阳虚证肺腺癌骨转移活体荧光成像模型的建立与评价

Establishment and evaluation of in vivo fluorescence imaging model in yang deficiency syndrome of lung adenocarcinoma with bone metastasis

目的:采用病证结合模型的方法建立阳虚证肺腺癌骨转移活体荧光成像模型,并评价其可行性。方法:将雌性BALB/c-nu小鼠随机分为假手术组、模型组、阳虚模型组,每组10只。模型组通过股骨骨髓腔种植表达绿色荧光蛋白人肺腺癌A549细胞建立肺癌骨转移活体荧光成像模型,阳虚模型组在模型组小鼠基础上采用臀部肌肉注射氢化可的松的方法建立裸鼠阳虚证模型;通过观察小鼠的一般情况与生存情况、小鼠体质量变化、小鼠肿瘤面积变化和阳虚症状评分,评价该模型的可行性。结果:①一般情况及生存情况:各组小鼠均未出现死亡及严重并发症。②小鼠体质量:实验期间,3组小鼠体质量比较差异均无统计学意义。③小鼠肿瘤面积:通过FluorVivo Model-100成像系统实时观察肿瘤生长情况,各组小鼠未出现其他部位和器官的转移,模型组和阳虚模型组的骨转移瘤面积随时间逐渐增大。实验结束时阳虚模型组肿瘤面积比模型组大,差异具有统计学意义(P<0.05)。④阳虚症状评价:实验结束时,阳虚模型组小鼠均出现了畏寒肢冷,活动减少、精神萎靡,饮食减少、皮毛苍白,大便清稀的阳虚症状,各症状评分明显高于模型组,差异具有统计学意义(P<0.01)。结论:采用"氢化可的松阳虚"裸鼠模型和"股骨骨髓腔种植转染绿色荧光蛋白肺腺癌A549细胞"骨转移活体荧光成像模型结合的造模方法安全有效,具有可行性。

Objective: To evaluation feasibility of in vivo fluorescence imaging model of yang deficiency syndrome of lung adenocarcinoma with bone metastasis that was established by combination of disease and syndrome method. Methods: Female BALB/c-nu mice were randomly divided into sham operation group, model group and yang deficiency syndrome model(YDSM)group, 10 in each group. Lung adenocarcinoma with bone metastasis mice models was established by intra-femur injection human lung adenocarcinoma A549 cells transfected with green fluorescent protein in model group. While YDSM was established by intrafemur injection combined with intramuscular injecting hydrocortisone. The survival situation, body weight, tumor area and yang deficiency syndrome were used to evaluate the model. Results: First, in general situation and survival situation, all mice were alive until the end without severe complications. The second, in body weight, the difference of mice body weight in each group was not significant during the test. The third, in tumor area, no metastasis of other parts and organs was observed in mice in each group by FluorVivo Model-100 imaging system. Tumor area of model group and YDSM group was increase over time. At the end of the experiment, mice tumor area of YDSM group was bigger than model group(P<0.05). The fouth, in symptom evaluation of yang deficiency syndrome: at the end of the experiment, the mice in YDSM group represented yang deficiency syndrome, such as being cold, reducing activity, being apathetic, less eating, pale fur and loose stools. The scores of yang deficiency syndrome in YDSM group were higher than those in model group(P<0.01). Conclusion: YDSM was established by intra-femur injection human lung adenocarcinoma A549 cells transfected combined with green fluorescent protein and intramuscular injecting hydrocortisone,showing high model quality and feasibility.