Application of poria cocos water decoction in preparation of anxiolytic drugs
阅读说明：本技术 茯苓水煎液在制备抗焦虑药物中的应用 (Application of poria cocos water decoction in preparation of anxiolytic drugs ) 是由 崔瑛 李瑞洁 崔璨 李玲玲 史银玥 楚玉玺 于 2020-07-29 设计创作，主要内容包括：本发明涉及茯苓水煎液在制备抗焦虑药物中的应用,可有效解决制备抗焦虑药物,开拓茯苓药物新用途的问题,其解决的技术方案是,所述的茯苓水煎液是,取茯苓饮片,每次加10倍重量体积的蒸馏水100℃加热回流2次,每次1h,合并两次滤液,4000r·min<Sup>-1</Sup>离心5min,浓缩成相当于含生药0.5-1.5g/mL的茯苓水煎液。本发明原料丰富,易生产制备,所制备的水煎液具有抗焦虑之功效,有效用于制备抗焦虑药物,开拓了茯苓的新药用价值,是抗焦虑药物上的创新,有显著的经济和社会效益。(The invention relates to application of a poria cocos water decoction in preparation of an anxiolytic drug, which can effectively solve the problems of preparation of the anxiolytic drug and development of new application of the poria cocos drug and solves the technical scheme that the poria cocos water decoction is prepared by adding 10 times of distilled water by weight and volume at 100 ℃ into poria cocos decoction pieces, heating and refluxing for 2 times and 1 hour each time, combining two filtrates, and mixing the filtrates at 4000 r.min ‑1 Centrifuging for 5min, and concentrating to obtain Poria water decoction containing crude drug 0.5-1.5 g/mL. The invention has rich raw materials and easy production and preparation, and the prepared water decoction has the effect of antianxiety, is effectively used for preparing the antianxiety medicament, develops the new medicinal value of the tuckahoe, is innovation on the antianxiety medicament and has obvious economic and social benefits.)
1. The application of a Poria water decoction in preparing anxiolytic drugs is prepared by adding 10 times of distilled water by weight into Poria decoction pieces, heating and refluxing for 2 times and 1h each time, soaking for 1h during the first heating and refluxing, mixing the filtrates, and heating for 4000 r.min-1Centrifuging for 5min, and concentrating to obtain Poria water extractive solution containing crude drug 0.5-1.5g/mL, wherein the weight volume is solid in g and liquid in mL.
2. The use of the aqueous poria cocos decoction of claim 1 for the preparation of a medicament for improving the symptoms of anxiety disorders, increasing the GABA content in the brain, and decreasing the GLU and 5-HT and DA content.
Technical Field
The invention relates to medicine, in particular to application of tuckahoe water decoction in preparing an anxiolytic drug.
Background
Poria is Poria cocos (Schw.) wolf of PolyporaceaeWolfiporia cocos(Schw.) Wolf's dried sclerotium, as listed above, was recorded in Shen nong's herbal Jing.
Poria cocos is thought to have a better effect of calming heart and tranquilizing mind throughout the past, as in Yun of Shen nong Ben Cao Jing: "Fu Ling … … governs adverse qi in chest and hypochondrium, worry anger, fright evil, palpitation, and pain in heart lower node … …. Long-term administration of Anhui spirit and tranquilization". The famous physicians' book says that it is effective in nourishing yin, invigorating qi and keeping spirit. In the statement of the property of herbs, "it is good at tranquilizing mind". The Ben Cao Bei Yao (herbal treatise) points out that tuckahoe can 'quiet heart and tonify qi, regulate ying and regulating physical defense, and calm soul'. The book Ben Cao Chong Yuan considers that tuckahoe is taken for a long time to calm the spirit of liver and nourish the heart. In the chapter of De Lu with Ben Cao, it is said that it can induce heart-fright heat, so it can treat fright. The book Ben Cao fen Jing (materia Medica division of records) says: poria cocos, Poria cocos. The white one enters qi system, and benefits spleen and calms heart. "Zhongzhuixi Lu" in Zhongzhao Zhong Can xi di.
