α-倒捻子素通过激活ROS/p38 MAPK/Bax级联反应诱导B淋巴瘤Ramos细胞凋亡的机制研究  

Mechanism of α-mangostin-induced apoptosis in B lymphoma Ramos cells by activating ROS/p38 MAPK/Bax cascade reaction

作　　者：蔡紫微 孙毅松 汪林 廖玉娇 余欢 黄敏 李润滋 李敏惠 CAI Zi-wei;SUN Yi-song;WANG Lin;LIAO Yu-Jiao;YU Huan;HUANG Min;LI Run-zi;LI Min-hui(School of Basic Medicine,Chengdu Medical College;School of Bioscience and Technology,Chengdu Medical College;School of Pharmacy,Chengdu Medical College;Center of Scientific Research and Experiment,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院基础医学院 [2]成都医学院生物科学与技术学院 [3]成都医学院药学院 [4]成都医学院科研实验中心,成都610500

出　　处：《天然产物研究与开发》2022年第11期1832-1837,共6页Natural Product Research and Development

基　　金：四川省科技厅项目(2022NSFSC0690,2020YFS0321);国家级大学生创新创业项目(202213705004);四川省发育与再生重点实验室开放项目(SYS20-06)。

摘　　要：本文以B淋巴瘤Ramos细胞为研究对象,探讨α-倒捻子素(α-mangostin,α-Ma)对B淋巴瘤的增殖抑制作用并初步阐释其分子机制。首先通过CCK-8法探究α-Ma对Ramos细胞的增殖抑制作用;再利用倒置显微镜成像法观察α-Ma对Ramos细胞形态的影响;随后利用DCFH-DA、JC-1、Annexin V-FITC/PI荧光染色和流式细胞术检测α-Ma对Ramos细胞活性氧水平、线粒体膜电位、凋亡的影响;并通过蛋白免疫印迹技术测定α-Ma作用Ramos后细胞凋亡相关蛋白及信号通路蛋白的表达情况。CCK-8分析结果显示,α-Ma以药物浓度依赖和时间依赖的方式抑制Ramos细胞增殖,其24 h和48 h的IC值分别为14.84μmol/L和8.087μmol/L;倒置显微镜成像法发现α-Ma能减少Ramos细胞数量并诱导细胞形态发生凋亡样改变;流式细胞术检测发现,α-Ma能增加Ramos细胞内活性氧水平、降低细胞线粒体膜电位,同时诱导细胞凋亡;蛋白免疫印迹结果显示,α-Ma下调caspase-3/9的表达,上调cleaved caspase-3/9、cleaved PARP、Bax、Bim和p-p38 MAPK的表达。综上所述,α-Ma对B淋巴瘤Ramos细胞具有增殖抑制作用,其可能机制是α-Ma活化ROS/p38 MAPK/Bax级联反应诱导B淋巴瘤细胞凋亡。B lymphoma Ramos cells were used as the research object to explore the inhibitory effect of α-mangostin(α-Ma) on the proliferation of B lymphoma and explain its molecular mechanism.Firstly, the inhibitory effect of α-Ma on Ramos cells was investigated by the CCK-8 method.Then the effect of α-Ma on the morphology of Ramos cells was observed by inverted microscope imaging.DCFH-DA,JC-1,Annexin V-FITC/PI fluorescence dye, and flow cytometry were used to detect the effects of α-Ma on the level of reactive oxygen species, mitochondrial membrane potential, and apoptosis in Ramos cells.Western blot was used to detect the expression of apoptosis-related proteins and signal pathway proteins in Ramos cells treated with α-Ma.CCK-8 analysis showed that α-Ma inhibited the proliferation of Ramos cells in a concentration-dependent and time-dependent manner, and its ICvalues at 24 h and 48 h were 14.84 μmol/L and 8.087 μmol/L,respectively.Inverted microscope imaging revealed that α-Ma reduced the number of Ramos cells and induced apoptosis-like changes in cell morphology.Flow cytometry analysis showed that α-Ma increased the level of reactive oxygen species, decreased the mitochondrial membrane potential, and induced apoptosis in Ramos cells.Western blot showed that α-Ma could down-regulation the expression of caspase-3/9 and up-regulate the expression of cleaved caspase-3/9,cleaved PARP,Bax, Bim, and p-p38 MAPK.In conclusion, α-Ma can inhibit the proliferation of B lymphoma Ramos cells, and the possible mechanism is that α-Ma activated ROS/p38 MAPK/Bax cascade reaction to induce apoptosis of B lymphoma cells.

关 键 词：α-倒捻子素 B淋巴瘤 ROS p38 MAPK 线粒体途径 

分 类 号：R285[医药卫生—中药学]