无花果果实及乳浆对人黑色素瘤A375细胞的抑制作用及其作用机制研究  

Inhibitory Activity and Mechanism of Fig Fruit and Latex on Human Melanoma A375 Cells

作　　者：李保宏 李忠原 刘苗苗 田景振[1] 张晓 崔清华[2,3] LI Baohongi;LI Zhongyuan;LIU Miaomiao;TIAN Jingzhen;ZHANG Xiao;CUI Qinghua(College of Pharmacy,Shandong University of Traditional Chinese Medicine,Jinan 250355 Shandong,China;Innovation Research Institute of Traditional Chinese Medicine,Shandong University of Traditional Chinese Medicine,Jinan 250355 Shandong,China;Qingdao Academy of Chinese Medicinal Sciences,Shandong University of Traditional Chinese Medicine,Qingdao 266112 Shandong,China)

机构地区：[1]山东中医药大学药学院,山东济南250355 [2]山东中医药大学中医药创新研究院,山东济南250355 [3]山东中医药大学青岛中医药科学院,山东青岛266112

出　　处：《中药新药与临床药理》2022年第1期39-49,共11页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：山东省自然科学基金面上项目(ZR2020MH383);山东省重点研发计划(重大科技创新工程)项目(2020CXGC010505,2021CXGC010511)。

摘　　要：目的考察无花果实及乳浆不同极性部位的体外抗黑色素瘤活性,并探讨其作用机制。方法采用溶剂提取法制得无花果提取物,系统溶剂萃取法得乳浆不同极性部位和醇沉纯化部位供试品;采用MTT法、划痕实验、Transwell侵袭实验和流式细胞仪凋亡检测考察供试品对人恶性黑色素瘤细胞A375的增殖、迁移、侵袭和凋亡影响;构建A375细胞和人脐静脉内皮细胞HUVEC的共培养体系,通过侵袭实验、成管实验体外模拟肿瘤细胞对内皮血管形成;ELISA法测定相关细胞因子水平。结果无花果提取物和乳浆乙酸乙酯部位抑制肿瘤细胞A375的迁移和侵袭(P<0.05,P<0.01,P<0.001),降低A375细胞的超氧化物歧化酶(SOD)及乳酸脱氢酶(LDH)含量(P<0.001),提高一氧化氮(NO)含量(P<0.05,P<0.01,P<0.001);乳浆醇沉部位抑制A375细胞迁移和侵袭(P<0.01,P<0.001),对A375细胞的直接抑杀和促凋亡效果较好;共培养后HUVEC细胞侵袭能力提升,血管内皮生长因子(VEGF)、血管内皮素1(ET-1)、内皮型一氧化氮合酶(eNOs)分泌增多(P<0.001),且供试品(200~400μg·mL;)给药后可明显逆转(P<0.05,P<0.001)。结论无花果提取物及乳浆乙酸乙酯、醇沉部位具有抑制A375细胞增殖、迁移和侵袭作用,从能量代谢方面抑制黑色素瘤生长,并可能通过eNOs/VEGF/ET-1途径抑制肿瘤的新生血管形成。Objective To investigate the anti-melanoma activity of fig extract and different polar parts of its latex in vitro and explore the mechanism. Methods Fig extract was prepared by solvent extraction,different polar parts of latex and purified parts of alcohol-sedimentation fraction were obtained by systematic solvent extraction. The effects of extracts on the proliferation, migration, invasion, and apoptosis of A375 cells were investigated by MTT method,scratch test,Transwell invasion test and flow cytometer apoptosis experiment,respectively. The co-culture system of A375 cells and HUVEC cells was constructed. Endothelial angiogenesis of A375 cells was simulated by invasion test and tube formation assay in vitro, and the levels of related cytokines were measured by ELISA to investigate the mechanism of action. Results Fig extract and ethyl acetate fraction of latex inhibited the migration and invasion of A375 cells in a concentration-dependent manner(P<0.05,P<0.01,P<0.001),decreased the contents of SOD and LDH(P<0.001),and increased the content of NO in A375 cells(P<0.05,P<0.01,P<0.001). Alcohol-sedimentation fraction of latex inhibited the migration and invasion of A375 cells(P<0.01,P<0.001),had better effect on the direct inhibition and promotion of apoptosis of A375 cells. After co-culture,the invasive ability of HUVEC cells increased,the secretion of VEGF,ET-1 and eNOs increased(P<0.001),but it can be significantly reversed after administration of extracts(200~400 μg·mL;)(P<0.05,P<0.001). Conclusion Fig extract, ethyl acetate fraction and alcohol-sedimentation fraction of latex can inhibit the proliferation,migration,and invasion of A375 cells,inhibit the growth of melanoma in terms of energy metabolism. Fig extract and ethyl acetate fraction of latex may inhibit tumor neovascularization through eNOs/VEGF/ET-1 pathway.

关 键 词：无花果 无花果乳浆 黑色素瘤 A375细胞 血管生成 eNOs/VEGF/ET-1 

分 类 号：R285.5[医药卫生—中药学]