Pharmacological experiments show that the tuckahoe water decoction has the central inhibition effect of cooperating with the pentobarbital sodium, can obviously prolong the sleep time of mice, and shows that tuckahoe has the sedative and hypnotic effects. The carboxymethyl pachymaran injection can enhance the inhibition of thiopentone sodium on the center of a mouse, and obviously prolong the lasting time of disappearance of righting reflex of the mouse. However, there is no report on the anxiolytic effect of tuckahoe decoction.
The anxiety neurosis can be seen in various traditional Chinese medicine syndrome types, such as liver depression and spleen deficiency type, heart-spleen deficiency type, yin deficiency and fire excess type and the like, and the poria cocos has a good spleen tonifying effect.
Disclosure of Invention
In view of the above situation, in order to overcome the defects of the prior art, the present invention aims to provide an application of a poria cocos water decoction in preparation of an anti-anxiety drug, which can effectively solve the problems of preparation of the anti-anxiety drug and development of new uses of the poria cocos drug.
The technical scheme for solving the problem is that tuckahoe water decoction is used for preparing the anxiolyticThe application of the Chinese medicinal composition comprises the steps of taking poria cocos decoction pieces, adding distilled water with the weight and volume being 10 times of that of the poria cocos decoction pieces each time, heating and refluxing for 2 times at 100 ℃ and 1h each time, soaking for 1h before first extraction, combining filtrates of the two times, and heating for 4000 r.min-1Centrifuging for 5min, and concentrating to obtain Poria water decoction containing crude drug 0.5-1.5g/mL, wherein the weight volume is solid in g and liquid in mL.
The invention has rich raw materials and easy production and preparation, and the prepared water decoction has the effect of antianxiety, is effectively used for preparing the antianxiety medicament, develops the new medicinal value of the tuckahoe, is innovation on the antianxiety medicament and has obvious economic and social benefits.
Detailed Description
The following detailed description of the embodiments of the present invention refers to the accompanying drawings.
In the specific implementation of the invention, the application of the tuckahoe water decoction in preparing the anxiolytic drug selects Anhui Anqing tuckahoe decoction pieces purchased from the third subsidiary hospital of Henan Chinese medicine university, and the Anhui Anqing tuckahoe decoction pieces are identified as the tuckahoe which is a fungus of the family Polyporaceae by Chen Ganzhizu of Henan Chinese medicine universityWolfiporia cocos(Schw.) Wolf drying sclerotium, soaking Poria decoction pieces in 10 times of distilled water for 1 hr, heating and refluxing at 100 deg.C for 1 hr, and filtering to obtain first filtrate; adding distilled water 10 times the weight and volume of the original medicine into the medicine residue, heating and refluxing for 1h, and filtering to obtain a second filtrate; mixing the two filtrates, mixing, and stirring at 4000r min-1Centrifuging for 5min, concentrating to obtain Poria water decoction containing crude drug 0.5-1.5g/mL, and making into oral dosage of 5 g/kg-1、10g·kg-1、15g·kg-1The water decoction of tuckahoe.
The invention has rich raw materials, easy production and preparation, convenient taking and good effect, the tuckahoe water decoction has the effect of antianxiety, is effectively used for preparing antianxiety drugs, exploits the new medicinal value of tuckahoe, and obtains very good beneficial technical effect through experiments, and the related experimental data are as follows:
first, the influence of Poria cocos water decoction on mouse anxiety model
1 materials of the experiment
1.1 Experimental animals
The SPF-level healthy male Kunming-breed mice weigh 18-22 g, are purchased from the experimental animal center of Henan province, and have a license number: SCXK (yu) 2017-: no. dw2019060015. The mice are raised in an independent air supply isolation cage of IVC-II mice of animal experiment center of Chinese medicine university in Henan, the temperature of a laboratory is controlled at 22 +/-2 ℃, and the use license number SYXK (Yu) 2015 and 0005 of an experimental unit are used.
1.2 Experimental drugs
The invention relates to a tuckahoe water decoction.
Diazepam, purchased from san fu hospital, university of traditional Chinese medicine, south of Henan, manufacturers: tianjingsheng pharmaceutical products ltd, specifications: 2.5 mg/tablet × 100 tablets, approved literature: national drug standard H12020119, production lot number: 1806008.
mouse GABA, GLU, 5-HT, DA kits (Shanghai Elisa Biotech Co., Ltd.), batch No.: E20190801A.
