【摘要】：目的 應激性胃潰瘍出血是急性腦血管病等諸多重大顱腦損傷最常見的并發(fā)癥之一,一旦發(fā)生很難控制,死亡率極高。因此,胃黏膜損傷與保護是永恒的主題。急性腦血管疾病所致胃腸損傷涉及眾多的病理生理過程,關于其發(fā)生機制雖然做了不少研究,但其確切機制仍不十分清楚。本文通過比較大鼠腦缺血再灌注狀態(tài)下的胃黏膜與胃局部缺血再灌注胃黏膜的組織學與組織化學變化的異同；激光多普勒血流儀檢測大鼠腦缺血再灌注前后胃黏膜血流變化；結合血管活性肽—降鈣素基因相關肽對胃黏膜損傷作用實驗觀察,綜合探討胃血流灌注量與急性腦血管病所致胃黏膜損傷的關系。 方法 1.健康Wistar大鼠,體重280～32g,隨機分為4組,即腦缺血再灌注(MCAO)組、腦缺血假手術(MCAO-sham)組、胃缺血再灌注(GI-R)組和胃缺血假手術(GI-R-sham)組,每組12只大鼠。采用線栓法制作大鼠局灶性腦缺血再灌注模型。按Zea longa等5分制方法對其進行評分,得分在1-3分者被認為模型制作成功,納入實驗。腦缺血假手術組只模擬手術,不阻塞大腦中動脈。MCAO組于腦缺血1h后實現(xiàn)再灌注。按Wada等方法,制造大鼠胃缺血再灌注型模型。動物開腹,仔細分離腹腔動脈及其周圍組織,夾閉腹腔動脈30min,去除動脈夾恢復血流。假手術(GI-R-sham組)僅分離腹腔動脈,不進行夾閉。四組均于術后48h取材：參考Guth等方法觀察胃黏膜損傷指數(shù)；光鏡觀察腦缺血再灌注后胃黏膜的形態(tài)學改變；免疫組織化學方法檢測胃黏膜組織胃泌素(gastrin, Gas)、生長抑素(somatostatin, SST),—抗為兔抗SST抗體和兔抗Gas抗體,每項指標檢測6只大鼠。所得數(shù)據(jù)行單因素方差分析,兩組間比較用9檢驗。 2.健康Wistar大鼠,體重280-320g,隨機分為5組,即假手術(Sham)組,生理鹽水(NS)組,低劑量CGRP組,中劑量CGRP組,高劑量CGRP組,每組10只。腦梗死前的基礎胃黏膜血流測試：3.5%水合氯醛麻醉不同組大鼠,頸部和腹部備皮,仰臥位固定于手術臺上。沿腹白線打開腹腔,暴露大鼠胃部。用20m1注射器針頭從胃底插入胃腔,注意不要損傷其它部位的胃黏膜,將激光多普勒纖維探頭從針孔中穿進胃腔,每隔45秒按照胃小彎、胃大彎、胃前壁、胃后壁的順序分別測量這4處的胃血流,循環(huán)檢測三次,取其平均值代表腦梗死前的基礎胃黏膜血流。腦梗死后的胃黏膜血流測試：將針頭從胃腔中拔出,浸泡入生理鹽水中。用溫生理鹽水浸泡過的紗布將暴露的胃部包裹,用結扎線將胃底的針孔輕輕結扎,無胃液滲出即可,防止其污染腹腔。用生理鹽水紗布覆蓋腹部創(chuàng)口。建立大鼠局灶性腦缺血再灌注模型,方法同上。魚線栓塞大腦中動脈后,即將針頭重新從針孔中插入胃腔,按照同樣的方法連續(xù)檢測胃黏膜血流量。1小時后將魚線拔出少許,腦血流恢復再灌注,同時給予NS(1ml/100g), CGRP:低劑量組(1.5ug/ml,1ml/100g),中劑量組(3ug/ml,1ml/100g),高劑量組(6ug/ml,1ml/100g)。繼續(xù)檢測胃血流30分鐘。免疫組織化學方法檢測CD31在胃黏膜中的表達,一抗為兔抗CD31抗體。所得數(shù)據(jù)行單因素方差分析,兩組間比較用9檢驗。結果 1.腦缺血再灌注大鼠①體視顯微鏡下NS組胃黏膜損傷嚴重,見彌漫性水腫及點片狀出血、糜爛。CGRP組胃黏膜損傷指數(shù)小于NS組(P＜0.05)。②HE染色見NS組黏膜上皮細胞明顯受損,有壞死、脫落,并可見中性粒細胞,單核細胞等炎性細胞浸潤,腺體排列紊亂。CGRP組病變明顯減輕,僅見胃黏膜部分脫落,腺體排列稍紊亂,較少見胃黏膜出血及炎癥細胞浸潤。③CGRP組胃竇部黏膜胃泌素的表達低于NS組(P0.01),SST的表達高于NS組(P＜0.01)。 2.激光多普勒血流儀檢測結果顯示：大鼠腦梗死后胃黏膜血流量下降,給予外源性CGRP胃黏膜血流量有一定程度上升。中、高劑量組高于NS組(P0.05)；低劑量組低于中劑量組(P＜0.05)}；高劑量組高于中劑量組(P0.05)。免疫組織化學檢測CD31在胃黏膜上的表達量結果顯示：CGRP組和sham組大鼠胃黏膜CD31表達量高于NS組(P0.05)。緒論 胃黏膜血流灌注降低是大鼠急性腦缺血再灌注致胃黏膜損傷的首要因素；微循環(huán)障礙是胃黏膜應激損傷的中心環(huán)節(jié)；CGRP保護缺血性胃黏膜損傷的重要機制之一是增加胃黏膜血流灌注,改善微循環(huán)。
[Abstract]:Purpose Stress gastric ulcer is one of the most common complications such as acute cerebrovascular disease. Once it is difficult to control, the mortality rate is extremely high. High. Therefore, gastric mucosal injury and protection are the eternal main The gastrointestinal injury caused by acute cerebrovascular disease involves many pathophysiological processes. Although many studies have been done on the mechanism of the occurrence of the gastrointestinal injury, the exact mechanism is still not very clear. Objective: To compare the histological and tissue chemical changes of the gastric mucosa and the local ischemia-reperfusion gastric mucosa in rats after cerebral ischemia-reperfusion in rats. The blood flow of the gastric mucosa before and after the cerebral ischemia-reperfusion in rats was detected by the laser Doppler flow meter. Objective: To study the effect of vasoactive peptide and calcitonin gene-related peptide on gastric mucosal injury, and to study the effect of gastric perfusion on gastric mucosal injury caused by acute