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

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.