胃血流灌注量与大鼠急性脑梗死致胃黏膜应激性损伤关系的研究
发布时间：2019-02-21 13:06
【摘要】：目的 应激性胃溃疡出血是急性脑血管病等诸多重大颅脑损伤最常见的并发症之一,一旦发生很难控制,死亡率极高。因此,胃黏膜损伤与保护是永恒的主题。急性脑血管疾病所致胃肠损伤涉及众多的病理生理过程,关于其发生机制虽然做了不少研究,但其确切机制仍不十分清楚。本文通过比较大鼠脑缺血再灌注状态下的胃黏膜与胃局部缺血再灌注胃黏膜的组织学与组织化学变化的异同；激光多普勒血流仪检测大鼠脑缺血再灌注前后胃黏膜血流变化；结合血管活性肽—降钙素基因相关肽对胃黏膜损伤作用实验观察,综合探讨胃血流灌注量与急性脑血管病所致胃黏膜损伤的关系。 方法 1.健康Wistar大鼠,体重280～32g,随机分为4组,即脑缺血再灌注(MCAO)组、脑缺血假手术(MCAO-sham)组、胃缺血再灌注(GI-R)组和胃缺血假手术(GI-R-sham)组,每组12只大鼠。采用线栓法制作大鼠局灶性脑缺血再灌注模型。按Zea longa等5分制方法对其进行评分,得分在1-3分者被认为模型制作成功,纳入实验。脑缺血假手术组只模拟手术,不阻塞大脑中动脉。MCAO组于脑缺血1h后实现再灌注。按Wada等方法,制造大鼠胃缺血再灌注型模型。动物开腹,仔细分离腹腔动脉及其周围组织,夹闭腹腔动脉30min,去除动脉夹恢复血流。假手术(GI-R-sham组)仅分离腹腔动脉,不进行夹闭。四组均于术后48h取材：参考Guth等方法观察胃黏膜损伤指数；光镜观察脑缺血再灌注后胃黏膜的形态学改变；免疫组织化学方法检测胃黏膜组织胃泌素(gastrin, Gas)、生长抑素(somatostatin, SST),—抗为兔抗SST抗体和兔抗Gas抗体,每项指标检测6只大鼠。所得数据行单因素方差分析,两组间比较用9检验。 2.健康Wistar大鼠,体重280-320g,随机分为5组,即假手术(Sham)组,生理盐水(NS)组,低剂量CGRP组,中剂量CGRP组,高剂量CGRP组,每组10只。脑梗死前的基础胃黏膜血流测试：3.5%水合氯醛麻醉不同组大鼠,颈部和腹部备皮,仰卧位固定于手术台上。沿腹白线打开腹腔,暴露大鼠胃部。用20m1注射器针头从胃底插入胃腔,注意不要损伤其它部位的胃黏膜,将激光多普勒纤维探头从针孔中穿进胃腔,每隔45秒按照胃小弯、胃大弯、胃前壁、胃后壁的顺序分别测量这4处的胃血流,循环检测三次,取其平均值代表脑梗死前的基础胃黏膜血流。脑梗死后的胃黏膜血流测试：将针头从胃腔中拔出,浸泡入生理盐水中。用温生理盐水浸泡过的纱布将暴露的胃部包裹,用结扎线将胃底的针孔轻轻结扎,无胃液渗出即可,防止其污染腹腔。用生理盐水纱布覆盖腹部创口。建立大鼠局灶性脑缺血再灌注模型,方法同上。鱼线栓塞大脑中动脉后,即将针头重新从针孔中插入胃腔,按照同样的方法连续检测胃黏膜血流量。1小时后将鱼线拔出少许,脑血流恢复再灌注,同时给予NS(1ml/100g), CGRP:低剂量组(1.5ug/ml,1ml/100g),中剂量组(3ug/ml,1ml/100g),高剂量组(6ug/ml,1ml/100g)。继续检测胃血流30分钟。免疫组织化学方法检测CD31在胃黏膜中的表达,一抗为兔抗CD31抗体。所得数据行单因素方差分析,两组间比较用9检验。结果 1.脑缺血再灌注大鼠①体视显微镜下NS组胃黏膜损伤严重,见弥漫性水肿及点片状出血、糜烂。CGRP组胃黏膜损伤指数小于NS组(P＜0.05)。②HE染色见NS组黏膜上皮细胞明显受损,有坏死、脱落,并可见中性粒细胞,单核细胞等炎性细胞浸润,腺体排列紊乱。CGRP组病变明显减轻,仅见胃黏膜部分脱落,腺体排列稍紊乱,较少见胃黏膜出血及炎症细胞浸润。③CGRP组胃窦部黏膜胃泌素的表达低于NS组(P0.01),SST的表达高于NS组(P＜0.01)。 2.激光多普勒血流仪检测结果显示：大鼠脑梗死后胃黏膜血流量下降,给予外源性CGRP胃黏膜血流量有一定程度上升。中、高剂量组高于NS组(P0.05)；低剂量组低于中剂量组(P＜0.05)}；高剂量组高于中剂量组(P0.05)。免疫组织化学检测CD31在胃黏膜上的表达量结果显示：CGRP组和sham组大鼠胃黏膜CD31表达量高于NS组(P0.05)。绪论 胃黏膜血流灌注降低是大鼠急性脑缺血再灌注致胃黏膜损伤的首要因素；微循环障碍是胃黏膜应激损伤的中心环节；CGRP保护缺血性胃黏膜损伤的重要机制之一是增加胃黏膜血流灌注,改善微循环。
[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
【学位授予单位】：滨州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R743.3
本文编号：2427530
[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
【学位授予单位】：滨州医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R743.3
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