1.3 Experimental instrumentation
Rat IVC independent air supply isolation cage (model: IVC-II, Suzhou von Willebrand laboratory animal facilities Co., Ltd.);
PM-200 elevated cross labyrinth device: chengduta alliance technologies, Inc.;
mouse light and dark box devices were purchased from shanghai soft information technology ltd;
microplate readers (type: Epoch) were purchased from BioTeK Instruments, Inc.
2 protocol
2.1 preparation of the drug
2.1.1 Poria Water decoction the Poria water decoction prepared by the invention is prepared into a concentration of 0.5 g/mL-1、1.0g·mL-1、1.5g·mL-1The liquid medicine of (1) is ready for use.
2.1.2 preparation of diazepam suspension diazepam tablet is ground, dissolved in distilled water, treated by ultrasonic treatment, and prepared into 0.25 mg/mL-1The diazepam suspension is ready for use and is fully shaken up before use.
2.2 grouping and administration
SPF grade healthy male Kunming species72 mice with the weight of 18-22 g are labeled and weighed after being adaptively fed for 3 days, and are randomly divided into 6 groups, namely a blank control group (not subjected to an ethological experiment), an anxiety model group, a positive control group, a high-dose, medium-dose and low-dose poria cocos water decoction group, and 10 mice in each group. Mice were screened and grouped in open-box experiments 1 day prior to dosing to ensure that there was no significant difference in the behavioral abilities of the groups of mice. The administration is carried out by gavage every morning for 5 days, and the administration volume is 0.1 mL-10 g-1. The blank control group and the anxiety model group are administered with distilled water by intragastric administration every day, and the positive control group is administered with 0.0025 g.kg by intragastric administration every day-1The diazepam suspension is prepared by respectively intragastrically administering 5 g/kg of Poria with high, medium and low dosage-1、10g·kg-1、15g·kg-1The water decoction of tuckahoe. The behavioural test was performed 1h after the last gavage.
2.3 behavioural testing
2.3.1 elevated plus maze experiment Each mouse was placed in a separate mouse cage for 5min before starting the experiment, and then placed on the open platform in the center of the elevated plus maze with the head facing the open arm and allowed to freely explore. And (4) tracking and recording by using a computer, and collecting the movement data of the mouse on the elevated plus maze within 5 min. OE%, i.e. the number of times of entry of open arm/(the number of times of entry of open arm + the number of times of entry of closed arm) and OT%, i.e. the open arm retention time/(the open arm retention time + the arm retention time) were used as the index for evaluating the anxiolytic effect of the Poria cocos aqueous decoction. After the test of each mouse is finished, excrement such as feces is cleared in time, the maze is sprayed and wiped by low-concentration alcohol if necessary to remove peculiar smell, and then the next test is carried out. In the whole experiment process, the experimental environment is kept quiet, the experimental device is clean, and the adverse effect of the external environment on the experimental result is avoided as much as possible.
2.3.2 light and dark box experiment after the elevated plus maze experiment is finished, immediately carrying out a light and dark box experiment, putting a mouse into the center of the light box in a posture of back to the dark box, and observing the shuttling times between the light box and the dark box within 10min of the mouse, wherein the shuttling times are used as indexes for evaluating the anxiolytic effect of the poria cocos water decoction. After the test of each mouse is finished, excrement such as excrement is removed in time, the bottom of the box is sprayed and wiped by low-concentration alcohol if necessary to remove peculiar smell, and then the next test is carried out. In the whole experiment process, the experimental environment is kept quiet, the experimental device is clean, and the adverse effect of the external environment on the experimental result is avoided as much as possible.
2.4 correlation index detection
Immediately after the behavioral experiment, the mouse is killed by removing cervical vertebra, the head is quickly cut off, the whole brain is quickly peeled off on an ice bench, the brain is precisely weighed by a ten-thousandth balance, the normal saline is added according to the proportion of brain tissue (g) to the normal saline (mL) =1:4 to prepare homogenate, and the homogenate is processed at 4 ℃ and 3500 r.min-1Centrifuging for 20min, collecting supernatant to obtain mouse brain tissue homogenate, and storing in-80 deg.C refrigerator. The contents of GABA, GLU, 5-HT and DA in the brain tissue of the mouse are determined by an enzyme-linked immunosorbent assay strictly according to the instruction of a kit.
