乳腺癌前哨淋巴结新型荧光靶向示踪剂的动物模型研究

发布时间：2018-07-05 13:55

本文选题：显像剂 + 吲哚菁绿　；参考：《济南大学》2017年硕士论文

【摘要】：背景与目的:前哨淋巴结(Sentinel Lymph Node,SLN)活检已成为临床腋淋巴结阴性早期乳腺癌病人的标准处理模式,并对乳腺癌的分期和治疗方案的选择至关重要。该研究将吲哚菁绿(Indocyanine Green,ICG)和利妥昔单抗(Rituximab)进行偶联作为新型示踪剂,采用小鼠后肢SLN动物模型,模拟乳腺癌SLN活检术,探索其SLN的定位效应。方法:首先建立小鼠后肢淋巴引流模型。(1)Balb/c小鼠后肢脚背皮下注射不同剂量(0.48μg、0.24μg、0.12μg、0.06μg)的ICG-Rituximab,并应用荧光脉管系统成像仪连续观测乆窝淋巴结(SLN)至开始显像,后每隔5min观察一次持续至3h,记录SLN开始出现荧光显像的时间以及达到最佳显像效果所需的时间。3h脱颈椎处死小鼠(处死前5min于小鼠后肢脚背皮下相同位置注射亚甲蓝染料),解剖SLN及后续引流淋巴结,应用荧光脉管系统成像仪测量有无荧光显像,确定示踪剂的最佳注射剂量;(2)最佳注射剂量的ICG-Rituximab(0.12μg,ICG的含量)注射于小鼠后肢脚背皮下后分别于6h、12h、24h脱颈椎处死小鼠(处死前5min以相同方式注射亚甲蓝染料),解剖SLN及后续引流淋巴结,应用荧光脉管系统成像仪测量有无荧光显像。结果:定义乆窝淋巴结为小鼠后肢淋巴引流的SLN,髂淋巴结为次级淋巴结,主动脉旁或肾旁淋巴结为第三级淋巴结(个别小鼠主动脉旁淋巴结缺如,此时选取肾旁淋巴结为第三级淋巴结)。随着注射剂量的增加,SLN开始显像的时间与达到最佳显像效果所需的时间均逐渐缩短,次级及第三级淋巴结显像率逐渐升高。新型示踪剂的最佳注射剂量为0.12μg(ICG的含量),达到最佳显像效果的时间约为34min。观察至24h,SLN显像率维持在100%,次级及第三级淋巴结显像率由6h的0%和0%上升至24h的20%和10%。结论:本研究建立的小鼠后肢SLN动物模型设置简单,成本低,淋巴系统发达可以清晰揭示示踪剂在淋巴管和淋巴结中的引流情况。在小鼠SLN动物模型中ICGRituximab的最佳注射剂量为0.12μg(ICG的量),SLN活检时最佳的SLN显像时间窗为注射后34分钟至6小时,即能清晰定位SLN又无次级淋巴结显像,具有较高的临床应用价值。
[Abstract]:Background & objective: Sentinel Lymph NodeSLN biopsy has become the standard management model for early breast cancer patients with negative axillary lymph nodes, and is very important to the stage and treatment of breast cancer. In this study, indocyanine green (ICG) and Rituximab (Rituximab) were coupled as a new tracer. SLN animal model of mouse hind limb was used to simulate SLN biopsy of breast cancer to explore the localization effect of SLN. Methods: the lymphatic drainage model of the hind limb was established. (1) Balb / c mice were subcutaneously injected with different doses (0.48 渭 g / 0. 24 渭 g / g) of ICG-Rituximab. and the fluorescent vascular system was used to observe the lymph nodes (SLN) until the beginning of the imaging. After the 5min was observed every 3 hours, the time when the fluorescence imaging began to appear and the time required to achieve the best imaging effect were recorded. The mice were killed by removing the cervical vertebrae for 3 hours. (5min was injected in the same subcutaneous position on the back of the hind limbs of the mice before execution. Methylene blue dye), anatomic SLN and subsequent drainage lymph nodes, The fluorescence imaging system was used to measure the fluorescence imaging. The optimal dose of tracer was determined; (2) the optimal dose of ICG-Rituximab (0.12 渭 g ICG) was injected into the lower extremity of mice subcutaneously, and the mice were killed at 6h, 12h and 24h, respectively (5min was injected with methylene blue dye in the same way before execution). Subsequent drainage of lymph nodes, Fluorescence imaging system was used to measure fluorescence imaging. Results: the lymph nodes were defined as SLNs, iliac lymph nodes as secondary lymph nodes and para-aortic or para-renal lymph nodes as tertiary lymph nodes. The parrenal lymph nodes were selected as the third stage lymph nodes. With the increase of injection dose, the start time of SLN and the time needed to achieve the best imaging effect were shortened, and the secondary and tertiary lymph node imaging rates increased gradually. The optimal injection dose of the new tracer was 0.12 渭 g (ICG content), and the best imaging time was about 34 min. The SLN imaging rate at 24 h was maintained at 100 and the secondary and tertiary lymph node imaging rates increased from 0% and 0% at 6 h to 20% and 10% at 24 h. Conclusion: the SLN animal model of mouse hind limb established in this study has the advantages of simple setting and low cost. The developed lymphatic system can clearly reveal the drainage of tracer in lymphatic vessels and lymph nodes. The optimal injection dose of ICGRituximab in mouse SLN model was 0.12 渭 g (ICG). The best SLN imaging time window for SLN biopsy was 34 minutes to 6 hours after injection, which could locate SLN clearly without secondary lymph node imaging.
【学位授予单位】：济南大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.9;R-332

【参考文献】