cerebrovascular disease. Department. Methods 1. Healthy Wistar rats, weighing 280-32g, were randomly divided into 4 groups, namely, cerebral ischemia-reperfusion (MCAO) group, cerebral ischemia-reperfusion (MCAO-sham) group, gastric ischemia-reperfusion (GI-R) group and gastric ischemia-sham operation (GI-R-sham) group, each group 12 rats. Rats with focal cerebral ischemia were prepared by the method of line-bolt. The model was reperfused. The score was scored according to the five-point method, such as Zea lona, and the score was 1-3. The model was considered to be successful. It was included in the experimental group. The operation group was only simulated for operation and not blocked. The middle cerebral artery. The MCAO group was after 1 h of cerebral ischemia. The method of Wada and the like can be used for manufacturing the rat gastric ischemia Model of perfusion type. The animal was open to open, the celiac artery and its surrounding tissues were carefully separated, the celiac artery was closed for 30min, and the artery was removed. the clip recovered the blood flow. the sham operation (gi-r-sham group) only separates the celiac artery, The gastric mucosal injury index was observed by the method of Guth et al. The morphological changes of the gastric mucosa after cerebral ischemia and reperfusion were observed by light microscope, and the gastrin (Gas) and somatostatin (somatosstatin) in the gastric mucosa were detected by the immunohistochemical method. (SST), anti-SST antibody and rabbit anti-Gas antibody for rabbit anti-SST antibody and rabbit anti-Gas antibody Six rats were measured. The resulting data line was one-factor analysis of variance, the ratio between the two groups Two healthy Wistar rats were randomly divided into 5 groups, namely, sham operation (Sham) group, normal saline (NS) group, low-dose CGRP group, middle-dose CGRP group, high-dose CGRP group, The basic gastric mucosal blood flow test before cerebral infarction: 3. 5% hydrochloral anesthesia in different groups of rats, the neck and the abdomen, and the supine position To be fixed on the operating table. Open the abdominal cavity along the white line of the abdomen. The stomach of the rat was exposed. The stomach cavity was inserted from the bottom of the stomach with a 20m1 syringe needle. It was noted that the gastric mucosa of the other parts should not be damaged. The laser Doppler fiber probe was inserted into the stomach cavity from the needle hole, and measured in the order of the stomach small curve, the large intestine, the front wall of the stomach and the back wall of the stomach, respectively, every 45 seconds. The blood flow of the stomach at this 4 was measured three times, and the mean value was taken to represent the pre-cerebral infarction. Basic gastric mucosal blood flow. Gastric mucosal blood flow test following cerebral infarction: Pull the needle out of the stomach cavity and soak soaking in physiological saline, wrapping the exposed stomach with the gauze soaked