【摘要】：門脈高壓性胃病(portal hypertensive gastropathy, PHG)是肝硬化門靜脈高壓的常見并發(fā)癥之一。其主要的臨床表現為上消化道出血，是肝硬化門脈高壓癥并發(fā)上消化道出血的主要原因之一，僅次于食管胃底靜脈曲張破裂引起的上消化道出血，具有重要的臨床意義。PHG患者會出現胃部微循環(huán)的異常，胃黏膜血流量減少，引起胃黏膜屏障損傷，使胃黏膜對各種損傷更加敏感，且抗損傷能力減弱。胃黏膜上皮細胞間的緊密連接是胃黏膜屏障的重要組成部分，其位于相鄰細胞間隙的頂端側面，具有維持細胞極性和調節(jié)滲透性兩個主要功能。對于維持胃黏膜屏障功能的穩(wěn)定具有非常重要的作用。與胃黏膜上皮細胞緊密連接(tight junction, TJ)相關的蛋白，主要包括閉鎖(Occludin)蛋白、Claudin蛋白、zonula occludens (ZOs)蛋白以及連接黏附分子(junction adhesion molecular, JAM)等。胃黏膜上皮細胞緊密連接蛋白表達量的降低，提示胃黏膜屏障功能受損。 鹽酸普萘洛爾是一種非選擇性β受體阻滯劑，是目前在治療門脈高壓中應用最為廣泛的藥物。它可以通過降低門靜脈血壓、減少胃黏膜血流量，從而起到治療PHG的作用。替普瑞酮是一萜類化合物，現廣泛應用于抗?jié)冎委�。目前研究認為，它可通過誘導熱休克蛋白(HSP)的表達和促進胃黏膜內源性前列腺素E(prostaglandin E, PGE)的合成與釋放來保護胃黏膜。 因此，本項研究的目的在于探討緊密連接相關蛋白在PHG患者胃黏膜屏障損傷中的作用，同時探討鹽酸普萘洛爾以及聯(lián)合替普瑞酮對PHG胃黏膜緊密連接蛋白表達的影響。 目的：本研究旨在揭示PHG胃黏膜屏障損傷的分子機制，并探討鹽酸普萘洛爾和替普瑞酮對PHG的治療效果及對胃黏膜上皮細胞緊密連接蛋白表達的影響。 方法：收集行胃鏡檢查的PHG患者10例。將其隨機分為兩組，在行胃鏡檢查及標本采集后，其中一組給予鹽酸普萘洛爾（30mg/天）治療，另一組給予鹽酸普萘洛爾（30mg/天）聯(lián)合替普瑞酮膠囊（150mg/天）治療，兩組患者均在治療1個月后復查胃鏡并再次采集標本，同時觀察患者內鏡下胃黏膜病變的變化情況。另外收集10例行上消化道內鏡檢查的健康體檢者作為健康對照組。于胃底體交界處口側3~5cm大彎側取材，應用免疫組織化學方法檢測各組胃黏膜上皮細胞間緊密連接蛋白Occludin和ZO-1的表達情況。 結果：PHG患者胃黏膜上皮細胞緊密連接蛋白Occludin和ZO-1表達減少。PHG患者治療前與健康對照組相比：①胃鏡下表現：PHG患者內鏡下可見到典型的馬賽克樣改變、蛇皮樣改變、散在的櫻桃紅樣斑點或彌漫性出血性改變等。②組織形態(tài)學表現：PHG患者黏膜層及黏膜下層可見明顯的毛細血管擴張，無明顯的炎癥改變。③Occludin蛋白和ZO-1蛋白的免疫組織化學結果：健康對照組Occludin蛋白及ZO-1蛋白均呈棕褐色線狀連續(xù)均勻分布于胃黏膜上皮細胞的細胞膜，PHG患者藥物干預前與健康對照組比較，Occludin蛋白及ZO-1蛋白染色均明顯變淺，不連續(xù)，有部分缺失，且分布也不均勻，Occludin的平均光密度值(optical density,OD)分別為：(0.03±0.01),(0.14±0.02),(P0.001), ZO-1的OD值分別為：(0.03±0.01),(0.16±0.02),(P0.001)，均有統(tǒng)計學差異。 鹽酸普萘洛爾可以增加PHG患者胃黏膜上皮細胞緊密連接蛋白Occludin和ZO-1的表達量。單用鹽酸普萘洛爾干預組，PHG患者治療后與治療前比較：①胃鏡下表現：治療后PHG患者胃黏膜病變較前好轉。②組織形態(tài)學表現：治療后PHG患者黏膜層及黏膜下層毛細血管擴張減輕。③Occludin蛋白和ZO-1蛋白的免疫組織化學結果：治療后的患者胃黏膜上皮細胞的Occludin蛋白和ZO-1蛋白染色均較前加深，仍可見部分缺失，但缺失較前減少，其中Occludin蛋白的OD值分別為：(0.06±0.01),(0.03±0.01),(P0.01), ZO-1蛋白的OD值分別為：(0.07±0.01),(0.03±0.01),(P0.01)，均有統(tǒng)計學差異。 聯(lián)合應用鹽酸普萘洛爾和替普瑞酮比單用鹽酸普萘洛爾對PHG患者胃黏膜上皮細胞緊密連接蛋白Occludin和ZO-1表達量的增加更明顯。聯(lián)合應用鹽酸普萘洛爾和替普瑞酮干預組治療后與單獨應用鹽酸普萘洛爾干預組治療后比較：①胃鏡下表現：聯(lián)合應用鹽酸普萘洛爾和替普瑞酮組比單用鹽酸普萘洛爾組胃黏膜病變好轉更加明顯。②組織形態(tài)學表現：聯(lián)合應用鹽酸普萘洛爾和替普瑞酮組比單用鹽酸普萘洛爾組黏膜層及黏膜下層毛細血管擴張減輕更加明顯。③Occludin蛋白和ZO-1蛋白的免疫組織化學結果：聯(lián)合應用鹽酸普萘洛爾和替普瑞酮治療后，PHG患者胃黏膜上皮細胞的Occludin蛋白和ZO-1蛋白染色加深更明顯，缺失部分減少更明顯，Occludin蛋白的OD值分別為：(0.11±0.01),(0.06±0.01),(P0.01)，，ZO-1蛋白的OD值分別為：(0.12±0.02),(0.07±0.01),(P0.01)，均有統(tǒng)計學差異。 結論：PHG患者胃黏膜上皮細胞緊密連接蛋白Occludin和ZO-1表達減少，這可能是PHG胃黏膜屏障損傷的重要原因，替普瑞酮可能通過上調Occludin和ZO-1的表達來保護胃黏膜屏障。
