罗汉果苷Ⅴ对多巴胺能神经元炎症损伤的保护作用  

Protective effect of mogroside V on inflammatory damage of dopaminergic neurons

作　　者：蒋德旗 梁瑞兰 刘珂 梁焕葵 陈锦璐 陈泓睿 蒋丽林 JIANG De-qi;LIANG Rui-lan;LIU Ke;LIANG Huan-kui;CHEN Jin-lu;CHEN Hong-rui;JIANG Li-lin(Guangxi Key Laboratory of Agricultural Resources Chemistry and Biotechnology,College of Biology and Pharmacy of Yulin Normal University,Yulin 537000,China)

机构地区：[1]玉林师范学院生物与制药学院、广西农产资源化学与生物技术重点实验室,玉林537000

出　　处：《天然产物研究与开发》2024年第11期1931-1938,共8页Natural Product Research and Development

基　　金：玉林师范学院高层次人才科研项目(G2024ZK03)。

摘　　要：研究罗汉果苷Ⅴ(mogrosideⅤ,MogⅤ)对小胶质细胞激活所致多巴胺能神经元炎症损伤的保护作用及相关机制。多巴胺能神经元MN9D细胞与小胶质细胞BV-2在Transwell共培养,添加脂多糖激活小胶质细胞建立体外炎症损伤模型。将MN9D细胞分为对照组、模型组、MogⅤ组,其中MogⅤ组细胞在模型组基础上,添加50μmol/L MogⅤ作用48 h。流式细胞术检测细胞凋亡和细胞周期分布,JC-1法检测线粒体膜电位变化,酯化钙黄绿素与氯化钴共孵育法分析线粒体膜通透性转运孔的开放情况,利用荧光探针DCFH-DA分析细胞内活性氧(reactive oxygen species,ROS)水平,Western blot检测蛋白表达情况。结果显示,与对照组比较,炎症损伤使模型组多巴胺能神经元MN9D细胞的凋亡率明显升高,S期细胞比例明显降低,线粒体膜电位明显下降,线粒体膜通透性转运孔开放和ROS生成增多,SIRT3、超氧化物歧化酶2(superoxide dismutase 2,SOD2)及Bcl-2蛋白表达明显减少,Bax、Caspase-3蛋白表达显著增加,组间比较差异均具有统计学意义(P<0.05);与模型组比较,MogⅤ处理则使MogⅤ组多巴胺能神经元的凋亡率降低,S期细胞比例增加,线粒体膜电位上升,线粒体膜通透性转运孔开放和ROS生成减少,SIRT3、SOD2及Bcl-2蛋白表达显著增多,Bax、Caspase-3蛋白表达明显减少,组间比较差异均具有统计学意义(P<0.05)。以上结果表明,MogⅤ能够抑制小胶质细胞激活所致多巴胺能神经元的细胞凋亡和周期阻滞,对炎症损伤具有一定的缓解作用,机制可能与其促进SIRT3表达改善线粒体功能有关。This study aims to investigate the protective effects of mogroside V(Mog V)in dopaminergic neurons against the inflammatory damage caused by microglia activation and its related mechanism.Dopaminergic neurons(MN9D cells)and microglia(BV-2 cells)were co-cultured in Transwell plate,and lipopolysaccharide was added to activate microglia to establish an inflammatory model in vitro.MN9D cells were divided into the control group,model group and Mog V group.MN9D cells in the Mog V group were treated with 50μmol/L Mog V for 48 h based on the model group.Flow cytometry was used to test cell apoptosis rate and cell cycle distribution,JC-1 method was used to detect the changes in mitochondrial membrane potential,and the opening of mitochondrial permeability transition pores(mPTP)was analyzed by co-incubation of calcein-AM and CoCl 2.Fluorescent probe DCFH-DA was used to analyze the intracellular reactive oxygen species(ROS)levels and Western blot was used to test protein levels.The results showed that,compared with those of control group,inflammatory injury significantly increased the cell apoptosis rate,reduced the proportion of S-phase cells and mitochondrial membrane potential,increased the mPTP opening and ROS generation,decreased the protein expression of SIRT3,superoxide dismutase 2(SOD 2)and Bcl-2,and significantly increased the protein expression of Bax and Caspase-3 of dopaminergic neuron MN9D cells in the model group,and the differences between the two groups were statistically significant(all P<0.05).Compared with the model group,Mog V treatment decreased the apoptosis rate of dopaminergic neurons,increased the proportion of S-phase cells and mitochondrial membrane potential,reduced the mPTP opening and ROS production,significantly increased the protein levels of SIRT3,SOD 2 and Bcl-2,and significantly decreased the protein levels of Bax and Caspase-3 of MN9D cells in the Mog V group,and the differences between the two groups were statistically significant(all P<0.05).The above results indicate that Mog V can in

关 键 词：罗汉果苷Ⅴ 多巴胺能神经元 炎症 SIRT3 活性氧 

分 类 号：R966[医药卫生—药理学]