李道睿 王苗苗 于明薇 林飞 张颖 李蒙 樊慧婷 郑琦 祁鑫 裴迎霞 张培彤 侯炜 林洪生

摘要：目的 觀察通络活血虫类药（全蝎、蜈蚣、守宫）对乏氧环境下肺癌血管生成相关细胞因子的影响。方法 采用低氧工作站体外模拟肿瘤乏氧微环境培养A549肺癌细胞，加入不同浓度全蝎、蜈蚣、守宫药物血清，MTT法检测细胞增殖，筛选最佳药物浓度及作用时间。A549肺癌细胞经3种药物血清处理后，收集细胞培养上清液，ELISA检测上清液中血管内皮生长因子（VEGF）、转化生长因子-β1（TGF-β1）、碱性成纤维细胞生长因子（bFGF）的含量。结果 3种药物血清对常氧及乏氧培养的A549肺癌细胞增殖均有一定的抑制作用。最终筛选7.5%浓度药物血清、作用24 h进行后续实验。3种药物血清较对照组A549细胞VEGF、TGF-β1、bFGF含量均降低（P<0.05，P<0.01）。结论 3种虫类药在常氧和乏氧状态下均有抑制癌细胞和对血管生成相关细胞因子调控的作用。

关键词：通络活血；虫类药；肺癌；乏氧

DOI：10.3969/j.issn.1005-5304.2017.09.010

中图分类号：R285.5 文献标识码：A 文章编号：1005-5304（2017）09-0039-04

Effects of Dredging Collaterals and Activating Blood Worm Chinese Materia Medica on Angiogenesis Related Factors of Lung Cancer in Hypoxic Environment LI Dao-rui1， WANG Miao-miao1，2， YU Ming-wei3， LIN Fei1， ZHANG Ying1， LI Meng1， FAN Hui-ting1， ZHENG Qi1， QI Xin1， PEI Ying-xia1， ZHANG Pei-tong1， HOU Wei1， LIN Hong-sheng1 （1. Guanganmen Hospital， China Academy of Chinses Medical Sciences， Beijing 100053， China； 2. Beijing University of Chinese Medicine， Beijing 100029， China； 3. Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital Medical University， Beijing 100010， China）

Abstract： Objective To observe the effects of dredging collaterals and activating blood worm Chinese materia medica on angiogenesis related factors of lung cancer in hypoxic environment. Methods The lung cancer A549 cells were cultured in vitro to simulate tumor hypoxia microenvironment by the hypoxia workstation， and different concentrations of Scorpio， Scolopendra and Gecko medicated serum were added. MTT method was used to detect cell proliferation and screen the best medicine concentration and duration of action. Lung cancer A549 cells were administrated by the three kinds of medicated serum， and cells were collected and supernatant was cultured. Contents of VEGF， TGF-β1， and bFGF were detected by ELISA. Results Three kinds of medicated serum had the inhibitory effect on both added normoxia and hypoxia in cultured A549 lung cancer cells. 7.5% concentration of medicated serum was selected， and 24 h later were used in later experiments. Scorpio， Scolopendra and Gecko medicated serum can more reduce the contents of VEGF， TGF-β1 and bFGF in the supernatant of A549 cell compared with the control group （P<0.05， P<0.01）. Conclusion Dredging collaterals and activating blood worm Chinese materia medica had inhibitory effect on cancer cells and the regulation of angiogenesis related cytokines in the condition of normoxia and hypoxia.

Key words： dredging collaterals and activating blood； worm Chinese materia medica； lung cancer； hypoxia

基金项目：国家自然科学基金（81302963）

通讯作者：林洪生，E-mail：dr.linhongsheng@163.com

肺癌是当今世界对人类健康与生命危害最大的恶性肿瘤之一，其发病率、死亡率居恶性肿瘤第1位。目前由于缺乏有效的治疗手段，不能很好地控制肺癌的转移，常常导致治疗的失败。因此，防止和阻断转移是肺癌治疗取得成功的关键。本课题组前期研究显示，肺癌转移的形成与肺癌患者普遍存在“络脉瘀滞”的现象有直接关系[1]，“络病”可能是肺癌发生转移的病机要点和关键环节，肿瘤乏氧微环境在这一过程中起着至关重要的作用。基于“虫蚁通络”理论，全蝎、蜈蚣、守宫等有通络活血作用的虫类药在临床中广泛应用于肺癌转移的防治中，其发挥作用的主要环节可能是改善肿瘤乏氧微环境。……