内皮保护剂治疗川崎病的作用及机制研究
发布时间：2019-06-03 19:49
【摘要】：目的： 探讨通过耳缘静脉重复注射异种动物蛋白诱导幼兔产生冠状动脉炎，验证其与川崎病冠状动脉病变的相似性，为研究川崎病提供实验模型。 方法： 20只4-6周龄的日本大耳幼兔随机分为模型组和对照组。两组分别给予10%牛血清白蛋白和生理盐水，第1天、第14天各重复注射一次。在首次注射后的第六周，两组幼兔分别行冠脉造影，并在造影结束后一小时之内显微镜下分离冠状动脉行组织病理学检测和超微结构的电镜观察，并留取肝脏、肾脏、肺脏及脾脏行组织病理学检查。 结果： (1)冠脉造影：仅模型组发现3只兔的冠状动脉出现不同程度的扩张或狭窄（3/10）；(2)组织病理学：模型组所有幼兔冠状动脉均出现内皮增厚、炎性细胞浸润，不连续的冠脉内皮；肺组织、肾脏组织呈现肉芽肿样改变主要由单核细胞组成，且肺组织病变部位小血管腔几乎完全被阻塞，部分肺泡塌陷，肺泡水肿不明显；肝脏组织出现点状坏死，单核细胞聚集；(3)透射电镜结果：模型组兔冠状动脉部分内弹力膜断裂，内皮不连续，伴有内皮细胞脱落和中层平滑肌细胞肿胀变性。 结论： 本研究证实牛血清白蛋白诱导产生的兔免疫性冠状动脉炎，类似川崎病冠状动脉炎改变，可作为模拟川崎病冠状动脉损伤的实验模型。 目的： 内皮细胞保护剂尚未作为川崎病的常规治疗手段，本研究拟在川崎病幼兔模型上，给予内皮保护剂辛伐他汀干预后，，观察其对川崎病的发病、病理改变及对内皮功能的影响，并探讨其中的机制，从而为从内皮细胞保护的角度防治川崎病提供初步的实验依据。 方法： 1.实验分组：将30只日本大耳幼兔随机分为模型组、辛伐他汀干预组、对照组三组。模型组及辛伐他汀干预组均予耳缘静脉注射牛血清白蛋白（10%，3ml/kg），对照组给予生理盐水（3ml/kg），第1天、第14天各重复注射一次。辛伐他汀干预组于第2次注射后连续三周每天予辛伐他汀干预（5mg/kg.d）灌胃；三组分别于第2次注射后第7、14、21天（即急性期、亚急性期、恢复期）分别采血一次。 2.检测方法： （1）评价建模成功的方法：①组织病理学检测各组幼兔的心肌及冠脉组织改变情况；②各组幼兔行冠状动脉造影检查； （2）ELISA法检测血浆内皮型一氧化氮合成酶（eNOS）的表达； （3）流式细胞仪计数法测定各组幼兔血浆内皮微颗粒（CD62E+EMPs、CD105+EMPs、CD144+/CD42b-EMPs）的表达水平； 结果： 1.组织病理学结果：模型组和辛伐他汀干预组均出现不同程度的炎症浸润。模型组可见血管内膜局部明显增厚，冠脉内皮不连续；而辛伐他汀干预组幼兔冠状动脉炎性细胞浸润较轻，内膜增厚不明显；对照组冠脉内皮光滑连续。 2. eNOS水平：辛伐他汀干预组eNOS表达水平低于模型组，P＜0.05，差异有统计学意义；辛伐他汀干预组eNOS表达水平与对照组相比，P＞0.05，差异无统计学意义。 3.辛伐他汀干预组KD幼兔急性期、亚急性期、恢复期血浆EMPs表达水平与模型组和对照组的比较： （1）辛伐他汀干预组及模型组三种表型的EMPs表达水平均高于对照组，P＜0.01，差异有显著的统计学意义； （2）辛伐他汀干预组与模型组进行比较：①CD62E+EMPs、CD105+EMPs水平比较：辛伐他汀干预组表达均低于模型组，P＜0.01，差异有显著的统计学意义；②CD144+/CD42b-EMPs：急性期辛伐他汀干预组表达低于模型组（P=0.001）P＜0.01，差异有显著的统计学意义；而亚急性期（P=0.199）恢复期（P=0.096）辛伐他汀干预组EMPs表达低于模型组，但差异无统计学意义。 4.模型组幼兔血浆EMPs表达水平： （1） CD62E+EMPs：急性期明显高于亚急性期、恢复期(P均＜0.05)差异有统计学意义；亚急性期与恢复期比较，P＞0.05，差异无统计学意义； （2） CD105+EMPs：急性期高于恢复期（P＜0.05）差异有统计学意义；急性期与亚急性期比较及亚急性期与恢复期比较，P均＞0.05，差异无统计学意义； （3） CD144+/CD42b-EMPs：恢复期表达高于急性期和亚急性期，亚急性期表达高于急性期，P均＜0.05，差异有统计学意义。 结论： 1.辛伐他汀可以降低内皮微颗粒的释放量、改善血管内皮功能障碍，推测辛伐他汀可能通过改善eNOS表达的稳定性，增加一氧化氮的生物利用度来改善内皮功能，eNOS参与了其发挥这一保护作用的机制，为KD血管内皮的保护提供依据。 2. KD兔模型组EMPs持续增高或降低缓慢提示KD恢复期内皮损伤持续存在；EMPs水平的监测尤其是CD144+/CD42b-EMPs可用于KD长期预后评估。
[Abstract]:Purpose: To study the similarity of coronary artery disease with Kawasaki disease by repeated injection of different animal protein through the ear-margin vein, and to provide experimental model for the study of Kawasaki disease. Type. Methods: Twenty four-six-week-old Japanese young rabbits were randomly divided into two groups. The two groups were given 10% bovine serum albumin and normal saline, day 1, day 14 each In the sixth week after the first injection, the two groups of young rabbits underwent coronary angiography respectively, and the pathological examination and the ultrastructure of the coronary artery were observed under the microscope at an hour after the end of the contrast, and the liver, the kidney, the lung and the spleen were collected. pathology Results: (1) Coronary angiography: only the model group found that the coronary arteries of 3 rabbits had different degrees of expansion or stenosis (3/10); (2) Histopathology: All the young rabbits in the model group had internal skin thickening, inflammatory cell infiltration, and the small blood vessel cavity in the pathological part of the lung tissue is almost completely blocked, the partial alveoli collapse and the alveolar edema is not obvious, and the liver tissue has the point necrosis and the monocyte aggregation; (3) Transmission electron microscope (TEM) results: The elastic membrane in the coronary part of the model group is broken, the endothelium is not continuous, and the endothelial cells fall off and the middle layer smooth muscle Conclusion: This study confirmed that bovine serum albumin induced an immune coronary arteritis, similar to that of Kawasaki disease, and can be used as an analog. Saaki's disease Objective: The protective agent of endothelial cell has not been used as a routine treatment for Kawasaki disease. The effects of the endothelial function and the mechanism of the mechanism were discussed, so as to protect the endothelial cells from the endothelial cells. the angle of the A preliminary experimental basis for the prevention and treatment of Kawasaki disease. Method:1. Experimental grouping:30 Japanese large-ear young rabbits The serum albumin (10%,3 ml/ kg) of bovine serum albumin (10%,3 ml/ kg) was given to the ear margin in the model group and the simvastatin intervention group, and the control group was given normal saline (3 m (l/ kg), Day 1, and Day 14. Simvastatin intervention (5 mg/ kg. d) for three