ACAP4-ezrin在HP VacA影響胃酸分泌中的作用及與慢性胃炎證候演變的相關(guān)性研究
發(fā)布時(shí)間：2018-07-16 18:08
【摘要】：目的:VacA是幽門螺桿菌重要的毒力因子之一,本團(tuán)隊(duì)已經(jīng)進(jìn)行的部分細(xì)胞實(shí)驗(yàn)表明其能夠通過介導(dǎo)細(xì)胞外鈣內(nèi)流促進(jìn)重要骨架結(jié)合蛋白ezrin的降解,進(jìn)而影響ACAP4和質(zhì)子泵在胃壁細(xì)胞膜上的定位,導(dǎo)致胃酸分泌能力的下調(diào)。為了驗(yàn)證這一結(jié)論的正確性以及進(jìn)一步揭示ACAP4-ezrin信號(hào)通路在胃酸分泌功能中的作用,我們將研究思路由細(xì)胞學(xué)水平延伸至整體動(dòng)物水平,利用ACAP4壁細(xì)胞特異性敲除小鼠研究了 HP VacA對(duì)胃酸分泌的影響并探討其機(jī)制。同時(shí)我們對(duì)臨床慢性胃炎患者進(jìn)行了證候?qū)W調(diào)查,分析了慢性胃炎病理演變過程中證候的變化趨勢(shì),并利用從臨床取得的患者胃黏膜組織探討了 ACAP4-ezrin、質(zhì)子泵表達(dá)與疾病病理演變以及證候分布特點(diǎn)和演變趨勢(shì)之間的相關(guān)性。方法:本研究分為實(shí)驗(yàn)研究和臨床研究?jī)刹糠�。第一部分為�?shí)驗(yàn)研究,本部分研究利用幽門螺桿菌VacA毒素對(duì)ACAP4基因剔除小鼠的胃酸分泌能力、胃粘膜對(duì)VacA易感性、VacA對(duì)ACAP4-ezrin及質(zhì)子泵表達(dá)等的影響進(jìn)行了研究,探討了 ACAP4-ezrin在胃酸分泌當(dāng)中的重要作用以及VacA對(duì)該通路的影響。第二部分為臨床研究,本研究共收集了 300例慢性胃炎患者的四診信息及其中59例患者的胃黏膜標(biāo)本,運(yùn)用描述性分析、因子分析等探討慢性胃炎患者的證候特點(diǎn)及隨著病理演變過程而出現(xiàn)的證候演變趨勢(shì),并結(jié)合免疫組化等研究方法,探索了 ACAP4-ezrin、質(zhì)子泵等的表達(dá)與疾病的病理演變、證候演變的相關(guān)性。結(jié)果:①ACAP4KO小鼠生理狀態(tài)下胃酸分泌能力降低,VacA能夠抑制野生型小鼠的胃酸分泌功能,而對(duì)ACAP4 KO小鼠的酸分泌功能則基本無(wú)影響,同時(shí)ACAP4KO小鼠胃泌素介導(dǎo)的胃酸分泌水平出現(xiàn)代償性升高;②野生型小鼠粘膜厚度變化以及腺體結(jié)構(gòu)松散程度均較ACAP4 KO小鼠的損害較輕;給予VacA灌胃的ACAP4KO小鼠出現(xiàn)粘膜結(jié)構(gòu)紊亂,局部出血等病理?yè)p害的幾率明顯高于野生型小鼠,并且上皮細(xì)胞出現(xiàn)空泡化的幾率要明顯高于野生型小鼠;③VacA還能夠明顯的減弱正常小鼠胃粘膜ACAP4、ezrin、質(zhì)子泵的表達(dá);④非萎縮性胃炎患者的證候類型以實(shí)證居多,萎縮性胃炎則以虛實(shí)夾雜證居多,癌前病變患者則以虛證居多,三者證候類型之間呈現(xiàn)了實(shí)證→虛實(shí)夾雜證→虛證的演變趨勢(shì);⑤ACAP4-ezrin及質(zhì)子泵的表達(dá)與HP感染存在著明顯的相關(guān)性,但ACAP4-ezrin及質(zhì)子泵的表達(dá)與慢性胃炎患者的證候分布及其演變趨勢(shì)之間暫未發(fā)現(xiàn)明顯的相關(guān)性。