1. 甘肃中医药大学 药学院,兰州 730000;2. 甘肃省中药药理与毒理学重点实验室,兰州 730000
1.Pharmacy School, Gansu University of Chinese Medicine, Lanzhou 730000, China. 2. Gansu Province Key Laboratory of Pharmacology and Toxicology of Traditional Chinese Medicine, Lanzhou 730000
目的 研究乳腺癌血行肺转移模型家兔血小板与凝血功能的变化规律。 方法 取健康雌性新西兰白兔 12 只,随机分为 2 组:正常组和模型组,模型组家兔左侧第 3 乳房乳垫注射 VX2 组织悬液,再经耳缘静脉注射 VX2 细胞悬液,构建乳腺癌血行肺转移家兔模型。 观察 42 d 内家兔精神状态、饮食情况,监测体重和肿瘤大小,检测血小板相关参数 PLT、MPV、PDW、PCT 和凝血相关指标 PT、APTT、TT、FIB 及 D-二聚体;实验终点 ELISA 法检测血清 CD63、CD62P、TXB2 及血浆 GMP-140 水平;取肿瘤组织和肺用 10%甲醛固定、石蜡包埋、切片、HE 染色,光学显微镜下观察肿瘤和肺组织形态变化。 结果 家兔乳垫注射 VX2 组织悬液附加静脉注射 VX2 细胞悬液,能成功构建乳腺癌血行肺转移家兔模型,肺转移率 100%,模型家兔血液处于高凝状态,14 ~ 42 d 内,PLT、MPV、PCT 均明 显增加;10 ~ 30 d 内,PT、APTT、TT 均明显缩短,且 FIB、D-二聚体含量均明显增加;血清 CD63、CD62P、TXB2 及血浆 GMP-140 水平明显增高;模型家兔乳腺中肿瘤细胞增多、排列疏松、炎性细胞浸润;肺组织可见肿瘤细胞团块伴出血、坏死。 结论 乳腺癌血行肺转移模型家兔血液环境异常,表现为血小板活化和血液高凝状态。
Objective To study the changes in platelets and coagulation function in a rabbit model of breast cancer lung metastasis. Methods Twelve female New Zealand white rabbits were randomly divided into two groups: normal group and model group. A VX2 breast cell suspension was injected into the third breast pad and injected intravenously through the ear marginal vein. The mental state, diet, body weight, and tumor size of the rabbits were observed over 42 days. Platelet- related parameters (PLT, MPV, PDW, and PCT) and coagulation-related indexes (PT, APTT, TT, FIB, and D-dimer) were detected at corresponding time points. The levels of CD63 and CD62P TXB2 in serum, and GMP-140 in plasma, were detected by ELISA. The tumor tissue and lung tissue were fixed with 10% formaldehyde, embedded in paraffin, sectioned, and stained with hematoxylin-eosin. Morphological changes in the tumor and lung tissue were observed under an optical microscope. Results The rabbit model of breast cancer lung metastasis was successfully established by injecting a VX2 tissue suspension into the breast pad and a VX2 cell suspension intravenously. The lung metastasis rate was 100%. The blood of the model rabbits was in a hypercoagulable state. The levels of PLT, MPV, and PCT increased significantly from 14 to 42 days, the PT, APTT, and TT indexes were significantly shortened, and the contents of FIB and D-dimer were significantly increased from 10 to 30 days. The levels of CD63, CD62P and TXB2 in serum, and GMP-140 in plasma, increased significantly. Tumor cells increased in number, appeared loosely arranged, and were infiltrated by inflammatory cells in the mammary gland of model rabbits. A tumor cell mass with hemorrhage and necrosis was observed in the lung tissue, which contained hemosiderin deposition. Conclusions The blood circulation in the rabbit model of breast cancer lung metastasis was abnormal, and this manifested as platelet activation and a hypercoagulable state in the blood.