腋窝淋巴结清扫术后上肢继发性淋巴水肿动物模型建立的实验研究

发布时间：2018-04-29 10:49

本文选题：继发性淋巴水肿 + 猕猴　；参考：《山东大学》2017年博士论文

【摘要】：目的:继发性上肢淋巴水肿(Secondary upper limb lymphedema,SULL)是乳腺癌术后常见并发症之一,是腋窝淋巴结清扫术和电离放射治疗后区域淋巴管损伤形成的慢性进行性淋巴系统病变,严重影响患者身心健康。虽然近些年乳腺癌患者中淋巴水肿的诊断和治疗已取得重要进展,但淋巴水肿仍是影响该疾病患者生活质量和身心健康的重要因素。目前许多研究者致力于乳腺癌患者术后SULL的研究,以期改善术后生活质量。SULL治疗研究的一个重要方面就是动物模型的建立。迄今,据报道已成功建立多种淋巴水肿的动物模型,包括啮齿动物和食肉动物。虽然在非灵长类动物模型中关于淋巴水肿的发病机制和治疗的研究取得了鼓舞人心的成果,但是非灵长类淋巴水肿动物模型建模成功率相对较低,而且更重要的是其与人类的肢体在生理、解剖和功能上存在着很大差异,并不能正确反映人类的生理和病理状态。基于上述原因,这些传统的动物模型并不能很好的复制人类上肢淋巴水肿的病理生理过程。本研究拟使用与人类生理、解剖和遗传极其接近的猕猴作为研究对象,探索建立SULL模型的可行性,以及建模评价指标。方法:健康雌性猕猴,年龄4-5岁,模拟实施标准的人类腋窝淋巴结清扫术,分别于术前2 周和术后4周联合使用放射技术对动物腋窝区域进行电离放射,作用于猕猴上肢,以破坏残存淋巴管。超声成像检查腋窝血管血流通畅情况,以排除血管因素所致水肿;分段测量前臂中部、肘不和上臂中部的周径变化;高分辨率核磁共振成像(magnetic resonance imaging,MRI)淋巴管造影观察上肢淋巴系统影像学改变;生物电阻抗分析(bioelectrical impedance analysis,BIA)监测上肢细胞外液体改变;常规病理观察淋巴管形态学改变;免疫组织化学染色检测淋巴上皮标志物表达以进一步确认淋巴管形态学变化。结果:1.上肢周径:干预侧肢体呈进行性水肿,并持续存在,24个月内,干预侧上肢/对侧上肢周径比率为100%-137%,手掌厚度增加约15%-40%,表现为典型的淋巴水肿特征;2.MRI淋巴造影:手术3个月后MRI显示上肢淋巴管出现明显变化。与对侧正常上肢相比,干预测上肢的淋巴干消失,淋巴管结构被雾状背景衬托下明亮的点状样式所取代。在所有动物中,淋巴水肿显示为筋膜外分布,并在T2加权像上呈高信号强度。正面3D扰相梯度回波高分辨率MRI和数字减影血管造影(digital subtraction angiography,DSA)淋巴管造影显像中,观察到网状分布的扩张淋巴管,其淋巴回流延迟,提示存在与梗阻相关的新血管生成;3.B1A显示:上肢细胞外液水分容量增加,与淋巴水肿的病理学改变呈线性相关;4.病理切片和免疫组化染色显示了动物模型淋巴管数量增多,管腔明显扩张,淋巴管的变化曲率呈淋巴水肿的典型病理学改变。结论:1.猕猴SULL模型的建立过程简单,容易成模。2.猕猴SULL动物模型以与人类更相似的遗传背景复制了该病的病理生理学特点。3.评价方法简便易行,该新型动物模型有助于增加我们对上肢获得性淋巴水肿的认识,并可促进新治疗方法的开发。
[Abstract]:Objective: secondary upper limb lymphedema (Secondary upper limb lymphedema, SULL) is one of the common complications after breast cancer surgery. It is a chronic progressive lymphatic disease caused by axillary lymph node dissection and ionizing radiation therapy, which seriously affects the physical and mental health of the patients. Although in recent years, the lymph nodes in the breast cancer patients are lymphatic. An important progress has been made in the diagnosis and treatment of edema, but lymphedema is still an important factor affecting the quality of life and physical and mental health of the patients. Many researchers are currently working on the study of postoperative SULL in patients with breast cancer, in order to improve the research of.SULL treatment for postoperative life quality is the establishment of animal models. It is reported that many animal models of lymphedema have been successfully established, including rodents and carnivorous animals. Although there are encouraging results in the study of the pathogenesis and treatment of lymphedema in the non primate model, the success rate of non primate lymphoedema animal model modeling is relatively low and more important. It is different from the human body in physiology, anatomy and function, and does not correctly reflect the physiological and pathological state of human beings. Based on the above reasons, these traditional animal models do not reproduce well the pathophysiological process of the human upper limb lymphedema. This research is intended to be used with human physiology, anatomy and heredity. The feasibility of establishing SULL model and modeling evaluation index were explored. Methods: healthy female rhesus monkeys, aged 4-5 years old, simulated the implementation of standard human axillary lymph node dissection, were used to ionize radiation in the axillary region of animals at 2 weeks before and 4 weeks after operation respectively. The upper limb was used to destroy the remaining lymphatic vessels. Ultrasound imaging examined the patency of the axillary vascular blood flow to exclude the edema caused by vascular factors; measure the middle of the forearm and the circumference of the middle of the upper arm in the middle of the forearm; the high-resolution magnetic resonance imaging (magnetic resonance imaging, MRI) lymphangiogram observed the imaging changes of the upper limb lymphatic system; Bioelectrical impedance analysis (BIA) was used to monitor the changes in the outer fluid of the upper extremity; the morphological changes of the lymphatics were observed by routine pathology; the expression of lymphoepithelial markers was detected by immunohistochemical staining to further confirm the morphological changes of the lymphatics. Results: 1. the circumferential diameter of the upper limb: the intervention side limbs were progressive edema and continued. Within 24 months, the ratio of the circumferential diameter of the upper limb / contralateral upper limb of the intervention side was 100%-137% and the palmar thickness increased by about 15%-40%, which showed typical lymphedema characteristics; 2.MRI lymphography: 3 months after the operation, the MRI showed obvious changes in the upper limb lymphatic vessels. Compared with the normal upper limbs, the lymphatic trunk of the upper limb disappeared and the lymphatic structure was foggy compared with the normal upper limb. In all animals, lymphedema is displayed outside the fascia and shows high signal intensity on the T2 weighted image in all animals. The reticular distribution of the positive 3D phase gradient echo high resolution MRI and the digital subtraction angiography (DSA) angiography (digital subtraction angiography, DSA) lymphangiography Dilated lymphatic vessels, with delayed lymphatic reflux, suggested the existence of new angiogenesis associated with obstruction; 3.B1A showed an increase in water capacity in the outer fluid of the upper limb and a linear correlation with the pathological changes of lymphedema; 4. pathological sections and immunohistochemical staining showed the increase in the number of lymphatics in the animal model, the obvious dilatation of the lumen and the change of lymphatic vessels. Typical pathological changes in the curvature of lymphedema. Conclusion: 1. the establishment of SULL model in rhesus monkeys is simple and easy to model.2. macaque SULL animal model to replicate the pathophysiological characteristics of the disease with the more similar genetic background of human beings. The.3. evaluation method is simple and easy. This new animal model helps to increase our upper limb acquired lymph nodes. The understanding of edema can promote the development of new treatment methods.

【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R737.9;R-332

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