摘要
目的研究碱制法炮制前后斑蝥对人肺癌A549细胞增殖、迁移、侵袭、凋亡的影响。方法提取生品斑蝥中的斑蝥素和碱制斑蝥中的斑蝥酸钠。以人非小细胞肺癌A549细胞为研究对象,生斑蝥与碱制斑蝥处理细胞后,采用CCK-8法检测药物对细胞增殖活性的影响,划痕实验观察细胞迁移作用,Transwell实验观察细胞侵袭作用,WB观察药物对A549细胞MMP1、MMP2蛋白表达的影响,ELISA法检测细胞炎症因子IFN-γ、IL-1β和TNF-α水平,AnnexinV/PI染色观察细胞凋亡情况。结果生斑蝥与碱制斑蝥均可使A549细胞活力明显降低;生斑蝥与碱制斑蝥均能抑制A549细胞迁移,且生斑蝥高浓度组划痕愈合程度最低,与生斑蝥高浓度组相比,差异具有显著性(P<0.01);碱制斑蝥较生斑蝥抑制A549细胞侵袭作用更明显,差异具有显著性(P<0.01);且生斑蝥与碱制斑蝥均可抑制MMP1和MMP2蛋白的表达,显著上调炎症因子IFN-γ水平,但对IL-1β、TNF-α含量则没有显著影响,Annexin V/PI双染荧光拍照发现生斑蝥组与碱制斑蝥组红绿荧光强度均增加,且碱制斑蝥组荧光增加更明显。结论碱制法炮制后斑蝥的抗癌作用显著提高,具有抑制A549细胞增殖,侵袭和迁移的作用,其机制可能与下调MMP1和MMP2的表达,促进凋亡及上调炎症因子IFN-γ水平有关。
Objective To assess the changes in the effects of cantharides after alkaline processing on proliferation,migration,invasion,and apoptosis of human lung cancer A549 cells.Methods Human non-small cell lung cancer A549 cells were treated with cantharis extract(CTE)from raw cantharides and alkali processed cantharis extract(ACE).The proliferation of the cells was detected with CCK-8 assay,and the cell migration and invasion were assessed using wound healing assay and Transwell assay,respectively.The expressions of MMP1 and MMP2 in the cells were detected usingWestern blotting,the contents of IFN-γ,IL-1βand TNF-αwere measured with ELISA,and cell apoptosis was analyzed with annexinV/PI fluorescent staining.Results Both CTE and ACE significantly reduced the viability and inhibited the migration of A549 cells,and high-dose ACE produced a significantly stronger inhibitory effect on cell migration than high-dose CTE(P<0.01).ACE showed more potent inhibitory effect than CTE on the invasion of A549 cells(P<0.01).Both CTE and ACE inhibited the expressions of MMP1 and MMP2 and up-regulated the level of IFN-γwithout significantly affecting the levels of IL-1βand TNF-α.Annexin V/PI staining showed that both CTE and ACE caused apoptosis of A549 cells,but ACE had a stronger proapoptotic effect.Conclusion Processing with sodium hydroxide can significantly improve the antitumor activity of cantharides,which inhibits the proliferation,migration and invasion of A549 cells possibly by down-regulating the expressions of MMP1 and MMP2,promoting apoptosis and increasing the level of IFN-γ.
作者
李娴
李姗姗
庞金龙
黄富豪
郭滨
刘浩
LI Xian;LI Shanshan;PANG Jinlong;HUANG Fuhao;GUO Bin;LIU Hao(School of Pharmacy,Bengbu Medical College,Bengbu 2330302,China;Anhui Provincial Key Laboratory of New Technology of Chinese Herbal Pieces Manufacturing,Bozhou 236800,China;Postdoctoral Workstation of Anhui Xiehecheng Pharmaceutical Yinpian Co Ltd,Bozhou 236800,China)
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2020年第9期1332-1339,共8页
Journal of Southern Medical University
基金
国家自然科学基金(81372899)
安徽省自然科学基金(1908085QH37)
安徽省教育厅高校优秀青年人才支持计划项目(gxyq2019037)
蚌埠医学院横向课题(2019-BYHX-03)。
关键词
碱制斑蝥
凋亡
炎症因子
迁移
侵袭
alkaline processed cantharis
apoptosis
inflammatory factors
migration
invasion
