本文關(guān)鍵詞：肺血少型先心病幼豬動物模型的建立及肺血改變對未成熟肺血管形態(tài)發(fā)育和功能的影響　出處：《中國協(xié)和醫(yī)科大學(xué)》2008年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 先天性心臟病 紫紺 動物模型 豬 形態(tài)定量分析 肺動脈 發(fā)育不良 先天性心臟病 紫紺 肺動脈 發(fā)育不良 基質(zhì)金屬蛋白酶 組織基質(zhì)金屬蛋白酶抑制劑 血管內(nèi)皮細(xì)胞生長因子 Ⅰ型和Ⅲ型膠原 肺發(fā)育

【摘要】： 第一部分.介入和外科技術(shù)相結(jié)合建立紫紺型肺血少先心病幼豬動物模型 【目的】.采用一種新穎的介入和外科相結(jié)合的Hybrid方法建立紫紺型肺血少先心病動物模型。 【方法】.采用生后1～2月的幼豬,體重8～12kg,分為兩組,每組8只。a)正常對照組(C-group,n=8):即假手術(shù)組,只開胸?zé)o手術(shù)處理;b)肺血減少組(D-group,n=8):右前外側(cè)第三肋間開胸,經(jīng)右心房表面送入擴張器行人工房間隔造口術(shù);再經(jīng)左前外側(cè)第三肋間開胸,將主肺動脈套帶環(huán)縮。所有動物正常飼養(yǎng)2個月。然后正中開胸,測量跨肺動脈環(huán)縮處的壓差;分別抽外周動脈、右心房、肺靜脈、左心房血液行血氣分析,計算Qp/Qs;抽外周靜脈血測量血球壓積(HCT)和血紅蛋白濃度(HB)。 【結(jié)果】.對照組8只均存活,平均體重增加到23.0kg。肺血減少組存活6只,平均體重增加到20.6kg。肺血減少組的動脈血氧分壓(PaO_2)、動脈血氧飽和度(SaO_2)、HCT和HB分別為52.9 mmHg、85.6%、49.8%和16.6g/dl。對照組的PaO_2、SaO_2、HCT和HB則分別為118.0 mmHg、98.0%、37.9%和12.2g/dl。二組間對應(yīng)指標(biāo)相比均有顯著性差異(P＜0.001)。利用四點血氣值和公式計算所得,肺血減少組的Qp/Qs為0.54:1。 【結(jié)論】.應(yīng)用介入和外科相結(jié)合的Hybrid方法,通過人工房間隔造口和肺動脈環(huán)縮,成功建立了一種新穎的紫紺型肺血少先天性心臟病動物模型,為研究紫紺型肺血少先心病的臨床病理和生理提供了一種有用的工具。 第二部分.肺血減少肺動脈低壓時肺組織的病理形態(tài)觀察和分析 【目的】.利用我們已建立的紫紺型肺血少動物模型,對肺血減少時未成熟肺小血管進行病理形態(tài)觀察和形態(tài)半定量分析,研究肺血減少時未成熟肺小血管的發(fā)育特點。 【方法】.健康幼豬16只,分為紫紺肺血減少組(D組,n=8)和對照組(C組,n=8)。首先利用第一部分描述的方法建立紫紺型肺血少先心病動物模型。兩組動物正常飼養(yǎng)2個月,麻醉和正中開胸,按照動物活檢取材步驟,分別取雙側(cè)上、中、下肺葉組織各一塊,4%的多聚甲醛充分固定后制片,行H-E和彈性蛋白+Van Gieson染色。光鏡和透射電鏡觀察兩組動物肺小血管和肺實質(zhì)的病理形態(tài)特征;利用顯微鏡-微機圖像處理系統(tǒng),形態(tài)半定量測量分析肺小動脈平均中膜厚度(MT)及中膜厚度百分比(MT%)、平均中膜面積(MS)及中膜面積百分比(MS%);計數(shù)單位面積肺小動脈數(shù)目(APSC)和單位面積肺泡數(shù)量(MAN);計算同一張切片所有泡內(nèi)肺動脈(IAPA)中肌型動脈(CMA)、部分肌型動脈(PMA)和無肌型動脈(NMA)的比例。 【結(jié)果】.對照組(C組)8只均存活;肺血減少組(D組)存活6只,D組成功建立紫紺型肺血少先心病動物模型。同對照C組動物相比,光鏡觀察下D組肺小動脈管腔普遍擴大,形態(tài)不規(guī)則,管壁厚薄不均,部分彈力纖維層中斷、缺失,肺小動脈中層退化和發(fā)育不良,部分肌型動脈和無肌型動脈的比例較對照組高,而肌型動脈的比例卻相對較低(P＜0.001)。D組肺小動脈MT絕對值、MT%及MS、MS%均顯著小于C組(均為P＜0.001)。D組單位面積肺小動脈數(shù)(APSC)較C組少(P＜0.01),而肺泡數(shù)與C組無顯著差異(P＞0.05)。