結(jié)論:①ACAP4-ezrin在壁細(xì)胞胃酸分泌過程中承擔(dān)著重要的作用,ACAP4-ezrin介導(dǎo)的胃酸分泌可以因?yàn)锳CAP4的敲除而被阻斷;②VacA破壞ezrin在壁細(xì)胞頂膜的定位及ACAP4-ezrin的相互作用在整體動(dòng)物水平依然具有導(dǎo)致胃酸分泌減少的重要作用。但實(shí)驗(yàn)結(jié)果同時(shí)也提示,除了 ACAP4-ezrin介導(dǎo)的胃酸分泌途徑外,依然存在著其他的介導(dǎo)胃酸分泌的途徑或者通路,當(dāng)ACAP4-ezrin的相互作用被阻斷時(shí),這些通路可能能夠起到代償?shù)淖饔?③ACAP4基因敲除后,小鼠胃粘膜對(duì)VacA導(dǎo)致的粘膜損傷的易感性增加,提示ACAP4-ezrin可能是HP VacA作用于胃粘膜上皮細(xì)胞導(dǎo)致出現(xiàn)生理功能紊亂和病理?yè)p害的重要作用靶點(diǎn)之一;④實(shí)驗(yàn)研究和臨床研究的結(jié)果表明,HP感染過程中分泌的VacA能夠作用于ACAP4-ezrin復(fù)合體,通過破壞ezrin結(jié)構(gòu)、ACAP4-ezrin相互作用以及抑制H,KATPase表達(dá)等多種方式抑制胃酸的分泌,這可能是HP感染的患者出現(xiàn)胃酸分泌過少的重要機(jī)制之一;⑤在慢性胃炎的病理演變過程中ACAP4、ezrin以及質(zhì)子泵的表達(dá)隨著胃粘膜結(jié)構(gòu)破壞的加重逐漸減弱,部分統(tǒng)計(jì)學(xué)結(jié)果說明ACAP4、ezrin和質(zhì)子泵的表達(dá)與慢性胃炎病情程度之間具有一定的相關(guān)性。因而ACAP4-ezrin以及質(zhì)子泵不僅在胃粘膜壁細(xì)胞中正常的表達(dá)是決定胃酸分泌正常與否的重要指標(biāo),同時(shí)也有可能是提示胃粘膜病變嚴(yán)重程度的重要指標(biāo);⑥非萎縮性胃炎患者臨床證候類型以實(shí)證居多,萎縮性胃炎則以虛實(shí)夾雜證居多,癌前病變患者則以虛證居多,三者證候之間呈現(xiàn)了實(shí)證→虛實(shí)夾雜證→虛證的演變趨勢(shì);⑦ACAP4-ezrin及質(zhì)子泵的表達(dá)與HP感染存在著明顯的相關(guān)性,但并不是絕對(duì)的負(fù)相關(guān)關(guān)系;而ACAP4-ezrin及質(zhì)子泵的表達(dá)與慢性胃炎患者的證候分布及其演變趨勢(shì)之間暫未發(fā)現(xiàn)明顯的相關(guān)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.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R573.3
本文編號(hào)：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.
【學(xué)位授予單位】：北京中醫(yī)藥大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R573.3
【參考文獻(xiàn)】
相關(guān)期刊論文 前1條
1 陳潤(rùn)花;劉敏;陳亮;師寧;余求祥;丁霞;;幽門螺桿菌相關(guān)性慢性胃炎中醫(yī)證候分布特點(diǎn)文獻(xiàn)研究[J];中華中醫(yī)藥雜志;2013年06期
，本文編號(hào)：2127205
本文鏈接：http://sikaile.net/yixuelunwen/xiaohjib/2127205.html
最近更新
教材專著
