钩吻总生物碱及常绿钩吻碱的体外抗肿瘤作用及机制  

Antitumor effect and mechanism of total alkaloids of Gelsemium elegans and sempervirine in vitro

作　　者：陈惠娴 王文义 檀兴慧 李高攀 张晓琼 李德森[1] 吴水生[1] CHEN Huixian;WANG Wenyi;TAN Xinghui;LI Gaopan;ZHANG Xiaoqiong;LI Desen;WU Shuisheng(College of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Innovation and Transformation Center,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Dept.of Pharmacy,Quanzhou Guangqian Hospital,Fujian Quanzhou 362321,China)

机构地区：[1]福建中医药大学药学院,福州350122 [2]福建中医药大学科技创新与转化中心,福州350122 [3]泉州光前医院药剂科,福建泉州362321

出　　处：《中国药房》2023年第12期1437-1442,共6页China Pharmacy

基　　金：国家中医药管理局临床中药学高水平中医药重点学科建设项目(No.国中医药人教函〔2023〕85号);福建省科技计划项目(No.2020I0024,No.2020Y0051)。

摘　　要：目的探讨钩吻总生物碱(TA)和常绿钩吻碱(SPV)的体外抗肿瘤作用及机制。方法观察低、中、高浓度TA(50、100、200μg/mL)和SPV(10、30、50μmol/L)对人肝癌细胞(HepG2、Bel-7402)、人肺癌细胞(A549)、人结肠癌细胞(HCT-8)形态的影响并实时监测TA和SPV对4种肿瘤细胞的毒性;检测TA和SPV对HCT-8细胞上清液中胱天蛋白酶3(caspase-3)、caspase-9含量和细胞中磷酸化蛋白激酶B(p-Akt)(Thr308、Ser473)、B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)、生存素、C/EBP-同源蛋白(CHOP)、免疫球蛋白结合蛋白(Bip)、微管相关蛋白1轻链3Ⅱ(LC3Ⅱ)蛋白表达的影响。结果TA和SPV干预后的4种肿瘤细胞形态均出现体积缩小、细胞核皱缩的现象,细胞活性均出现不同程度的降低,其中TA和SPV对HCT-8细胞的抑制效果最好。TA和SPV干预后的HCT-8细胞上清液中caspase-3、caspase-9含量和细胞中Bax、CHOP、Bip、LC3Ⅱ蛋白表达水平均不同程度升高,p-Akt(Thr308、Ser473)、Bcl-2、生存素蛋白表达水平均不同程度降低。结论TA和SPV对上述4种肿瘤细胞均具有抑制作用,其对HCT-8细胞的抑制效果最好;二者对HCT-8细胞的作用机制可能与促进细胞凋亡、激活内质网应激和细胞自噬有关。OBJECTIVE To explore the antitumor effect and mechanism of total alkaloids of Gelsemium elegans(TA)and sempervirine(SPV)in vitro.METHODS The effects of low,medium and high concentrations of TA(50,100,200μg/mL)and SPV(10,30,50μmol/L)on the morphology of human hepatoma cells(HepG2,Bel-7402),human lung cancer cells(A549)and human colon cancer cells(HCT-8)were observed,and the toxicity of TA and SPV to four tumor cells was monitored.The effects of TA and SPV on the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells,the protein expressions of phosphorylated protein kinase B(p-Akt)(Thr308,Ser473),B-cell lymphoma 2(Bcl-2),Bcl-2-related X protein(Bax),survivin,C/EBP-homologous protein(CHOP),immunoglobulin binding protein(Bip)and microtubule-associated protein 1 light chain 3Ⅱ(LC3Ⅱ)in HCT-8 cells were detected.RESULTS After the intervention of TA and SPV,the volume reduction and nuclear shrinkage were founded in four tumor cells;the cell activity decreased to varying degrees,among which TA and SPV had the best inhibitory effect on HCT-8 cells.After the intervention of TA and SPV,the contents of caspase-3 and caspase-9 in the supernatant of HCT-8 cells,the protein expressions of Bax,CHOP,Bip and LC3Ⅱall increased to different degrees,while the protein expressions of p-Akt(Thr308,Ser473),Bcl-2 and survivin in HCT-8 cells all decreased to different degrees.CONCLUSIONS TA and SPV have inhibitory effects on the above four tumor cells,and the inhibitory effect on HCT-8 cells is the best.The mechanism of their action on HCT-8 cells may be related to promoting apoptosis,activating endoplasmic reticulum stress and autophagy.

关 键 词：钩吻总生物碱 常绿钩吻碱 抗肿瘤 细胞凋亡 内质网应激 细胞自噬 

分 类 号：R965[医药卫生—药理学]