電鏡觀察顯示肺血減少組肺小血管內(nèi)皮細(xì)胞胞質(zhì)腫脹,基底膜變薄;彈力纖維排列不整齊,有的區(qū)域萎縮變薄或中斷;血管中層平滑肌輕度萎縮;細(xì)胞器減少,線粒體空泡化:部分肺泡Ⅱ型上皮細(xì)胞增生明顯,板層體豐富。 【結(jié)論】.對實驗性紫紺型肺血少動物模型的肺組織進行病理形態(tài)觀察和分析發(fā)現(xiàn),肺血流較正常減少和肺動脈低壓時肺小血管發(fā)育呈現(xiàn)明顯不良或退化,同時伴有外周肺小動脈數(shù)量減少,但是對肺泡的形態(tài)發(fā)育和數(shù)量影響不大。形態(tài)結(jié)構(gòu)異常影響器官的正常功能,說明保持正常的肺血流或盡早改善肺血少狀態(tài)是促進外周肺血管正常發(fā)育和改善全身狀況的關(guān)鍵。 第三部分.肺血少肺動脈低壓時肺組織形態(tài)變化的生物學(xué)基礎(chǔ) 【目的】.紫紺型肺血少先心病動物模型的肺血管和肺組織形態(tài)結(jié)構(gòu)與正常不同,即發(fā)生了細(xì)胞外基質(zhì)重塑。通過測量肺細(xì)胞外基質(zhì)中部分結(jié)構(gòu)性蛋白和細(xì)胞因子的變化,分析肺血減少時肺細(xì)胞外基質(zhì)重塑和形態(tài)結(jié)構(gòu)變化的生物學(xué)基礎(chǔ)。 【方法】.健康幼豬16只,分為紫紺肺血減少組(D組,n=8)和對照組(C組,n=8)。首先建立紫紺型肺血少先心病動物模型。兩組動物正常飼養(yǎng)2個月,麻醉和正中開胸,均取雙側(cè)肺組織,液氮冷凍保存。采用酶聯(lián)免疫反應(yīng)方法(Elisa),測量兩組肺組織中的基質(zhì)金屬蛋白酶(MMP-2、MMP-9)及基質(zhì)金屬蛋白酶抑制劑(TIMP-1),肺組織Ⅰ型和Ⅲ型膠原纖維(CoⅠ和CoⅢ)和肺血管內(nèi)皮細(xì)胞生長因子(VEGF)含量的變化。 【結(jié)果】.對照組(C組)8只均存活;肺血減少組(D組)存活6只,成功建立紫紺型肺血少先心病動物模型。酶聯(lián)免疫吸附反應(yīng)(Elisa)測量D組和C組肺組織中基質(zhì)金屬蛋白酶MMP-2的含量分別為56.6±20.3 ng/ml和77.6±20.7 ng/ml,二者有顯著性差異(P＜0.01),D組和C組肺組織中基質(zhì)金屬蛋白酶MMP-9的含量分別為4.7±1.0 ng/ml和5.8±1.0 ng/ml,二者有顯著性差異(P＜0.01);基質(zhì)金屬蛋白酶抑制劑(TIMP-1)在D組和C組的含量分別為14.5±3.4ng/ml和20.2±3.8ng/ml,二者有顯著性差異(P＜0.001)。D組和C組肺組織Ⅰ型(CoⅠ)膠原纖維含量分別為814.6±2006.8 ng/ml和3122.1±2865.1ng/ml,二者有顯著性差異(P＜0.01),D組和C組肺血管Ⅲ型(CoⅢ)膠原纖維含量分別為12.7±4.7 ng/ml和8.2±4.2 ng/ml,二者有顯著性差別(P＜0.01)。肺血管內(nèi)皮細(xì)胞生長因子(VEGF)在D組和C組的含量分別為13.6±3.0 ng/ml和18.8±3.5ng/ml,二者有顯著性差異(P＜0.001)。 【結(jié)論】.紫紺型肺血少型先心病動物肺細(xì)胞外基質(zhì)發(fā)生構(gòu)形重建,肺小血管發(fā)育不良或退化。同多流量型肺動脈高壓的機制相似,肺血減少時肺內(nèi)環(huán)境改變啟動各種調(diào)控因素,引起細(xì)胞外基質(zhì)中結(jié)構(gòu)性蛋白和細(xì)胞因子等成分變化是基質(zhì)重塑的基礎(chǔ)。
[Abstract]:The first part. Interventional and surgical techniques were combined to establish the pulmonary blood disease in young pigs with the animal model
[Objective]. Hybrid adopts a novel surgical intervention and the combination of the establishment of animal model of heart disease with pulmonary blood of young pioneers.
