乳移平调控前转移微环境抑制乳腺癌肺转移的实验研究  

Ruyiping Inhibited Pulmonary Metastasis of Breast Cancer by Regulating the Formation of Pre-metastasis Microenvironment

作　　者：叶依依[1] 刘胜[1,2] 孙霃平[2] 吴春宇[2] 

机构地区：[1]上海中医药大学附属龙华医院中医外科研究所,上海200032 [2]上海中医药大学附属龙华医院外七科,上海200032

出　　处：《中国中西医结合杂志》2017年第1期86-93,共8页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81102597)

摘　　要：目的观察抗乳腺癌复发转移方剂乳移平对乳腺癌前转移肺微环境的影响,探讨其可能机制。方法动物实验分两部分,第一部分是乳腺癌前转移时间的确定,第二部分是乳移平对前转移微环境的影响。第一部分实验24只BALB/c小鼠,取对数生长期4T1细胞配成细胞悬液,血细胞计数板计数,细胞浓度调至1×106细胞/m L,无菌条件下接种于24只BALB/c小鼠胸壁右侧第四乳头的脂肪垫上,每只0.1 m L。第二部分动物实验60只BALB/c小鼠,采用随机数字表法分成5组,分别为空白组、模型组、乳移平低、中、高剂量组,每组12只。造模方法同上,各组于接种次日开始给药。乳移平低、中、高剂量组给予5.13、10.26及20.52 g/(kg·d)生药量灌饲,每天给药1次,连续14天。空白组和模型组等体积生理盐水灌饲。动物实验第一部分于10、14、18、22天,每次脱颈处死6只,观察肺转移情况。动物实验第二部分观察各组肺组织的镜下形态(光镜)和肺血管超微结构(电镜),测定各组血管的通透性(依文思蓝法),研究乳移平对前转移微环境形成的影响。通过Western blot和Real time PCR法测定乳移平对前转移因子血管生成素(angiogenin,Angpt)2、血管内皮生长因子(vascular endothelial growth factor,VEGF)、白介素(Interleukin,IL)-6和IL-1β表达的影响。结果肺转移还未发生的乳腺癌前转移时期的建模时间定为14天。与模型组比较,乳移平中、高剂量组对瘤重、瘤体积都有显著的抑制作用(P<0.05,P<0.01),乳移平对肿瘤的重量和体积的抑制作用随着乳移平剂量的增加而增加,呈现明显的剂量依赖关系(P<0.05,P<0.01)。模型组与乳移平低剂量组都有淋巴细胞浸润的发生,而乳移平中、高剂量组光镜结果发现肺组织形态与空白组无区别。肺部的血管网是由连续的、密集的毛细血管组成。空白组肺脏毛细血管结构正常。模型组血管壁不像空白组平整、规则,�Objective To observe the effect of Ruyiping （RYP, a recipe for fighting against re- currence and metastasis of breast cancer） on pre-metastatic microenvironment, and to study its possi- ble mechanism. Methods The experiment was divided into two parts. The 1st part lies in setting the pre-cancerous transfer, and the 2nd part lies in the effect of RYP on pre-metastatic microenvironment. There were 24 BALB/c mice in the 1st part. Logarithmic phase 4T1 cells were dispensed into cell suspension. Blood cells were counted by blood cell counter. Then they were injected into the 4th mammary fat pad of the 24 BALB/c mice under aseptic condition （1 xl06cells/mL, 0.1 mL for each mouse）. There were 60 BALB/c mice in the 2nd part. They were divided into the blank group, the model group, low, middle, high dose RYP groups by random digit table, 12 in each group. The modeling method was the same as men- tioned above. Medication was started from the 2nd day of inoculation. Mice in low, middle, high dose RYP groups were administered with 5.13, 10.26,20.52 g/kg RYP crude drugs per day by gastrogavage, once per day for 14 successive days. Equal volume of normal saline was administered by gastrogavage to mice in the blank group and the model group. Six mice were sacrificed at day 10, 14, 18, and 22, respec- tively in the 1st part of the experiment. The pulmonary metastasis was observed. The histology and mi- cromorphology of lung tissues were observed under light microscope and electron microscope/transmis- sion electron microscopy （TEM） in the 2nd part of the experiment. The relative pulmonary vascular per- meability was determined by Evans blue. The effect of RYP on the formation of pre-metastatic microenvi- ronment was observed. The levels of angiogenin2 （Angpt2）, vascular endothelial growth factor （VEGF）, IL6 and ILl 13 were detected by Western blot and Real time PCR. Results The period from day 0 to day 14 was considered to be the pre-metastatic phase. Compared with the model group, significant inhibition on t

关 键 词：乳移平 乳腺癌 肺转移 前转移微环境 

分 类 号：R285.5[医药卫生—中药学]