海南医学院学报2023，Vol.29Issue(24)：1855-1862,1869,9.DOI:10.13210/j.cnki.jhmu.20231011.004
基于分子对接、分子动力学和体内实验研究毛蕊花苷对帕金森病小鼠的作用机制
Mechanism of action of verbascoside in mice with Parkinson's disease based on molecular docking,molecular dynamics and in vivo experiments
摘要
Abstract
Objective:To explore the improving effect of verbascoside on Parkinson's disease(PD)model mice and to clar-ify the possible mechanism.Methods:60 C57BL/6 male mice were randomly divided into normal group,model group,low,mid-dle and high dose groups(30,60,120 mg/kg,respectively).After a week of adaptive culture,except for the blank group,the subacute model of PD was established by intraperitoneal injection of 1-methyl-4-phenyl-1(MPTP)in each group for 5 consecutive days.After a week of adaptive culture,except for the blank group,the subacute model of PD was established by intraperitoneal in-jection of 1-methyl-4-phenyl-1(MPTP)in each group for 5 consecutive days.The establishment of the model,the drug group was intragastrically administered continuously for 7 days,and the mice in the normal group were injected with the same amount of sa-line.The behavioral changes of mice were observed by open field test,pole climbing test and grip test;the morphology,apoptosis and Nissl bodies of substantia nigra neurons were observed by HE staining and Nissl staining;the ultrastructural changes of neu-rons were observed by transmission electron microscope;the positive expressions of tyrosine hydroxylase(TH),α-synuclein(α-Syn)and toll-like receptor 4(TLR4)were detected by immunohistochemistry.The protein levels of TH,α-Syn,TLR4,NF-κB and P-NF-κB in substantia nigra of mice were detected by Western blot,and the levels of inflammatory factors IL-1 β,IL-6 and tu-mor necrosis factor-α(TNF-α)in serum were detected by Elisa.Molecular docking was used to verify the binding ability of ver-bascoside to TLR4/NF-κB signal pathway and related factors,and molecular dynamics was used to verify the binding mode and stability of verbascoside with compounds.Results:Compared with the normal group,the total walking distance,average speed,free activity time,pole climbing time and forelimb grip in the model group were significantly longer than those in the normal group(P<0.05).The ultrastructural changes of neurons,nuclear lysis,deformation and mitochondrial damage were observed under transmission electron microscope,and the number and morphological changes of HE staining substantia nigra neurons were de-creased(P<0.05).Nissl staining showed that the number of Nissl positive cells decreased(P<0.05),the positive expression of TH decreased and the positive expression of α-syn and TLR4 increased(P<0.05).Western blotting showed that the expression of TH protein decreased,the expression of α-syn,TLR4,NF-κB p65 protein increased(P<0.05),and the expression of IL-1β,IL-6 and TNF-α in serum increased significantly(P<0.05).Compared with the model group,the activity ability of mice in the treat-ment group was significantly improved(P<0.05),the morphology of neurons gradually recovered and the number of neurons in-creased(P<0.05).The number of Nissl positive cells increased(P<0.05).The expression of TH protein increased,while the expression of α-syn,TLR4,NF-κB p65 protein decreased(P<0.05),and the expression of IL-1β,IL-6,TNF-α in serum de-creased significantly(P<0.05).The results of molecular docking and molecular dynamics simulation show that the combination of mulberry glycoside with the compound is good and the property is stable.Conclusion:Verbascoside has protective effect on PD mice,and its mechanism may be related to TLR4/NF-κ B pathway,and then regulate neuroimmunity.关键词
帕金森病/毛蕊花苷/Toll样受体4/分子对接Key words
Parkinson's disease/Verbascoside/Toll-like receptor 4/Molecular docking分类
医药卫生引用本文复制引用
阿迪莱·艾比布力,克力比奴儿·赛地尔丁,逯冉冉,杨新玲..基于分子对接、分子动力学和体内实验研究毛蕊花苷对帕金森病小鼠的作用机制[J].海南医学院学报,2023,29(24):1855-1862,1869,9.基金项目
This study was supported by Special Funds for LOCAL SCIENce and Technology Development Guided by the Central Government(ZYD2022C17) （ZYD2022C17）
National Natural Science Foundation of China(82160232) 中央引导地方科技发展专项资金项目(ZYD2022C17) （82160232）
国家自然科学基金(82160232) （82160232）