weeks after the second injection, and 7,14,21 days after the second injection, respectively. (i.e. acute phase, suba 2. Test method: (1) The method of evaluating the success of modeling: the heart of each group of young rabbits is detected by histopathology Changes of muscle and coronary tissue; coronary angiography of young rabbits in each group; (2) EL The expression of endothelial nitric oxide synthase (eNOS) in plasma was detected by ISA method. (3) The plasma endothelial microgranules (CD62E + EMPs, CD105) were determined by flow cytometry. + EM Ps, CD144 +/ CD42b-EMPs); Results: 1. Histopathological results: There was a different degree of inflammatory infiltrates in both the model group and the simvastatin intervention group. In the control group, the expression level of eNOS was lower than that of the model group (P <0.05). Compared with the control group, the expression level of eNOS in the statin intervention group was significantly higher than that of the control group (P> 0.05, and the difference was not significant.3. Simvastatin intervention group The expression levels of EMPs in the acute, subacute and convalescent phases of KD were compared with the model group and the control group: 1) Simvastatin intervention group and model The expression of EMPs in three groups was higher than that in the control group (P <0.01). (2) Simvastatin intervention group was compared with the model group: the expression of CD62E + EMPs, CD105 + EMPs was lower than that of the control group. Compared with the model group (P = 0.001), the expression of CD144 +/ CD42b-EMPs was significantly lower than that of the model group (P = 0.001). ) Recovery period (P = 0.096) Simvastatin Dry The expression of EMPs in the pregroup was lower than that of the model group, but the difference was not significant.4. The expression level of EMPs in the model group was: (1) CD62E + EMPs: the acute stage was significantly higher than that of the subacute stage. The difference of the period (P <0.05) is in series. (2) CD105 + EMPs: The acute phase was higher than the recovery period (P <0.05). There was a significant difference in the difference between the acute and the subacute phase and the recovery period compared with the recovery period, all of which were more than 0.0. 5. No statistical significance; (3) CD144 +/ CD42b- EMP "s: The expression of the recovery period is higher than that in the acute and subacute stages, and the expression of the subacute stage is higher than that of the acute stage, P <0.05, the difference is of statistical significance. Conclusion:1. Simvastatin can reduce the release of the endothelial micro-particles, To improve vascular endothelial dysfunction, it is suggested that simvastatin may increase the stability of eNOS expression by increasing the stability of eNOS expression. The biological utilization of nitric oxide improves endothelial function, and the eNOS is involved in the mechanism of this protective effect and provides a basis for the protection of KD vascular endothelium.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.4
本文编号：2492180
[Abstract]:Purpose: To study the similarity of coronary artery disease with Kawasaki disease by repeated injection of different animal protein through the ear-margin vein, and to provide experimental model for the study of Kawasaki disease. Type. Methods: Twenty four-six-week-old Japanese young rabbits were randomly divided into two groups. The two groups were given 10% bovine serum albumin and normal saline, day 1, day 14 each In the sixth week after the first injection, the two groups of young rabbits underwent coronary angiography respectively, and the pathological examination and the ultrastructure of the coronary artery were observed under the microscope at an hour after the end of the contrast, and the liver, the kidney, the lung and the spleen were collected. pathology Results: (1) Coronary angiography: only the model group found that the coronary arteries of 3 rabbits had different degrees of expansion or stenosis (3/10); (2) Histopathology: All the young rabbits in the model group had internal skin thickening, inflammatory cell infiltration, and the small blood vessel cavity in the pathological part of the lung tissue is almost completely blocked, the partial alveoli collapse and the alveolar edema is not obvious, and the liver tissue has the point necrosis and the monocyte