丁基苯酞通过PRDM5改善氧化应激引起的神经元细胞凋亡  

3-butylphthalideimproves neuronal apoptosis induced by oxidative stress through PRDM5

作　　者：何康 高永涛 徐晨阳 段国庆 HE Kang;GAO Yong-tao;XU Chen-yang;DUAN Guo-qing(Department of Neurosurgery,Huaihe Hospital of Henan University,Kaifeng 475000,Henan Province,China)

机构地区：[1]河南大学淮河医院神经外科,河南开封475000

出　　处：《中国临床药理学杂志》2024年第1期17-21,共5页The Chinese Journal of Clinical Pharmacology

基　　金：河南省科技厅攻关计划基金资助项目(232102310356)。

摘　　要：目的探讨丁基苯酞对氧化应激诱导的神经元细胞PC-12凋亡的影响及其作用机制。方法将PC-12细胞分成Vector组[二甲基亚砜(DMSO)+转染vector质粒]、丁基苯酞+vector组(DMSO+25μmol·L^(-1)丁基苯酞+转染vector质粒)、PRDM5组[DMSO+转染PR结构域蛋白5(PRDM5)过表达质粒]、丁基苯酞+PRDM5组(DMSO+25μmol·L^(-1)丁基苯酞+转染PRDM5过表达质粒)。用流式细胞术检测神经元细胞的凋亡水平;用蛋白质印迹法检测切割的胱天蛋白酶3(Cleaved-caspase 3)、B细胞淋巴瘤因子2(Bcl-2)、Bcl-2相关X(Bax)的表达水平;用分子对接分析丁基苯酞与PRDM5的结合位点。结果Vector组、丁基苯酞+vector组、PRDM5组、丁基苯酞+PRDM5组的细胞凋亡水平分别为(63.52±5.72)%、(20.48±3.56)%、(79.48±8.13)%和(22.58±3.01)%;Cleaved-caspase 3表达水平分别为0.89±0.09、0.29±0.03、1.12±0.09和0.31±0.05;Bcl-2表达水平分别为0.31±0.02、0.79±0.05、0.12±0.01和0.89±0.11;Bax表达水平分别为0.83±0.08、0.25±0.03、1.03±0.11和0.27±0.03。Vector组的上述指标与PRDM5组比较,差异均有统计学意义(均P<0.05)。丁基苯酞与PRDM5存在结合(自由能为-12.55 kcal·mol-1),结合位点为K413和D414。结论丁基苯酞与PRDM5的K413R和D414A结合,抑制了PRDM5的功能进而改善了氧化应激引起的神经元细胞凋亡。Objective To investigate the effects of 3-Butylphthalideon oxidative stress-induced apoptosis of neuronal cells PC-12 and its mechanism.Methods PC-12 cells were divided into four groups,Vector group:DMSO treated and transfected with vector plasmid;3-butylphthalide+vector group:DMSO+25μmol·L^(-1)3-butylphthalidewas treated with vector plasmid was transfected;PRDM5 group:PR domain zinc finger protein 5(PRDM5)overexpression plasmid was transfected with DMSO;3-butylphthalide+PRDM5 group:Treated with DMSO+25μmol·L^(-1)3-butylphthalide and transfected with PRDM5 overexpression plasmid.Apoptosis levels of neurons were detected by flow cytometry;Cleaved cysteine aspartate proteinase 3(Cleaved-caspase 3),B cell lymphoma-2(Bcl-2)and Bcl-2 associated X(Bax)expression levels were detected by Western blotting;binding sites of 3-Butylphthalideand PRDM5 were analyzed by molecular doc ki ng.Results The apoptosis levels of vector group,3-butylphthalide+vector group,PRDM5 group and 3-butylphthalide+PRDM5 group were(63.52±5.72)%,(20.48±3.56)%,(79.48±8.13)%and(22.58±3.01)%;Cleaved-caspase 3 expression levels were 0.89±0.09,0.29±0.03,1.12±0.09 and 0.31±0.05;the expression levels of Bcl-2 were 0.31±0.02,0.79±0.05,0.12±0.01 and 0.89±0.11;the expression levels of Bax were 0.83±0.08,0.25±0.03,1.03±0.11 and 0.27±0.03,respectively.The above indexes of Vector group were significantly different from those of PRDM5 group(all P<0.05).That 3-Butylphthalidewas bound to PRDM5(free energy-12.55 kcal·mol-1),and binding sites were K413R and D414A.Conclusion 3-butylphthalidebinds to K413R and D414A of PRDM5,inhibiting the function of PRDM5 and improving the apoptosis of neuronal cells induced by oxidative stress.

关 键 词：丁基苯酞 PR结构域蛋白5 氧化应激 神经元细胞 凋亡 

分 类 号：R97[医药卫生—药品]