with warm saline, gently ligature the needle hole of the stomach bottom with the ligature wire, and not exudation of the gastric juice; to prevent it from contaminating the abdominal cavity. Cloth covering the abdominal wound. Establishment of focal cerebral ischemia-reperfusion in rats The model and method were the same as the above. After the fish line was embolized into the middle cerebral artery, the needle was re-inserted into the stomach cavity from the needle hole, and the blood flow of the gastric mucosa was continuously detected by the same method. After 1 hour, the fish line was pulled out a little, the cerebral blood flow was recovered and reperfused, and the NS (1ml/ 100g), the CGRP, the low-dose group (1. 5ug/ ml, 1ml/ 100g) and the middle-dose group were given. (3ug/ ml, 1ml/ 100g), high dose group (6ug/ ml, 1m l/ 100g). Continue to detect Gastric blood flow for 30 min. Immunohistochemistry was used to detect the expression of CD31 in the gastric mucosa. Anti-CD31 antibody. One-factor analysis of variance for the resulting data line, two groups interspecific ratio 9. Results 1. The damage of the gastric mucosa of NS group in the rats with cerebral ischemia and reperfusion was severe, and diffuse edema was found. and the damage index of the gastric mucosa of the CGRP group was less than that of NS. In the group (P <0.05), the mucosa epithelial cells of the NS group were obviously damaged, the necrosis and the shedding were observed in the HHE staining, and the inflammatory cells such as neutrophils and monocytes were observed. In the CGRP group, the lesions of the CGRP group were significantly reduced, only the partial shedding of the gastric mucosa and the slight disturbance of the gland, and the less common gastric mucosa. The expression of gastrin in the GCGRP group was lower than that in NS group (P0.01), and the expression of SST was higher than that in NS group (P0.01). The results of the laser Doppler flow cytometry showed that the blood flow of the gastric mucosa decreased after the cerebral infarction in the rats, and the exogenous CGRP was given to the stomach. The high-dose group was higher than that in NS group (P <0.05), and the high-dose group was higher than that of middle-dose group (P <0.05); the high-dose group was higher than that of the middle-dose group (P <0.05). The expression of CD31 in the gastric mucosa was detected by immunohistochemistry in the middle dose group (P0.05). The expression of CD31 in the gastric mucosa of the CGRP group and the sham group was high. N The decrease of blood flow perfusion in gastric mucosa is the primary factor in gastric mucosal injury induced by acute cerebral ischemia-reperfusion in rats. Microcirculation disorder is the central part of gastric mucosal stress injury. One of the important mechanisms of CGRP to protect ischemic gastric mucosa is to increase
【學位授予單位】：濱州醫(yī)學院
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R743.3

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