2.5 statistical treatment
Experimental data were analyzed for one-way anova using SPSS23.0 software, and mean ± standard deviation (for each group of data) (SPSS 23.0 software:)
3 results
3.1 Effect of Poria cocos Water decoction on anxiety behavior in mice
After administration, the positive control group, the poria water decoction is low, the OE% and OT% of the mice in the middle dose group in the EPM device and the shuttle times in the light and dark boxes are all obviously increased compared with the anxiety model group (the shuttle times in the positive control group, the poria water decoction and the middle dose group are all obviously increased by the administration method of the composition: (the anxiety model group is notP＜0.05，P< 0.01), the OT% of the mice in the tuckahoe water decoction high-dose group is obviously increased in the EPM device (P< 0.01). The above experimental results show that 5 g.kg-1~15 g·kg-1The tuckahoe decoction has certain intervention effect on anxiety behaviors of mice in an EPM and light and dark box device, and the tuckahoe decoction is prompted to have certain anxiolytic effect on the anxiety mice. See table 1.
TABLE 1 Effect of Poria cocos decoction on anxiety behavior in mice: ( ±s，n=10）
Group of
Dosage (g.kg)-1）
OE%（%）
OT%（%）
Number of shutdowns (times)
Anxiety model group
—
47.60±2.95
44.45±5.56
25.80±5.03
Positive control group
2.5×10-3
56.70±5.23**
57.03±5.57**
42.90±8.27**
Poria cocos water decoction low dose group
5.0
52.40±2.50*
54.88±5.11**
32.30±6.41*
Medium dosage of Poria cocos decoction
10.0
55.00±4.00**
50.14±3.45*
32.60±7.06*
Poria cocos water decoction high dose group
15.0
50.70±3.06
51.03±4.09**
29.00±3.68
Note: in comparison to the set of anxiety models,* P＜0.05，** P＜0.01
3.2 Effect of Poria cocos Water decoction on the content of GABA, GLU, 5-HT and DA in mouse anxiety model brain tissue
Significantly reduced GABA content in brain tissue of anxiety model mice compared to placebo group (P<0.01), the content of GLU and 5-HT is significantly increased (P<0.05，P<0.01), the DA content tends to increase; after administration, the GABA content in the brain tissue of mice in the positive control group and the tuckahoe water decoction is obviously increased compared with that in the anxiety model group (the GABA content in the brain tissue of mice in the low, medium and high dose groups of the tuckahoe water decoction is obviously increased: (P<0.01), the content of GLU, 5-HT and DA is all reduced significantly: (P<0.05，P<0.01). The above experimental results show that 5 g.kg-1~15 g·kg-1The poria cocos water decoction can effectively improve the abnormal change trend of the GABA, GLU, 5-HT and DA contents of brain tissues of anxiety mice, is consistent with experimental results of behaviourology, and is probably a mechanism for playing an anxiolytic role.
TABLE 2 influence of Poria decoction on GABA, GLU, 5-HT, DA content in mouse brain tissue: (
Group of
Dosage (g.kg)-1)
GABA(μmol·L-1)
GLU(mg·L-1)
5-HT(ng·mL-1)
DA(pg·mL-1)
Blank control group
—
33.25±3.24
214.45±20.50
777.22±38.13
386.35±17.86
Anxiety model group
—
28.09±1.91##
245.01±13.44##
817.09±57.38#
397.56±12.30
Positive control group
2.5×10-3
31.26±1.71**
210.25±11.34**
775.63±43.40*
359.77±15.56**
Poria cocos water decoction low dose group
5.0
32.40±2.07**
233.01±9.26*
771.20±41.50*
371.41±21.44**
Medium dosage of Poria cocos decoction
10.0
32.54±3.10**
233.64±6.84*
738.61±25.75**
373.99±14.94**
Poria cocos water decoction high dose group
15.0
31.75±2.43**
216.99±7.03**
748.73±36.37**
378.74±12.06*
Note: in comparison to the set of anxiety models,# P＜0.05，## P＜0.01。
4. small knot
The experiment observes that after continuously administering the tuckahoe water decoction for 5 days, the OE% and OT% values of mice in an EPM device can be obviously improved, and the shuttling times in a light box and a dark box are obviously increased, which shows that the tuckahoe water decoction has an anxiolytic effect on mice with anxiety in the device. The mechanism of anxiolytic effect may be achieved by improving central amino acid neurotransmitter levels, such as increasing GABA content and decreasing GLU content, and improving monoamine neurotransmitter levels, such as inhibiting 5-HT and DA levels. This result is similar to the effect produced by diazepam, and has preliminarily confirmed its potential as an anxiolytic.
