南蛇藤中萜类化合物雷酚萜促进胃癌细胞的凋亡研究  

Triptonoterpene,a terpenoid compound from Celastrus orbiculatus Thunb.promote apoptosis of gastric cancer cells

作　　者：欧诗雅 于彦威 罗园园 胡文杰 李沈忱 何正飞 戴小军[1,2] 王海波 张晓春[1,2] OU Shi-ya;YU Yan-wei;LUO Yuan-yuan;HU Wen-jie;LI Shen-chen;HE Zheng-fei;DAI Xiao-jun;WANG Hai-bo;ZHANG Xiao-chun(Medical College of Yangzhou University,The Key Laboratory of Syndrome Differentiation and Treatment of Gastric Cancer of the State Administration of Traditional Chinese Medicine,Yangzhou,Jiangsu 225001,China;College of Clinical Traditional Chinese Medicine of Yangzhou University,Yangzhou Hospital of TCM,Yangzhou,Jiangsu 225009,China)

机构地区：[1]扬州大学医学院,国家中医药管理局‘胃癌毒邪论治’重点研究室,江苏扬州225001 [2]扬州大学临床中医学院,扬州市中医院,江苏扬州225009

出　　处：《时珍国医国药》2023年第10期2383-2388,共6页Lishizhen Medicine and Materia Medica Research

基　　金：江苏省中医药科技发展计划重点项目(ZD202013);2018年度江苏省中医药领军人才项目;扬州大学大学生创新创业训练计划项目。

摘　　要：目的探究抗肿瘤中药南蛇藤中的二萜类化合物雷酚萜(Triptonoterpene)对胃癌细胞凋亡的影响。方法细胞计数试剂盒8(CCK-8)检测雷酚萜对胃癌细胞增殖的影响;观察雷酚萜对胃癌细胞形态变化的影响;凋亡实验评估雷酚萜促进胃癌细胞凋亡的情况;线粒体膜电位检测实验(JC-1)检测雷酚萜对胃癌细胞膜电位的影响;流式细胞仪检测雷酚萜对胃癌细胞内活性氧的影响;蛋白免疫印迹法(Western blot)用来检测B细胞淋巴瘤-2(Bcl-2)家族重要蛋白的表达以及细胞色素C(Cytochrome C)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)和切割型半胱氨酸天冬氨酸蛋白水解酶-3(cleaved Caspase-3)蛋白的表达。结果雷酚萜干预BGC-823细胞24 h和48 h后,细胞的增殖明显减慢;与对照组比较,雷酚萜高浓度组细胞内出现大量空泡,且呈蜂窝状;雷酚萜组胃癌细胞的凋亡率显著增加而且被PI染色的数量明显增多;同时雷酚萜组细胞中表示JC-1聚集的红色荧光减弱,表示JC-1单体的绿色荧光增强;另外雷酚萜组细胞内ROS的荧光强度显著增高;与对照组比较,雷酚萜干预胃癌细胞后,胞浆中Cytochrome C的表达增高,细胞中Bcl-2和B细胞淋巴瘤-xL(Bcl-xL)蛋白的表达降低,Bcl-2相关X蛋白(Bax)、Caspase-3和cleaved Caspase-3蛋白的表达升高。结论雷酚萜对胃癌细胞具有显著的促凋亡作用,很可能是南蛇藤中抗肿瘤有效成分的重要组成成分之一。Objective To explore the effect of Triptonoterpene,a diterpenoid compound from the anti-tumor Traditional Chinese medicine(TCM)Celastrus orbiculatus Thunb.,on the apoptosis of gastric cancer(GC)cells.Methods Cell Counting Kit-8(CCK-8)was used to detect the effect of Triptonoterpene on the proliferation of GC cells.The effect of Triptonoterpene on the morphological changes of GC cells was observed.Apoptosis experiments were used to evaluate the apoptosis of GC cells.Mito-chondrial membrane potential assay(JC-1)could determine the effect of Triptonoterpene on cell membrane potential of GC cells.The effect of Triptonoterpene on reactive oxygen species(ROS)in GC cells was detected by flow cytometry.Western blot-ting was used to detect the expression of Bcl-2 family proteins,Cytochrome C,Caspase-3 and cleaved Caspase-3.Results The proliferation of BGC-823 cells was significantly slowed down after Triptonoterpene intervention for 24 h and 48 h.Compared with the control group,there were a large number of vacuoles in the cells of the high concentration group of Triptonoter-pene,which were honeycomb-like.The apoptosis rate of GC cells in Triptonoterpene group was significantly increased and the number of PI staining was significantly increased.At the same time,the red fluorescence of JC-1 aggregation decreased and the green fluorescence of JC-1 monomer increased in Triptonoterpene group cells.In addition,the fluorescence intensity of ROS in Triptonoterpene group was significantly increased.Compared with the control group,the expression of Cytochrome c in the cytoplasm of GC cells increased after Triptonoterpene intervention,the expression of Bcl-2 and Bcl-xL protein in cells decreased,and the expression of Bax,Caspase-3 and cleaved Caspase-3 protein increased.Conclusion Triptonoterpene can induce apop-tosis of GC cells,which may be one of the effective components of Celastrus orbiculatus Thunb.as an anti-tumor Chinese medi-cine.

关 键 词：雷酚萜 萜类化合物 南蛇藤 胃癌 凋亡 

分 类 号：R285.5[医药卫生—中药学]