrophic gastritis were mostly positive, the atrophic gastritis was mostly false and solid, the precancerous lesions were mostly deficiency syndrome, and the three syndrome Between the types of syndrome and deficiency syndrome, there is an obvious correlation between the expression of ACAP4-ezrin and proton pump and HP infection, but the expression of ACAP4-ezrin and proton pump is not significantly correlated with the syndrome distribution and evolution trend of chronic gastritis. Conclusion: (1) ACAP4-ezrin Parietal cells play an important role in the secretion of gastric acid. ACAP4-ezrin mediated gastric acid secretion can be blocked by ACAP4 knockout; 2. VacA destruction of Ezrin in the apical membrane and the interaction of ACAP4-ezrin still plays an important role in the decrease of gastric acid secretion at the whole animal level. It is suggested that there are other pathways or pathways that mediate the secretion of gastric acid in addition to ACAP4-ezrin mediated gastric acid secretion. When the interaction of ACAP4-ezrin is blocked, these pathways may play a compensatory role. (3) after the ACAP4 knockout, the susceptibility to mucous membrane damage caused by VacA in the mouse gastric mucus is increased. ACAP4-ezrin may be one of the important targets for HP VacA to induce physiological dysfunction and pathological damage in gastric epithelial cells. (4) the results of experimental and clinical studies show that the VacA secreted in the HP infection can act on the ACAP4-ezrin complex, through the destruction of the ezrin structure, the interaction of ACAP4-ezrin and the interaction of the ACAP4-ezrin. Inhibition of H, KATPase expression and other ways to inhibit the secretion of gastric acid, which may be one of the important mechanisms of gastric acid secretion in patients with HP infection. 5. During the pathological process of chronic gastritis, the expression of ACAP4, ezrin and proton pump gradually weakened with the aggravation of gastric mucosal structure destruction, and some statistical results indicated ACAP4, ezrin There is a correlation between the expression of proton pump and the degree of chronic gastritis. Therefore, the normal expression of ACAP4-ezrin and proton pump not only in the gastric mucosa wall cells is an important index to determine the normal or not of gastric acid secretion, but also an important indicator of the severity of gastric mucosal lesions; 6. The types of clinical syndromes were mostly positive, the atrophic gastritis was mostly false and solid, the precancerous lesions were mostly false evidence, and the three syndromes showed the evolution trend of empirical, virtual and false evidence, and the expression of ACAP4-ezrin and proton pump had obvious correlation with HP infection, but it was not absolute negative correlation. There was no significant correlation between the expression of ACAP4-ezrin and proton pump and the distribution and evolution trend of syndromes in patients with chronic gastritis.
【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R573.3
本文编号：2127205
[Abstract]:Objective: VacA is one of the important virulence factors of Helicobacter pylori. Some cell experiments already carried out by our team have shown that it can promote the degradation of important cytoskeleton binding protein ezrin by mediating extracellular calcium influx, thereby affecting the localization of ACAP4 and proton pumps on the membrane of the gastric wall, leading to the downregulation of gastric acid secretion. The validity of the conclusion and the further revelation of the role of ACAP4-ezrin signaling pathway in gastric acid secretion. We will study the effect of HP VacA on the secretion of gastric acid from the cytological level to the whole animal level, and explore the mechanism of the effect of HP VacA on gastric acid secretion. The patients with inflammation were investigated, and the trend of syndrome in the pathological process of chronic gastritis was analyzed. The correlation between ACAP4-ezrin, the expression of proton pump and pathological changes of the disease, the characteristics of syndrome distribution and the trend of evolution were discussed. The two part of the clinical study. The first part is the experimental study. This part studies the gastric acid secretion ability of ACAP4 gene knockout mice with Helicobacter pylori VacA toxin, the susceptibility to VacA in the gastric mucosa, the influence of VacA on the expression of ACAP4-ezrin and proton pump, and the important role of ACAP4-ezrin in the secretion of gastric acid and Va. The effect of cA on this pathway. The second part is clinical study. In this study, 300 cases of chronic