aggregation; (3) Transmission electron microscope (TEM) results: The elastic membrane in the coronary part of the model group is broken, the endothelium is not continuous, and the endothelial cells fall off and the middle layer smooth muscle Conclusion: This study confirmed that bovine serum albumin induced an immune coronary arteritis, similar to that of Kawasaki disease, and can be used as an analog. Saaki's disease Objective: The protective agent of endothelial cell has not been used as a routine treatment for Kawasaki disease. The effects of the endothelial function and the mechanism of the mechanism were discussed, so as to protect the endothelial cells from the endothelial cells. the angle of the A preliminary experimental basis for the prevention and treatment of Kawasaki disease. Method:1. Experimental grouping:30 Japanese large-ear young rabbits The serum albumin (10%,3 ml/ kg) of bovine serum albumin (10%,3 ml/ kg) was given to the ear margin in the model group and the simvastatin intervention group, and the control group was given normal saline (3 m (l/ kg), Day 1, and Day 14. Simvastatin intervention (5 mg/ kg. d) for three weeks after the second injection, and 7,14,21 days after the second injection, respectively. (i.e. acute phase, suba 2. Test method: (1) The method of evaluating the success of modeling: the heart of each group of young rabbits is detected by histopathology Changes of muscle and coronary tissue; coronary angiography of young rabbits in each group; (2) EL The expression of endothelial nitric oxide synthase (eNOS) in plasma was detected by ISA method. (3) The plasma endothelial microgranules (CD62E + EMPs, CD105) were determined by flow cytometry. + EM Ps, CD144 +/ CD42b-EMPs); Results: 1. Histopathological results: There was a different degree of inflammatory infiltrates in both the model group and the simvastatin intervention group. In the control group, the expression level of eNOS was lower than that of the model group (P <0.05). Compared with the control group, the expression level of eNOS in the statin intervention group was significantly higher than that of the control group (P> 0.05, and the difference was not significant.3. Simvastatin intervention group The expression levels of EMPs in the acute, subacute and convalescent phases of KD were compared with the model group and the control group: 1) Simvastatin intervention group and model The expression of EMPs in three groups was higher than that in the control group (P <0.01). (2) Simvastatin intervention group was compared with the model group: the expression of CD62E + EMPs, CD105 + EMPs was lower than that of the control group. Compared with the model group (P = 0.001), the expression of CD144 +/ CD42b-EMPs was significantly lower than that of the model group (P = 0.001). ) Recovery period (P = 0.096) Simvastatin Dry The expression of EMPs in the pregroup was lower than that of the model group, but the difference was not significant.4. The expression level of EMPs in the model group was: (1) CD62E + EMPs: the acute stage was significantly higher than that of the subacute stage. The difference of the period (P <0.05) is in series. (2) CD105 + EMPs: The acute phase was higher than the recovery period (P <0.05). There was a significant difference in the difference between the acute and the subacute phase and the recovery period compared with the recovery period, all of which were more than 0.0. 5. No statistical significance; (3) CD144 +/ CD42b- EMP "s: The expression of the recovery period is higher than that in the acute and subacute stages, and the expression of the subacute stage is higher than that of the acute stage, P <0.05, the difference is of statistical significance. Conclusion:1. Simvastatin can reduce the release of the endothelial micro-particles, To improve vascular endothelial dysfunction, it is suggested that simvastatin may increase the stability of eNOS expression by increasing the stability of eNOS expression. The biological utilization of nitric oxide improves endothelial function, and the eNOS is involved in the mechanism of this protective effect and provides a basis for the protection of KD vascular endothelium.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R725.4
【参考文献】
相关期刊论文 前2条
1 于明华;小儿川崎病的研究现状和展望[J];广东医学;2002年07期
2 韦卫中,陈绍军,王宏伟;免疫性血管炎致冠状动脉扩张的实验研究[J];中华儿科杂志;2003年03期
本文编号：2492180
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