Research On The Material Basis And Mechanism Of The Anti-anxiety Effect Of Rushes

Anxiety disorder is a neurosis dominated by anxiety,which is mainly divided into panic disorder and generalized anxiety disorder.Anxiety disorders involve repeated episodes of sudden feelings of intense anxiety and fear or terror that reach a peak within minutes.General clinical treatment options for anxiety disorder include psychological intervention,drug intervention and combination therapy,but the commonly used drugs SSRIs(Selective Serotonin Reuptake Inhibitor)and SNRIs(Serotonin-norepinephrine reuptake inhibitors)have an effective rate of 60%-75%.What's worse,there are side effects such as sexual dysfunction,weight gain and sleep disturbance.Therefore,looking for safe anti-anxiety drugs without side effects has become a current research hotspot.Juncus effusus L.often refers to the dried stem pith of Juncus effusus L.,a plant of the genus Juncus in the Juncus family,of which stem pith is being studied a lot these days.However,some studies have confirmed that the phenanthrene content of rush cortex is considerable.It also contains flavonoids and other medicinal ingredients.The basis for the development and utilization of traditional Chinese medicine is the establishment of quality standards and the clarification of pharmacodynamic mechanisms.In order to make full use of the resources,this paper used the whole plant of Juncus effusus L.produced in Fuzhou,Jiangxi,to carry out the following four tasks to lay the foundation for its subsequent development and utilization.(1)Twenty-four mice were randomly divided into three groups,the Juncus effusus L.group(30.0 g(crude medicine)/kg/d),diazepam group(0.3 mg/kg/d)and the control group(0.3 mL/d).The elevated plus maze test was used to evaluate the anti-anxiety efficacy of Juncus effusus L..The evaluation indicators were OT%(the ratio of the time to enter the open arm and the time to enter the closed arm)and OE%(the ratio of the number of times entering the open arm to the number of times entering the closed arm).The larger the ratio,the better the anti-anxiety effect;The percentage of time for mice in the diazepam group to enter the open arm was 49.79±9.14,which was statistically different from that of the blank control group(p<0.001).The OT%of the whole rush grass group was 43.30±15.35,which was also statistically different compared with the blank group(p<0.05),but there was no statistical difference between the diazepam group and the whole rush grass group(p>0.5).There was no significant difference in the percentage of the number of times the three groups entered the open arm(p>0.05);(2)We used Ultra-performance liquid chromatography-Linear Ion Trap Obitriap-Mass Spectrum(UPLC-LTQ-Obitriap-MS)to analyze the chemical composition of the whole plant,then used Ultra-Performance Liquid Chromatography(UPLC)technology for quantitative analysis of the key components:luteolin and dehydroeffusol;A total of 30 compounds were identified,including 11 phenanthrene,8 phenolic acids and 6 flavonoids.The content of luteolin and dehydroeffusol were 0.50± 0.01 mg/g and 1.63± 0.04 mg/g,respectively;(3)After giving 6 rats a high-dose of extracts from Juncus effusus L.,we took serum and brain tissue samples half an hour later.Then we detected the blood and brain components by UPLC-LTQ-Obitriap-MS;A total of 20 blood absorption components were identified,of which 7 were known drug prototype components,and the rest were possible metabolites;a total of 10 brain absorption components were identified,of which 3 were known drug components,and the rest were possible metabolites;(4)The three brain absorption components of Juncus effusus L.:8-carboxy-2-hydroxy-lmethyl-5-vinyl-9,10-dihydrophenanthrene,dehydroeffusol and effusol and anxiety-related protein targets:potassium channels,serotonin transporter,dopamine 2 receptor and yaminobutyric acid receptor were respectively docked,and then molecular dynamics simulations were used to verify the docking results of dehydroeffusol with four protein targets.Through molecular docking,the action modes of three brain components and four protein targets were obtained.Molecular dynamics simulations showed that dehydroeffusol may be hydrogenbonded to potassium channel or transported with serotonin transporter,dopamine 2 receptor through hydrophobic interaction.The elevated plus maze experiment confirmed that 95%Ethanol Extract of 30.0 g(crude medicine)/kg/d administered by gavage to mice for 3 days could improve anxiety-like behaviors of mice.Then we qualitatively analyzed 30 chemical components in Juncus effusus L..The UPLC quantitative method was established for luteolin and dehydroeffusol,and the content of luteolin and dehydroeffusol were 0.50±0.01 mg/g,1.63±0.04 mg/g;Then the blood and brain components of the whole rush grass were determined.The key ingredient dehydroeffusol acted on multiple targets such as potassium channel protein,serotonin transporter and dopamine 2 receptor protein to exert anti-anxiety effects after entering the brain through the blood-brain barrier.