八宝丹体外抑制淋巴管生成的作用机制研究  

Study on the Mechanism of Babao Dan Inhibiting Lymphangiogenesis in Vitro

作　　者：关建华 逯遥 黄彬 兰炜兰 林久茂 GUAN Jianhua;LU Yao;HUANG Bin;LAN Weilan;LIN Jiumao(Academy of Integrative Medicine of Fujian University of Traditional Chinese Medicine,Fuzhou Fujian China 350122;Fujian Key Laboratory of Integrative Medicine on Geriatrics,Fuzhou Fujian China 350122)

机构地区：[1]福建中医药大学中西医结合研究院,福建福州350122 [2]福建省中西医结合老年性疾病重点实验室,福建福州350122

出　　处：《中医学报》2022年第1期135-141,共7页Acta Chinese Medicine

基　　金：福建省自然科学基金项目(2019J01355)。

摘　　要：目的:探讨八宝丹(babao dan, BBD)体外抑制淋巴管生成的作用机制。方法:将人淋巴内皮细胞(human lymphatic endothelial cells, HLECs)分为不同浓度的BBD组,即0 g·L^(-1)BBD组、0.25 g·L^(-1)BBD组、0.5 g·L^(-1)BBD组及0.75 g·L^(-1)BBD组。MTT法检测细胞增殖情况;Hoechs33258染色实验观察细胞凋亡情况;细胞划痕实验考察细胞损伤修复能力;Transwell实验考察细胞的迁移能力;管腔形成实验分析细胞淋巴管生成能力;Western Blot检测血管内皮生长因子-C(vascular endothelial growth factor-C,VEGF-C)、血管内皮生长因子受体-3(vascular endothelial growth factor receptor-3,VEGFR-3)、基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)和MMP-9蛋白表达水平。结果:BBD可显著降低HLECs增殖活力及迁移率(P<0.01);显著升高HLECs凋亡率(P<0.01);显著抑制HLECs损伤修复能力;显著降低HLECs管腔生成数量及淋巴管生成相关蛋白VEGF-C、VEGFR-3、MMP-2及MMP-9蛋白表达水平(P<0.05)。结论:BBD可促进HLECs凋亡,抑制其增殖、迁移及淋巴管生成,作用可能与下调VEGF-C、VEGFR-3、MMP-2和MMP-9蛋白表达水平有关。Objective: To investigate the inhibitory mechanism of Babao Dan(BBD) on lymphangiogenesis in vitro.Methods: Human lymphatic endothelial cells(HLECs) were divided into different concentrations of BBD groups, namely 0 g·L^(-1)BBD group, 0.25 g·L^(-1)BBD group, 0.5 g·L^(-1)BBD group and 0.75 g·L^(-1)BBD group.Cell proliferation was detected by MTT assay;Hoechs33258 staining was used to observe the apoptosis;cell scratch test was used to investigate the repair ability of cell damage;Transwell experiment was used to investigate the migration ability of cells;the ability of lymphangiogenesis was analyzed by lumen formation experiment;the protein expression levels of vascular endothelial growth factor-C(VEGF-C),vascular endothelial growth factor receptor-3(VEGFR-3),matrix metalloproteinase-2(MMP-2) and MMP-9 were detected by Western blot.Results: BBD could significantly reduce the proliferation activity and migration rate of HLECs(P<0.01),significantly increase the apoptosis rate of HLECs(P<0.01),significantly inhibit the damage repair ability of HLECs, and significantly reduce the number of lumen formation and the expression levels of lymphangiogenesis related proteins VEGF-C,VEGFR-3,MMP-2 and MMP-9(P<0.05).Conclusion: BBD can promote apoptosis, inhibit proliferation, migration and lymphangiogenesis of HLECs, which may be related to down regulating the expression levels of VEGF-C,VEGFR-3,MMP-2 and MMP-9 proteins.

关 键 词：八宝丹 淋巴管生成 VEGF-C VEGFR-3 MMP-2 MMP-9 

分 类 号：R285.5[医药卫生—中药学]