Effect of Poria cocos water decoction on spleen deficiency anxiety rats
1 materials of the experiment
1.1 Experimental animals
The male S SD rat is SPF-grade and 180-220 g in weight, and is purchased from Jinan Pengyue experimental animal breeding company Limited in Shandong province, with a license number: SCXK (Lu) 2019-. The rat feed is purchased from experimental animals center in Henan province, and the batch number is as follows: 41003100006603.
1.2 Experimental drugs
The invention relates to a tuckahoe water decoction.
Folium sennae, radix Aconiti lateralis Preparata, cortex Cinnamomi, and fructus evodiae decoction pieces, all purchased from the third subsidiary hospital of Henan university of traditional Chinese medicine, and identified as Cassia angustifolia of Leguminosae respectively by Chen following Qing and Dai of Henan university of traditional Chinese medicineCassia angustifoliaDried leaflets of Vahl, Aconitum carmichaeli Debx of RanunculaceaeAconitum carmichaeliiProcessed product of seed root of Debx, and Cinnamomum cassia Presl of LauraceaeCinnamomum cassiaDried bark of Presl, Rutaceae plant evodia rutaecarpaEuodia rutaecarpa(Juss.) dried near ripe fruit of Benth.
Diazepam, purchased from san fu hospital, university of traditional Chinese medicine, south of Henan, manufacturers: tianjingsheng pharmaceutical products ltd, specifications: 2.5 mg/tablet × 100 tablets, approved literature: national drug standard H12020119, production lot number: 1806008.
rat GABA, GLU, GABA-T, GAD, GS kits (Shanghai Elisasa Biotech Co., Ltd.), batch No. E20191101A
1.3 Experimental instruments
Rat IVC independent air supply isolation cage: suzhou von experimental animal facilities limited);
PM-200 elevated cross labyrinth device: chengduta alliance technologies, Inc.;
mouse light and shade case device: shanghai Xin soft information technology, Inc.;
microplate readers (type: Epoch) were purchased from BioTeK Instruments, Inc.
2 protocol
2.1 preparation of the drug
2.1.1 preparation of Poria Water decoction prepared by the invention is prepared into Poria Water decoction with concentration of 0.4 g.mL-1、0.8g·mL-1、1.2g·mL-1The liquid medicine of (1) is ready for use.
2.1.2 preparation of Cassia angustifolia decoction pieces, soaking in 10 times of distilled water for 1h, heating and refluxing for 2 times, 1h each time, mixing the filtrates, 4000 r.min-1Centrifuging for 5min, concentrating, and making into 1 g/mL-1The senna leaf decoction.
2.1.3 preparation of Mixed decoction of radix Aconiti lateralis, cortex Cinnamomi and fructus evodiae by soaking radix Aconiti lateralis, cortex Cinnamomi and fructus evodiae in 10 times of distilled water for 1 hr, heating and refluxing for 2 times (1 hr each time), mixing the filtrates, and heating for 4000 r.min-1Centrifuging for 5min, concentrating, and making into 1 g/mL-1The decoction of (1).
2.1.4 preparation of diazepam suspension diazepam tablet is ground, dissolved in distilled water, treated by ultrasonic treatment, and prepared into 0.25 mg/mL-1The diazepam suspension is ready for use and is fully shaken up before use.