[Abstract]:Portal hypertensive gastropathy (PHG) is one of the common complications of portal hypertension in cirrhosis. The main clinical manifestation of PHG is upper gastrointestinal bleeding, which is one of the main causes of upper gastrointestinal bleeding in cirrhotic portal hypertension, second only to upper gastrointestinal bleeding caused by rupture of esophageal and gastric varices. PHG patients may have abnormal gastric microcirculation, decreased gastric mucosal blood flow, resulting in damage to the gastric mucosal barrier, making the gastric mucosa more sensitive to various kinds of damage, and weakening the ability to resist damage. On the apical side, it has two main functions: maintaining cell polarity and regulating permeability. It plays an important role in maintaining the stability of gastric mucosal barrier function. Proteins related to tight junction (TJ) of gastric mucosal epithelial cells, mainly including Occludin, Claudin, zonula occludens (ZOs), and so on. The expression of tight junction molecule (JAM) in gastric mucosal epithelial cells decreased, suggesting impaired gastric mucosal barrier function.
Propranolol hydrochloride is a non-selective beta-blocker, which is the most widely used drug in the treatment of portal hypertension. It can reduce portal blood pressure, gastric mucosal blood flow, and thus play a role in the treatment of PHG. Teprenone is a terpenoid compound and is widely used in anti-ulcer therapy. It can protect gastric mucosa by inducing the expression of heat shock protein (HSP) and promoting the synthesis and release of endogenous prostaglandin E (PGE).
Therefore, the purpose of this study was to investigate the role of tight junction related proteins in gastric mucosal barrier injury in PHG patients, and to investigate the effects of propranolol hydrochloride and teprenone on the expression of tight junction proteins in gastric mucosa of PHG patients.
