肝气郁滞型三阴性乳腺癌伴肝转移小鼠模型的构建与效果评价  

Construction and integration judgment for a mouse model of triple negative breast cancer with liver metastasis of liver qi depression type

作　　者：李洋 张志钦 韩超 王玮 王海永 李振祥 林家茂 薛洪范 LI Yang;ZHANG Zhiqin;HAN chao;WANG Wei;WANG Haiyong;LI Zhenxiang;LIN Jiamao;XUE Hongfan(Shandong Cancer Hospital and Institute,Shandong First Medical University and Shandong Academy of Medical Sciences,Jinan,Shandong250117,China;College of Medical Biology,Shandong First Medical University and Shandong Academy of Medical Sciences,Jinan,Shandong250118,China)

机构地区：[1]山东省肿瘤防治研究院(山东省肿瘤医院,放射免疫与分子影像实验室),山东第一医科大学(山东省医学科学院),山东济南250117 [2]山东省肿瘤防治研究院(山东省肿瘤医院,乳腺外科二病区),山东第一医科大学(山东省医学科学院),山东济南250117 [3]山东省肿瘤防治研究院(山东省肿瘤医院,胸部放疗一病区),山东第一医科大学(山东省医学科学院),山东济南250117 [4]山东省肿瘤防治研究院(山东省肿瘤医院,内三科),山东第一医科大学(山东省医学科学院),山东济南250117 [5]山东省肿瘤防治研究院(山东省肿瘤医院,中西医结合病区),山东第一医科大学(山东省医学科学院),山东济南250117 [6]山东省肿瘤防治研究院(山东省肿瘤医院,门诊管理部),山东第一医科大学(山东省医学科学院),山东济南250117 [7]山东第一医科大学(山东省医学科学院)生物医学科学学院,山东济南250118

出　　处：《中华肿瘤防治杂志》2024年第5期292-299,共8页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金青年项目(81904186);山东省中医药科技项目(Q-2022113)。

摘　　要：目的以乳腺癌原位移植模型为基础,通过慢性不可预知性应激联合孤养法构建肝气郁滞型三阴性乳腺癌伴肝转移小鼠模型。方法采用随机数字表法将6周龄BALB/c雌性小鼠40只分为4组,每组10只:对照组,正常饲养2周后予以乳腺垫接种4T1-luc乳腺癌细胞株,后期不给于刺激处理;模型组,慢性不可预知性应激(CUS)联合孤养;6-羟基多巴胺组(6-OHDA组),CUS联合孤养+6-OHDA;解郁消癥颗粒组,CUS联合孤养+解郁消癥颗粒。观察对比各组小鼠一般状况、旷场试验、蔗糖水偏嗜实验、促肾上腺皮质激素释放激素(CRH)、皮质酮(CORT)及去甲肾上腺素(NE)水平、肝脏HE染色和肝转移细胞凋亡水平,并行小鼠活体成像检测感兴趣区域(ROI)肿瘤荧光强度,磁共振成像检测肝脏转移灶数量。结果对照组、模型组、解郁消癥颗粒组和6-OHDA组糖水偏好、旷场试验中心区域的活动距离和穿梭次数得分,组间比较差异均有统计学意义(F=0.34,P<0.01;F=363.90,P<0.01;F=0.046,P<0.001)。模型组糖水偏好低于对照组,活动距离低于解郁消癥颗粒组,穿梭次数低于解郁消癥颗粒组,均P<0.01。对照组、模型组、解郁消癥颗粒组和6-OHDA组中血清CRH、CORT和NE表达量组间比较差异均有统计学意义(F=2501.00,P<0.01;F=155.30,P<0.001;F=149.80,P<0.01)。模型组CRH、CORT和NE表达量(87.71±5.33,198.90±5.64,371.00±16.08)均高于解郁消癥颗粒组(48.87±3.33,156.00±5.60,262.90±7.98),均P<0.01。解郁消癥颗粒组与模型组比较,ROI区域减少,肝转移数量减少,乳腺癌肝转移细胞凋亡指数增加,均P<0.01。结论以乳腺癌原位移植小鼠模型为基础,应用CUS联合孤养法能够良好模拟三阴性乳腺癌伴肝转移肝气郁滞证,可为探索肝气郁滞在三阴性乳腺癌伴肝转移中的证候实质及相关治疗药物的药理机制提供实验载体。Objective To establish a mouse model of liver qi depression triple negative breast cancer with liver metastasis by using chronic unpredictable stress combined with separation.Method In this experiment,6-week-old BALB/c female mice were selected.In the model group,4T1-luc breast cancer cell line was inoculated in the breast pad for 2weeks by chronic unpredictable stress(CUS)combined with solitary rearing method and continued to stimulate for 4weeks.In the same way,6-hydroxydopamine group(6-OHDA group)and Jie Yu Xiao Zheng Granule group were established.Observe and compare the general condition of mice in each group,open field test,sucrose water preference test,serum HPA axis related hormones(CRH,CORT,NE)level,liver HE staining,liver metastasis cell apoptosis level,and the tumor fluorescence intensity and the number of liver metastases detected by mouse living imaging.Result There was a statistically significant difference between the control group,model group,Jie Yu Xiao Zheng Granule group,and 6-OHDA group in sugar water preference,activity distance and shuttle frequency scores in the central area of the open field experiment(F=0.34,P<0.01;F=363.90,P<0.01;F=0.046,P<0.001).The preference for sugar water in the model group was significantly lower than the control group,the difference was statistically significant(P<0.01).The activity distance of the model group was significantly lower than the Jie Yu Xiao zheng Granule group,the difference was statistically significant(P<0.01).The shuttle frequency of the model group was significantly lower than the Jie Yu Xiao zheng Granule group,the difference was statistically significant(P<0.01).The average expression levels of serum CRH,CORT,and NE in the control group,model group,Jie Yu Xiao zheng Granule group,and 6-OHDA group were statistically significant(F=2501.00,P<0.01;F=155.30,P<0.001;F=149.80,P<0.01).The expression level of CRH in the model group was 87.71±5.33,which was significantly higher than the Jie Yu Xiao Zheng Granule group,which was 48.87±3.33,with a statis

关 键 词：三阴性乳腺癌 肝转移 肝气郁滞证 慢性不可预知性应激 动物模型 

分 类 号：R737.9[医药卫生—肿瘤]