2.2 grouping and administration
The preparation of the model with spleen yang deficiency and spleen yin deficiency is divided into two stages, wherein the first stage adopts a method of improper diet combined with overstrain to cause a spleen qi deficiency model, and the second stage adopts bitter cold drugs or yin-impairing drugs to respectively cause a spleen yang deficiency model and a spleen yin deficiency model. In the experiment, except for the blank control group and the anxiety model group, the rats in the other groups were subjected to a weight-bearing swimming experiment (so that the rats in the other groups are subjected to the weight-bearing swimming experiment) every morningPutting a transparent barrel with the height of 70 cm and the diameter of 15 cm into a rat at one time for swimming, loading a weight accounting for 5% of the weight of the rat, taking out the rat after 10min to exhaustion by taking the rat as the standard that the rat cannot float upwards after the nose is 10 s, and wiping water with a towel) and continuously carrying out 22 d. From day 16, on the basis of swimming with heavy load, the spleen yang deficiency anxiety model group and the high, medium and low dose administration groups were administered with 10 g/kg per day in the afternoon-1The senna leaf decoction; spleen yin deficiency anxiety model group and high, medium and low dose administration group are intragastrically administered with 10 g/kg every day in the afternoon-1The water decoction of aconite, cinnamon and evodia rutaecarpa is continued for 7 days. During the molding period, rats are fed with food for 1 d and fasted for 2 d in a circulating way, and the drinking water of each group is kept normal. Experiment 23d, positive group gavage diazepam, each administration group of Poria cocos is gavage with Poria cocos water decoction of corresponding dose, blank control group, anxiety model group, spleen yang deficiency anxiety model group and spleen yin deficiency anxiety model group are gavage with distilled water, and the continuous 7 d. Experiment 29 d, except for the placebo group, the rats in each group were made anxiety models on an anxiety device and were observed for anxiety behavior simultaneously.
2.3 behavioural testing
The elevated plus maze experiment and the light and dark box experiment are the same as the methods under the item of '2.3 behavioural tests' in the previous section.
2.4 correlation index detection
The detection is carried out by the method under the item of '2.4 related indexes detection' in the previous part.
2.5 statistical treatment
Experimental data were analyzed for one-way anova using SPSS23.0 software, and mean ± standard deviation (for each group of data) (SPSS 23.0 software:)S) is represented byP<0.05 indicates that there is a significant difference,P<0.01 indicates a very significant difference.
3 results
3.1 Effect of Poria cocos Water decoction on spleen Yang deficiency anxiety rats
3.1.1 Effect of Poria cocos Water decoction on general status of rats with anxiety due to spleen Yang deficiency
Compared with a blank control group, the molded rats generally have the phenomena of weight loss, dry hair, difficult grabbing, perianal and buttock filth, loose stool, obvious reduction of swimming endurance and the like, and the phenomena are consistent with the clinical manifestations of spleen-yang deficiency. After the model building is finished and the administration is started, the weight of rats in each dose group of tuckahoe water decoction is obviously increased, the hair is glossy, the symptoms such as loose stool and the like are improved to different degrees.
Beginning from the 10 th model making, the anal temperature of the rats of each model making group is gradually reduced compared with that of the anxiety model group; (19 d) starting from the model creation, the anal temperature of the rats in each model creation group was significantly reduced compared to the anxiety model group: (P<0.01) which trend continues until the time of molding 22 d and the administration after molding. The reduction of the anal temperature of the rat accords with the characteristics of the spleen-yang deficiency syndrome of the traditional Chinese medicine syndrome. After the model building is finished and the administration is started, the anal temperature of rats in each dose group of tuckahoe decoction solution begins to rise gradually, and the high dose group is obviously increased compared with the spleen yang deficiency anxiety model group (P<0.05), which shows that the tuckahoe water decoction has certain improvement effect on the reduction of the anal temperature of rats with spleen yang deficiency.