AIM: To explore the molecular mechanism of gastric mucosal barrier injury in PHG and the effects of propranolol hydrochloride and teprenone on the expression of tight junction protein in gastric epithelial cells.
Methods: 10 patients with PHG underwent gastroscopy were randomly divided into two groups. After gastroscopy and specimen collection, one group was treated with propranolol hydrochloride (30mg/day), the other group was treated with propranolol hydrochloride (30mg/day) combined with teprenone capsule (150mg/day). Both groups were re-examined by gastroscopy one month after treatment. The specimens were collected again and the changes of gastric mucosal lesions were observed under endoscopy. In addition, 10 healthy volunteers who underwent upper gastrointestinal endoscopy were selected as healthy control group. The expression of in and ZO-1.
Results: The expression of tight junction protein Occludin and ZO-1 in gastric mucosa epithelial cells of PHG patients decreased. Before treatment, PHG patients were compared with healthy control group: 1. Gastroscopic manifestations: typical mosaic-like changes, snakeskin-like changes, scattered cherry red spots or diffuse hemorrhagic changes were observed in PHG patients. Appearance: There was obvious telangiectasia in the mucosa and submucosa of patients with PHG, but no obvious inflammation. 3. Immunohistochemical results of Occludin and ZO-1 protein: Occludin and ZO-1 protein in healthy control group were continuously distributed in the membrane of gastric mucosal epithelial cells in brown line, and drug trunk in patients with PHG. Compared with the healthy control group, Occludin protein and ZO-1 protein staining were significantly lighter, discontinuous, partially missing, and unevenly distributed. The mean optical density (OD) of Occludin was (0.03 (+) 0.01), (0.14 (+) 0.02), (P 0.001), and the OD value of ZO-1 were (0.03 (+) 0.01, (0.16 (+) 0.02), (P 0.001), respectively. Difference.
Propranolol hydrochloride can increase the expression of tight junction protein Occludin and ZO-1 in gastric mucosa epithelial cells of PHG patients. In the intervention group of propanolol hydrochloride alone, the gastroscopic manifestations of PHG patients after treatment were compared with those before treatment: (1) After treatment, the gastric mucosal lesions of PHG patients were improved. Immunohistochemical results of Occludin and ZO-1 proteins: After treatment, the expression of Occludin and ZO-1 proteins in gastric mucosal epithelial cells of the patients were deeper than before, and some of them were still missing, but the OD value of Occludin protein was lower than before, and the OD value of Occludin protein was (0.06 + 0.01), (0.03 + 0.01) respectively. .01), (P0.01), the OD values of ZO-1 protein were: (0.07 + 0.01), (0.03 + 0.01), (P0.01), there was statistical difference.
The expression of tight junction protein Occludin and ZO-1 in gastric mucosal epithelial cells of PHG patients was increased more significantly by the combination of propranolol hydrochloride and teprenolone than by propranolol hydrochloride alone. The following manifestations: The gastric mucosal lesions in the propranolol hydrochloride and teprenone group were significantly better than those in the propranolol hydrochloride alone. 2. Histomorphological findings: The telangiectasia in the propranolol hydrochloride and teprenone hydrochloride group was more obvious than that in the propranolol hydrochloride alone group. Immunohistochemical results of in protein and ZO-1 protein: After the combination of propranolol hydrochloride and teprenone treatment, the staining of Occludin protein and ZO-1 protein in gastric epithelial cells of PHG patients was deepened more significantly, and the deletion part was decreased more significantly. The OD value of Occludin protein was (0.11 (+) 0.01, (0.06 +) 0.01, (P 0.01), and the OD value of ZO-1 protein was (P 0.01). The values were: (0.12 + 0.02), (0.07 + 0.01), (P0.01), all of which were statistically different.
CONCLUSION: Decreased expression of tight junction proteins Occludin and ZO-1 in gastric epithelial cells of PHG patients may be an important cause of gastric mucosal barrier injury. Teprenone may protect gastric mucosal barrier by up-regulating the expression of Occludin and ZO-1.
【學位授予單位】：河北醫(yī)科大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R573

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