gastritis were collected and 59 cases of gastric mucosa were collected. Descriptive analysis and factor analysis were used to investigate the syndrome characteristics of chronic gastritis patients and the trend of syndrome evolution with the process of pathological evolution. The results of the correlation between the expression of ACAP4-ezrin, the expression of proton pump and the pathological changes of the disease and the evolution of syndromes were explored. Results: (1) the secretion of gastric acid decreased in the physiological state of ACAP4KO mice, and the secretion function of gastric acid in wild type mice was inhibited by VacA, but the acid secretion function of ACAP4 KO mice was not affected, and AC The level of gastric acid secretion mediated by gastrin in AP4KO mice showed a compensatory increase, and the changes of mucosal thickness and loose structure of the wild type mice were lighter than that of ACAP4 KO mice; the risk of pathological damage such as mucosal structure disorder and local bleeding in the ACAP4KO mice given to VacA was significantly higher than that in the wild type mice, and the risk of pathological damage was significantly higher than that of the wild type mice. The probability of vacuolization of epithelial cells was significantly higher than that of wild type mice; (3) VacA could obviously weaken the expression of ACAP4, ezrin and proton pump in normal mice gastric mucosa; 4. The syndrome types of non atrophic gastritis were mostly positive, the atrophic gastritis was mostly false and solid, the precancerous lesions were mostly deficiency syndrome, and the three syndrome Between the types of syndrome and deficiency syndrome, there is an obvious correlation between the expression of ACAP4-ezrin and proton pump and HP infection, but the expression of ACAP4-ezrin and proton pump is not significantly correlated with the syndrome distribution and evolution trend of chronic gastritis. Conclusion: (1) ACAP4-ezrin Parietal cells play an important role in the secretion of gastric acid. ACAP4-ezrin mediated gastric acid secretion can be blocked by ACAP4 knockout; 2. VacA destruction of Ezrin in the apical membrane and the interaction of ACAP4-ezrin still plays an important role in the decrease of gastric acid secretion at the whole animal level. It is suggested that there are other pathways or pathways that mediate the secretion of gastric acid in addition to ACAP4-ezrin mediated gastric acid secretion. When the interaction of ACAP4-ezrin is blocked, these pathways may play a compensatory role. (3) after the ACAP4 knockout, the susceptibility to mucous membrane damage caused by VacA in the mouse gastric mucus is increased. ACAP4-ezrin may be one of the important targets for HP VacA to induce physiological dysfunction and pathological damage in gastric epithelial cells. (4) the results of experimental and clinical studies show that the VacA secreted in the HP infection can act on the ACAP4-ezrin complex, through the destruction of the ezrin structure, the interaction of ACAP4-ezrin and the interaction of the ACAP4-ezrin. Inhibition of H, KATPase expression and other ways to inhibit the secretion of gastric acid, which may be one of the important mechanisms of gastric acid secretion in patients with HP infection. 5. During the pathological process of chronic gastritis, the expression of ACAP4, ezrin and proton pump gradually weakened with the aggravation of gastric mucosal structure destruction, and some statistical results indicated ACAP4, ezrin There is a correlation between the expression of proton pump and the degree of chronic gastritis. Therefore, the normal expression of ACAP4-ezrin and proton pump not only in the gastric mucosa wall cells is an important index to determine the normal or not of gastric acid secretion, but also an important indicator of the severity of gastric mucosal lesions; 6. The types of clinical syndromes were mostly positive, the atrophic gastritis was mostly false and solid, the precancerous lesions were mostly false evidence, and the three syndromes showed the evolution trend of empirical, virtual and false evidence, and the expression of ACAP4-ezrin and proton pump had obvious correlation with HP infection, but it was not absolute negative correlation. There was no significant correlation between the expression of ACAP4-ezrin and proton pump and the distribution and evolution trend of syndromes in patients with chronic gastritis.
【学位授予单位】：北京中医药大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R573.3
【参考文献】
相关期刊论文 前1条
1 陈润花;刘敏;陈亮;师宁;余求祥;丁霞;;幽门螺杆菌相关性慢性胃炎中医证候分布特点文献研究[J];中华中医药杂志;2013年06期
，本文编号：2127205
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