[method] using after birth from 1 to February in young pigs, weighing 8 ~ 12kg, divided into two groups, 8 rats in each group.A) normal control group (C-group, n=8): sham operation group, open chest surgery treatment; B group (D-group) with decreased pulmonary blood flow, n=8): right front third lateral intercostal thoracotomy through the right atrial surface into the dilator of artificial atrial septostomy; the left anterolateral third intercostal thoracotomy, the main pulmonary artery band set shrinkage. All animal normal feeding for 2 months. Then a median sternotomy, measuring cross pulmonary artery pressure; were smoking outside peripheral artery, right atrium and pulmonary vein, analysis, calculation of Qp/Qs of left atrial blood for blood gas drainage; peripheral venous blood measurement of hematocrit (HCT) and hemoglobin concentration (HB).
[results]. 8 rats in control group were alive, the average weight increased to 23.0kg. decreased pulmonary blood group survival was 6, the average weight increased to 20.6kg. decreased pulmonary blood flow and arterial oxygen partial pressure group (PaO_2), arterial oxygen saturation (SaO_2), HCT and HB were 52.9 mmHg, 85.6% 16.6g/dl., and 49.8% in control group PaO_2, SaO_2, HCT and HB were 118 mmHg, 98% 12.2g/dl., 37.9% and two groups of corresponding index showed significant difference (P < 0.001). By four points and blood gas values calculated, reduce pulmonary blood group Qp/Qs 0.54:1.
[Conclusion]. By using the Hybrid method of interventional and surgical combination, through artificial atrial septostomy and pulmonary artery banding, successfully established a novel pulmonary blood less cyanotic congenital heart disease animal model for the study of the pulmonary blood, cyanotic heart disease in clinical pathology and physiology provides a useful tool.
The second part. The pathological observation and analysis of lung tissue when pulmonary blood is reduced to pulmonary arterial pressure
[Objective]. We have established the animal model with cyanotic pulmonary blood, the semi quantitative analysis of morphological and pathological morphology of immature small pulmonary vessels were decreased pulmonary blood flow, developmental characteristics of immature pulmonary small vessels to reduce pulmonary blood.
[method]. 16 healthy piglets were divided into cyanosis group (group D, n=8) and control group (group C, n=8). First establish the animal model of pulmonary blood of the heart with the first part description methods. Two groups of normal animal feed for 2 months, anesthesia and thoracotomy. According to the animal biopsy steps were taken in the lower lobe, bilateral, organization of each one, 4% paraformaldehyde fixed after production, H-E and elastin +Van Gieson staining. The pathological morphological characteristics were observed in the two groups of animal pulmonary small vessels and lung parenchyma by using light microscopy and transmission electron microscopy; microscope image processing system, semi quantitative morphological analysis of pulmonary arterioles in the average film thickness (MT) and intima-media thickness (MT%), the average percentage of membrane area (MS) and membrane area percentage (MS%); the number of counts per unit area of pulmonary artery (APSC) and the number of alveoli per unit area (MAN) with a calculation; cut The proportion of the muscle type artery (CMA), the partial muscle artery (PMA) and the non muscle artery (NMA) in all the alveolar pulmonary artery (IAPA).
銆愮粨鏋溿，

本文編號：1357037