【摘要】：目的： 探討通過耳緣靜脈重復注射異種動物蛋白誘導幼兔產生冠狀動脈炎，驗證其與川崎病冠狀動脈病變的相似性，為研究川崎病提供實驗模型。 方法： 20只4-6周齡的日本大耳幼兔隨機分為模型組和對照組。兩組分別給予10%牛血清白蛋白和生理鹽水，第1天、第14天各重復注射一次。在首次注射后的第六周，兩組幼兔分別行冠脈造影，并在造影結束后一小時之內顯微鏡下分離冠狀動脈行組織病理學檢測和超微結構的電鏡觀察，并留取肝臟、腎臟、肺臟及脾臟行組織病理學檢查。 結果： (1)冠脈造影：僅模型組發(fā)現3只兔的冠狀動脈出現不同程度的擴張或狹窄（3/10）；(2)組織病理學：模型組所有幼兔冠狀動脈均出現內皮增厚、炎性細胞浸潤，不連續(xù)的冠脈內皮；肺組織、腎臟組織呈現肉芽腫樣改變主要由單核細胞組成，且肺組織病變部位小血管腔幾乎完全被阻塞，部分肺泡塌陷，肺泡水腫不明顯；肝臟組織出現點狀壞死，單核細胞聚集；(3)透射電鏡結果：模型組兔冠狀動脈部分內彈力膜斷裂，內皮不連續(xù)，伴有內皮細胞脫落和中層平滑肌細胞腫脹變性。 結論： 本研究證實牛血清白蛋白誘導產生的兔免疫性冠狀動脈炎，類似川崎病冠狀動脈炎改變，可作為模擬川崎病冠狀動脈損傷的實驗模型。 目的： 內皮細胞保護劑尚未作為川崎病的常規(guī)治療手段，本研究擬在川崎病幼兔模型上，給予內皮保護劑辛伐他汀干預后，，觀察其對川崎病的發(fā)病、病理改變及對內皮功能的影響，并探討其中的機制，從而為從內皮細胞保護的角度防治川崎病提供初步的實驗依據。 方法： 1.實驗分組：將30只日本大耳幼兔隨機分為模型組、辛伐他汀干預組、對照組三組。模型組及辛伐他汀干預組均予耳緣靜脈注射牛血清白蛋白（10%，3ml/kg），對照組給予生理鹽水（3ml/kg），第1天、第14天各重復注射一次。辛伐他汀干預組于第2次注射后連續(xù)三周每天予辛伐他汀干預（5mg/kg.d）灌胃；三組分別于第2次注射后第7、14、21天（即急性期、亞急性期、恢復期）分別采血一次。 2.檢測方法： （1）評價建模成功的方法：①組織病理學檢測各組幼兔的心肌及冠脈組織改變情況；②各組幼兔行冠狀動脈造影檢查； （2）ELISA法檢測血漿內皮型一氧化氮合成酶（eNOS）的表達； （3）流式細胞儀計數法測定各組幼兔血漿內皮微顆粒（CD62E+EMPs、CD105+EMPs、CD144+/CD42b-EMPs）的表達水平； 結果： 1.組織病理學結果：模型組和辛伐他汀干預組均出現不同程度的炎癥浸潤。模型組可見血管內膜局部明顯增厚，冠脈內皮不連續(xù)；而辛伐他汀干預組幼兔冠狀動脈炎性細胞浸潤較輕，內膜增厚不明顯；對照組冠脈內皮光滑連續(xù)。 2. eNOS水平：辛伐他汀干預組eNOS表達水平低于模型組，P＜0.05，差異有統(tǒng)計學意義；辛伐他汀干預組eNOS表達水平與對照組相比，P＞0.05，差異無統(tǒng)計學意義。 3.辛伐他汀干預組KD幼兔急性期、亞急性期、恢復期血漿EMPs表達水平與模型組和對照組的比較： （1）辛伐他汀干預組及模型組三種表型的EMPs表達水平均高于對照組，P＜0.01，差異有顯著的統(tǒng)計學意義； （2）辛伐他汀干預組與模型組進行比較：①CD62E+EMPs、CD105+EMPs水平比較：辛伐他汀干預組表達均低于模型組，P＜0.01，差異有顯著的統(tǒng)計學意義；②CD144+/CD42b-EMPs：急性期辛伐他汀干預組表達低于模型組（P=0.001）P＜0.01，差異有顯著的統(tǒng)計學意義；而亞急性期（P=0.199）恢復期（P=0.096）辛伐他汀干預組EMPs表達低于模型組，但差異無統(tǒng)計學意義。 4.模型組幼兔血漿EMPs表達水平： （1） CD62E+EMPs：急性期明顯高于亞急性期、恢復期(P均＜0.05)差異有統(tǒng)計學意義；亞急性期與恢復期比較，P＞0.05，差異無統(tǒng)計學意義； （2） CD105+EMPs：急性期高于恢復期（P＜0.05）差異有統(tǒng)計學意義；急性期與亞急性期比較及亞急性期與恢復期比較，P均＞0.05，差異無統(tǒng)計學意義； （3） CD144+/CD42b-EMPs：恢復期表達高于急性期和亞急性期，亞急性期表達高于急性期，P均＜0.05，差異有統(tǒng)計學意義。 結論： 1.辛伐他汀可以降低內皮微顆粒的釋放量、改善血管內皮功能障礙，推測辛伐他汀可能通過改善eNOS表達的穩(wěn)定性，增加一氧化氮的生物利用度來改善內皮功能，eNOS參與了其發(fā)揮這一保護作用的機制，為KD血管內皮的保護提供依據。 2. KD兔模型組EMPs持續(xù)增高或降低緩慢提示KD恢復期內皮損傷持續(xù)存在；EMPs水平的監(jiān)測尤其是CD144+/CD42b-EMPs可用于KD長期預后評估。
[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.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2013
【分類號】：R725.4

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