3.1.2 Effect of Poria cocos Water decoction on anxiety behavior of rats with spleen yang deficiency
Compared with the anxiety model group, the spleen yang deficiency anxiety model group rats have a significant reduction of OE% in the EPM device (P<0.05), the number of shuttles in OT% and the light and dark boxes both tended to decrease, indicating that the deficiency of spleen yang would aggravate the anxiety behavior of the rats to some extent. After the administration treatment, compared with the spleen yang deficiency anxiety model group, the OE% of rats in the positive control group, the tuckahoe water decoction and the high dose group in the EPM device is obviously increased (P<0.05，P<0.01); significant increase in OT% in the positive control rat EPM device (a) ((b))P<0.01), the OT% of the tuckahoe water decoction is obviously increased in low, medium and high dose groups; the shuttle times in the light and dark boxes of rats in the positive control group and the tuckahoe water decoction high-dose group are obviously increased (P<0.05，P<0.01). The above experimental results show that 4 g.kg-1~12 g·kg-1The decoction of Poria can improve anxiety behavior of spleen yang deficiency anxiety rat in EPM device and light and dark box. See table 3.
TABLE 3Influence of Poria cocos aqueous extract on anxiety behavior of rats with spleen yang deficiency (±s，n=10）
Group of
Dosage (g.kg)-1）
OE%（%）
OT%(%)
Number of shutdowns (times)
Anxiety model group
—
27.99±13.29
12.65±10.70
4.40±3.66
Spleen yang deficiency anxiety model group
—
17.60±11.42#
5.60±4.14
4.00±2.40
Positive control group
2.0×10-3
63.06±5.42**
31.01±18.34**
8.50±3.24**
Poria cocos water decoction low dose group
4.0
29.32±8.02*
11.16±13.22
6.10±3.90
Medium dosage of Poria cocos decoction
8.0
33.17±10.09**
16.26±8.78
6.30±3.23
Poria cocos water decoction high dose group
12.0
37.81±14.17**
12.14±11.74
7.60±4.06*
Note: in comparison to the set of anxiety models,# P＜0.05，## Pless than 0.01, compared with the spleen yang deficiency anxiety model group,* P＜0.05，** P＜0.01
3.1.3 Effect of Poria cocos Water decoction on transmitter content in Hippocampus tissue of rats with anxiety due to spleen yang deficiency
Compared with the blank group by photography, the GABA content in the hippocampal tissues of the rats in the anxiety model group and the spleen yang deficiency anxiety model group is obviously reduced (P< 0.05), the GAD, GS activity was significantly reduced (P< 0.05), the GABA-T activity was significantly increased: (P< 0.05). After administration, compared with the spleen yang deficiency anxiety model group, the positive control group and the tuckahoe water decoction low, medium and high dose groups can significantly reduce the great reductionGLU content and GABA-T Activity in Hippocampus tissues of mice: (P<0.05，P<0.01); the positive control group, the tuckahoe water decoction medium and the high dose group can obviously increase the GABA content in the hippocampus tissues (the)P<0.05); the positive control group and the tuckahoe water decoction low dose group can obviously enhance the activity of GAD in rat hippocampus tissues (the activity of GAD in rat hippocampus tissue)P<0.01); the tuckahoe water decoction high-dose group can obviously enhance the GS activity in rat hippocampal tissues (P<0.01). The above experimental results show that 4 g.kg-1~12 g·kg-1The tuckahoe decoction has certain reversion effect on GABA content, GAD and GS activity reduction, GLU content and GABA-T activity increase of rats with spleen yang deficiency anxiety. Suggesting that the action mechanism of tuckahoe for resisting anxiety is probably related to the regulation of enzyme activities of GAD, GABA-T and GS in hippocampal tissues of rats with anxiety caused by spleen yang deficiency, thereby regulating the dynamic balance of GLU and GABA content. See tables 4-5.
