益气解毒方主要三萜类化合物抑制鼻咽癌CNE2细胞增殖效应的比较  

Comparison of Inhibitory Effects of Main Triterpenoids of Yiqi Jiedu Formula on Proliferation of Nasopharyngeal Carcinoma CNE2 Cells

作　　者：刘洁[1] 胡晶[1] 戴娜[1] 徐冰雁 罗晶婧 胡梅[1,2,3] 何迎春 LIU Jie;HU Jing;DAI Na;XU Bingyan;LUO Jingjing;HU Mei;HE Yingchun(Hunan University of Chinese Medicine,Changsha,Hunan 410208,China;Hunan Provincial Key Laboratory for the Prevention and Treatment of Ophthalmology and Otolaryngology Diseases with Traditional Chinese Medicine,Changsha,Hunan 410208,China;Hunan Provincial Ophthalmology and Otolaryngology Diseases Prevention and Treatment with Traditional Chinese Medicine and Visual Function Protection Engineering and Technological Research Center,Changsha,Hunan 410208,China)

机构地区：[1]湖南中医药大学,湖南长沙410208 [2]中医药防治眼耳鼻咽喉疾病湖南省重点实验室,湖南长沙410208 [3]湖南省中医药防治眼耳鼻咽喉疾病与视功能保护工程技术研究中心,湖南长沙410208

出　　处：《湖南中医药大学学报》2019年第11期1315-1320,共6页Journal of Hunan University of Chinese Medicine

基　　金：国家自然科学基金课题(81573721、81874408);湖南省自然科学基金项目(2016JJ6117、2019JJ4026);湖南省研究生创新项目(CX2018B482);湖南中医药大学研究生创新项目(2017CX33)

摘　　要：目的研究比较益气解毒方4种主要三萜类化合物齐墩果酸、熊果酸、茯苓酸、黄芪甲苷抑制鼻咽癌CNE2细胞增殖的效应。方法实时无标记细胞功能分析技术(real-time unlabeled cell function analysis technique,RTCA)动态监测不同浓度的4种益气解毒方三萜类化合物对体外培养的人鼻咽癌CNE2细胞生长的影响,高通量细胞成像多功能检测系统Cytation5(C5)监测不同化合物对鼻咽癌CNE2细胞形态的影响,Western Blot法检测不同化合物对鼻咽癌CNE2细胞增殖相关蛋白表达的影响。结果RTCA结果显示齐墩果酸、熊果酸、茯苓酸、黄芪甲苷均能够不同程度地抑制鼻咽癌细胞增殖,呈剂量依赖性,不同单体发挥最佳药效的时间不同。茯苓酸和黄芪甲苷起效较快,茯苓酸在处理后24 h达到最佳药效,IC50值为7.51μmol/L,黄芪甲苷持续作用36 h,抑制效果达到最佳,熊果酸在刺激28 h后IC50值最低,为0.70μmol/L,齐墩果酸后期作用效果较好,且所需浓度低。C5结果显示,与对照组相比,4种三萜类化合物在作用36 h后,CNE2细胞数量及形态均发生明显变化。Western Blot结果显示,与对照组相比,齐墩果酸、熊果酸、茯苓酸、黄芪甲苷能明显抑制CNE2细胞中增殖相关蛋白Survivin、XIAP、PCNA的表达。结论益气解毒方主要三萜类化合物均能抑制鼻咽癌细胞增殖,降低增殖相关蛋白Survivin、XIAP、PCNA的表达水平,但药物的作用时间和浓度均有差异。Objective To study and compare the inhibitory effects of the main triterpenoids of Yiqi Jiedu Formula including oleanolic acid,ursolic acid,pachymic acid and astragaloside iv on the proliferation of nasopharyngeal carcinoma CNE2 cells.Methods Real-time unlabeled cell function analysis technique(RTCA)was used to dynamically monitor the effects of 4 triterpenoids of Yiqi Jiedu Formula at different concentrations on the growth of human nasopharyngeal carcinoma CNE2 cells in vitro.Cytation 5(C5)was used to monitor the effects of different compounds on the morphology of nasopharyngeal carcinoma CNE2 cells.The effects of different compounds on the expression of proliferation-related protein of nasopharyngeal carcinoma CNE2 cells were detected by Western Blot.Results RTCA results showed that oleanolic acid,ursolic acid,pachymic acid and astragaloside iv could inhibit the proliferation of nasopharyngeal carcinoma cells in a dose-dependent manner,and the optimal time for different monomers was different.Pachymic acid and astragaloside iv arose faster,pachymic acid reached the best efficacy after 24 hours with treatment.IC50 was 7.51μmol/L,and the inhibitory effect of astragaloside iv was the best when it continued to function for 36 h.The IC50 of ursolic acid was the lowest after 28 hours of stimulation,which was 0.70μmol/L.Oleanolic acid had a better effect in the later stage and the action concentration was low.C5 results showed that compared with the control group,the number and morphology of CNE2 cells were significantly changed after 36 hours of treatment with 4 triterpenoids.Western Blot results showed that oleanolic acid,ursolic acid,pachymic acid and astragaloside could significantly inhibit the expression of proliferation-related proteins Survivin,XIAP and PCNA in CNE2 cells compared with the control group.Conclusion The main triterpenoids of Yiqi Jiedu Formula can inhibit the proliferation of nasopharyngeal carcinoma cells and reduce the expression of proliferation-related proteins Survivin,XIAP and PCNA,but t

关 键 词：益气解毒方 三萜类化合物 鼻咽癌 细胞增殖 

分 类 号：R285.5[医药卫生—中药学] R739.6[医药卫生—中医学]