地黄苷D对皮质酮诱导的PC-12细胞损伤的保护作用及机制研究  

Protective effect and mechanism of rehmannioside D on PC-12 cells injury induced by corticosterone

作　　者：张莉[1] 卢仁睿 王慧慧 李孟[1] 冯卫生[1,2] 郑晓珂 ZHANG Li;LU Ren-rui;WANG Hui-hui;LI Meng;FENG Wei-sheng;ZHENG Xiao-ke(School of Pharmacy,Henan University of Chinese Medicine,Zhengzhou 450046,China;Henan Engineering Technology Research Center for Chinese Medicine Development,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学药学院,河南郑州450046 [2]河南省中药开发工程技术研究中心,河南郑州450046

出　　处：《中草药》2022年第11期3385-3393,共9页Chinese Traditional and Herbal Drugs

基　　金：国家重点研发计划项目(2017YFC1702800);河南省重大科技专项(171100310500);河南省高层次人才特殊支持“中原千人计划”项目(ZYQR201810080);河南省教育厅河南省科技攻关项目(212102311106);河南省中医药科学研究专项课题(20-21ZY2151);国家留学基金委访问学者项目(201908410093);河南中医药大学2018年度博士科研基金资助项目(BSJJ2018-04)。

摘　　要：目的研究从地黄Rehmannia glutinosa中分离得到的地黄苷D对皮质酮诱导的肾上腺嗜铬细胞瘤细胞(PC-12细胞)的保护作用及机制。方法以500μmol/L皮质酮处理PC-12细胞24 h建立损伤模型,同时给予氟西汀和地黄苷D进行干预,采用MTT法检测细胞存活率;采用流式细胞术检测细胞凋亡、活性氧(reactive oxygen species,ROS)以及线粒体膜电位水平;采用In-Cell Western法检测细胞内凋亡相关蛋白半胱氨酸天冬氨酸蛋白酶-3(cystein-asparate protease-3,Caspase-3)、剪切型Caspase-3(cleaved Caspase-3)、B淋巴细胞瘤2(B-cell lymphoma 2,Bcl-2)和Bcl-2相关X蛋白(Bcl-2 associated X protein,Bax)表达;采用高内涵细胞成像系统检测细胞内脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)蛋白表达。结果地黄苷D明显提高皮质酮诱导的PC-12细胞存活率(P<0.05、0.01);抑制细胞凋亡和细胞内ROS水平(P<0.01);升高线粒体膜电位(P<0.01);下调细胞Bax/Bcl-2蛋白表达(P<0.05);上调BDNF蛋白表达(P<0.01)。酪氨酸激酶受体B(tyrosine kinase receptor B,Trk B)拮抗剂K252a能够拮抗地黄苷D对PC-12细胞凋亡的抑制作用。结论地黄苷D可能是通过激活BDNF-Trk B通路,抑制细胞凋亡通路,从而保护皮质酮诱导的PC-12细胞损伤。Objective To study the protective effect and mechanism of rehmangoside D isolated from Rehmannia glutinosa on PC-12 cells induced by corticosterone.Methods PC-12 cells were treated with 500μmol/L corticosterone for 24 h to establish an injury model,fluoxetine and rehmangoside D were given for intervention at the same time.Cell viability was detected by MTT method;Intracellular apoptosis,reactive oxygen species(ROS)and mitochondrial membrane potential levels were determined by flow cytometry;In-Cell Western method was used to detect intracellular apoptosis-related protein cysteine-aspartate protease-3(Caspase-3),cleaved Caspase-3,B-cell lymphoma 2(Bcl-2)and Bcl-2 associated X protein(Bax)expressions;High-content cell imaging system was used to detect intracellular brain-derived neurotrophic factor(BDNF)protein expression.Results Rehmangoside D significantly increased the survival rate of PC-12 cells induced by corticosterone(P<0.05,0.01),inhibited cell apoptosis and intracellular ROS level(P<0.01),increased mitochondrial membrane potential(P<0.01);Expression of Bax/Bcl-2 protein was down-regulated(P<0.05);BDNF protein expression was up-regulated(P<0.01).Tyrosine kinase receptor B(Trk B)antagonist K252a could antagonize the inhibitory effect of rehmangoside D on apoptosis of PC-12 cells.Conclusion Rehmangoside D may protect PC-12 cells from corticosterone-induced injury by activating BDNF-Trk B pathway and inhibiting apoptosis pathway.

关 键 词：地黄苷D 脑源性神经营养因子-酪氨酸激酶受体B通路 线粒体凋亡通路 皮质酮 抑郁症 

分 类 号：R285.5[医药卫生—中药学]