摘要
目的观察天麻提取的联苄类化合物32对鱼藤酮诱导的PC12细胞损伤的影响。方法4μmol/L鱼藤酮损伤PC12细胞48 h建立帕金森病体外模型。实验分为对照组、模型组、天麻联苄类化合物32不同浓度(0.1、1.0、10.0μmol/L)组,共5组。噻唑蓝[3-(4,5-dimethyhhyhhiazol-2-y1)-2,5-diphenyhetrazolium bromide,MTT]法检测PC12细胞存活率,Annexin V-FITC/PI染色流式细胞术检测PC12细胞凋亡,JC-1染色后,倒置显微镜下观察细胞线粒体膜电位水平,DCFH-DA荧光探针标记检测细胞内活性氧(reactive oxygen species,ROS)水平。结果与对照组相比,模型组能明显抑制细胞存活率(P<0.001),诱导细胞凋亡,同时降低线粒体膜电位,增加ROS含量(P<0.001)。与模型组相比,天麻联苄类化合物32不同浓度(0.1、1.0、10.0μmol/L)组的PC12细胞存活率显著增加(P<0.01,P<0.001),凋亡细胞比例减少,线粒体膜电位水平增高,细胞内ROS含量减少(P<0.001)。结论天麻联苄类化合物32对鱼藤酮损伤的PC12细胞具有保护作用,与其维持线粒体膜电位水平、减少氧化应激、抑制细胞凋亡有关。
Objective To observe the effects of a bibenzyl compound 32 from Gastrodia elata against rotenone damaged PC12 cells.Methods A cell model of Parkinson disease was established in PC12 cells by exposure to rotenone(4μmol/L)for 48 h.A total of five groups:control group,model group and the different concentrations(0.1,1.0,10.0μmol/L)of bibenzyl compound 32 from Gastrodia elata groups were set up.3-(4,5-dimethyhhyhhiazol-2-y1)-2,5-diphenyhetrazolium bromide(MTT)assay was used to measure the survival rate of the PC12 cells.Flow cytometry with Annexin V-FITC/PI staining was used to assess the apoptosis of PC12 cells.After JC-1 staining,mitochondrial membrane potential was observed under an inverted microscope.The intracellular reactive oxygen species(ROS)level was examined using DCFH-DA fluorescent probe.Results Compared with the control group,the model group could significantly inhibit cell survival rate(P<0.001),induce cell apoptosis,decrease mitochondrial membrane potential,and increase ROS content(P<0.001).Compared with model group,the survival rate of PC12 cells in the different concentrations(0.1,1.0,10.0μmol/L)of bibenzyl compound 32 from Gastrodia elata groups was significantly increased(P<0.01,P<0.001),the proportion of apoptotic cells decreased;mitochondrial membrane potential increased,intracellular ROS content decreased(P<0.001).Conclusion The bibenzyl compound 32 from Gastrodia elata has protective effect on PC12 cells damaged by rotenone,which may be related to the maintenance of the mitochondrial membrane potential,reducing oxidative stress and inhibiting apoptosis.
作者
吴苗苗
王惠芹
冯聚玲
苑玉和
石建功
陈乃宏
WU Miaomiao;WANG Huiqin;FENG Juling;YUAN Yuhe;SHI Jiangong;CHEN Naihong(State Key Laboratory of Bioactive Substances and Functions of Natural Medicine,Institute of Materia Medica,Chinese Academy of Medical Sciences,Beijing 100050,China;Hebei North University,Zhangjiakou,Hebei 075000,China;Hunan University of Chinese Medicine&Hunan Engineering Technology Center of Standardization and Function of Chinese Herbal Decoction Pieces,Changsha,Hunan 410208,China)
出处
《湖南中医药大学学报》
CAS
2022年第7期1076-1081,共6页
Journal of Hunan University of Chinese Medicine
基金
国家自然科学基金项目(U1402221,81773924,81803500)
湖南中医药大学中药一流学科(201803)。
关键词
天麻
帕金森病
鱼藤酮
细胞凋亡
氧化应激
Gastrodia elata
Parkinson diseases
rotenone
apoptpsis
oxidative stress
