【作者】 汤海峰； 田冶； 王晓娟； 邱峰； 程光； 文爱东；

【Author】 TANG Hai-feng~1,TIAN Ye~(1,2),WANG Xiao-juan~3,QIU Feng~2,CHENG Guang~4,WEN Ai-dong~1 (1.Department of Pharmacy,Xijing Hospital,Fourth Military Medical University,Xi’an 710032;2. Department of Phytochemistry,Shenyang Pharmaceutical University,Shenyang 110015;3.Department of Pharmacy,School of Stomatology,Fourth Military Medical University,Xi’an 710032;4.Department of Neurosurgery,Xijing Hospital,Fourth Military Medical University,Xi’an 710032)

【机构】 第四军医大学西京医院药剂科； 沈阳药科大学中药学院天然药物教研室； 第四军医大学口腔医学院药剂科； 第四军医大学西京医院脑科医院；

【摘要】 目的研究中药九节龙中具有抗恶性胶质瘤活性的三萜皂苷成分。方法应用多种色谱手段和波谱方法分离和鉴定化合物。采用MTT法测定肿瘤细胞毒性,流式细胞术、透射电镜、Hoechst 33342荧光检测、DNA Ladder、Western blot等技术研究凋亡及其信号途径。结果分离鉴定了10种三萜皂苷类化合物,包括6个新化合物,分别命名为ardipusillosidesⅢ(1),Ⅳ(2)和ardipusillosides A～D(3～6),4个已知化合物为ardisiacrispin B(7),ardisiacrispin A(8),ardipusillosideⅠ(9)和ardipusillosideⅡ(10),其中7和8首次从本种得到。除皂苷6无活性、皂苷8具有中等强度活性外,其余8种皂苷均对人恶性胶质瘤显示显著细胞毒性,并均不影响人原代星形胶质细胞的生长。1可诱导人恶性胶质瘤U251MG细胞凋亡,机制研究表明涉及到BAD介导的细胞内凋亡信号途径以及caspase-8和caspase-3介导的细胞外凋亡信号途径。结论皂苷3的分子中苷元通过1个丙三醇基与糖醛酸连接,是三萜皂苷中非常罕见的结构特征;皂苷5和6的苷元以及皂苷4的30位羧基连接阿拉伯糖的结构特征,均为首次在本属(紫金牛属)植物中发现;皂苷1的苷元和皂苷2的六糖链结构在本属中也比较少见。这些皂苷显著的肿瘤细胞毒性及对其具体作用机制的研究结果为进一步开发抗恶性胶质瘤新药提供了有价值的先导化合物和科学依据。更多还原

【Abstract】 Objective To study the anti-glioblastoma triterpenoid saponins from Ardisia pusilla.Methods A combination of multi-chromatography as well as spectral analysis were employed for the separation and determination of the compounds from the plant.The cytotoxicities against human glioblastoma U251MG cells were determined using the MTT assays.The ability of ardipusillosideⅢto induce apoptosis in U251MG cells, as well as the involvement of apoptotic signaling pathways were also studied.Results Ten triterpenoid saponins(1-10) were obtained and elucidated,among which six are new compounds and named ardipusillosidesⅢ(1),Ⅳ(2) and ardipusillosides A-D(3-6).Nine saponins showed in vitro cytotoxicity against U251MG cells.Both the intrinsic pathway of apoptosis,mediated by BAD dephosphorylation and cleavage,and the extrinisic pathway of apoptosis,mediated by caspase-8 and caspase-3 activation,were involved in ardipusillosideⅢ-induced apoptosis.Conclusion The structural features of new saponins 1-6 are relative rare among triterpenoid glycosides or saponins from genus Ardisia.Our studies focused on cytotoxic saponins of A.pusilla have provided valuable leads for pursuing new anti-glioblastoma drugs,and established a homebase for further development of the title plant with abundant resources in Southern China.更多还原