TABLE 4 influence of Poria decoction on GABA and GLU content in hippocampal tissue of anxiety rat due to spleen yang deficiency: (±s，n=10）
Group of
Dosage (g.kg)-1）
GABA(μmol·L-1)
GLU(mg·L-1)
Blank control group
—
42.66±3.77
205.24±16.66
Anxiety model group
—
38.66±4.34△
224.52±21.18△
Spleen yang deficiency anxiety model group
—
36.63±3.25△
222.31±15.00△
Positive control group
2.0×10-3
41.38±4.26*
201.31±14.66*
Poria cocos water decoction low dose group
4.0
36.27±5.05
209.74±18.94
Medium dosage of Poria cocos decoction
8.0
40.82±4.58*
191.27±19.72**
Poria cocos water decoction high dose group
12.0
40.63±2.96*
202.99±31.11*
Note: compared with the blank control group, the composition of the composition,△ P＜0.01，△△ Pless than 0.05; compared with the spleen yang deficiency anxiety model group,* P＜0.05，** P＜0.01
TABLE 5 influence of Poria decoction on GABA-T, GAD and GS content in hippocampal tissue of anxiety rat with spleen yang deficiency: (
Group of
Dosage (g.kg)-1）
GAD（U·L-1）
GABA-T（U·L-1）
GS（U·L-1）
Blank control group
—
238.37±29.83
192.00±9.92
472.41±16.81
Anxiety model group
—
204.77±25.18△
208.76±10.94△
431.20±31.30△△
Spleen yang deficiency anxiety model group
—
195.83±21.97△
204.49±9.64
404.38±67.58△△
Positive control group
2.0×10-3
237.87±44.82**
185.6±14.87**
411.13±36.11
Poria cocos water decoction low dose group
4.0
214.71±26.11
191.67±9.96*
388.32±13.47
Medium dosage of Poria cocos decoction
8.0
225.61±12.84
188.13±20.24**
409.67±43.30
Medium dosage of Poria cocos decoction
8.0
234.83±27.95*
185.68±12.7**
430.88±42.82
Note: compared with the blank control group, the composition of the composition,△P＜0.01，△△p is less than 0.05; p < 0.05, P < 0.01, compared to the spleen yang deficiency anxiety model group
3.2 Effect of Poria cocos Water decoction on rats with anxiety due to spleen-yin deficiency
3.2.1 Effect of Poria cocos Water decoction on general State of rats with anxiety due to spleen-yin deficiency
Compared with a blank control group, the rats after the model building generally have the phenomena of restlessness, irritability, dry stool, weight loss, decline of swimming endurance, high anal temperature rise and the like. After administration of each administration group, the weight of rats is increased, the anal temperature is reduced, the stool form is recovered to be normal, and symptoms such as restlessness and the like are relieved.
Starting from the 3d model building, the anal temperature of the rats of each model building group is gradually increased compared with that of the anxiety model group; (19 d) starting from model 19 d, the anal temperature of the rats in the model group with anxiety due to deficiency of spleen-yin is remarkably increased compared with that in the model group with anxiety: (P<0.05，P<0.01); the phenomenon of anal temperature rise of rats of each modeling group is continued until the modeling is finished and the drug administration process after the modeling is finished. The increase of the anal temperature of the rat accords with the characteristics of spleen-yin deficiency syndrome of traditional Chinese medicine syndrome. After administration, the rising trend of the anal temperature of rats in each dose group of tuckahoe decoction is relieved, wherein the anal temperature of rats in a high dose group is obviously reduced in D24 days compared with that in a spleen yin deficiency anxiety model group (the dose is that the anal temperature of rats in a high dose group is obviously reduced by the administration of tuckahoe decoction, the medicine is not used for treating chronic hepatitis BP<0.05). The results show that the tuckahoe water decoction has a certain improvement effect on the increase of the anal temperature of rats with anxiety due to spleen-yin deficiency.
3.2.2 Effect of Poria cocos Water decoction on anxiety behavior of rats with deficiency of spleen-yin
Compared with the anxiety model group, the numbers of shuttles in the light box and the dark box of OE% and OT% in the rat EPM device of the spleen-yin deficiency anxiety model group are reduced, and the anxiety behaviors of the rat are aggravated to a certain extent; after administration, compared with the spleen yin deficiency anxiety model group, the OE% of rats in the positive control group, the tuckahoe water decoction low, medium and high dose groups in the EPM device is obviously increased (OE%P<0.05，P<0.01); OT% of rats in positive control group and poria cocos water decoction high-dose group in EPM deviceRemarkably increase (P<0.05); the shuttling times of rats in the positive control group, the low tuckahoe water decoction and the high dose group in the light and dark box are obviously increased (P<0.01). The above experimental results show that 4 g.kg-1~12 g·kg-1The Poria decoction can improve anxiety behavior of rats with anxiety due to spleen-yin deficiency in EPM and light and dark box. As in table 6.
TABLE 6 influence of Poria decoction on spleen-yin deficiency in rat EPM and light and dark box experiment: (
Group of
Dosage (g.kg)-1）
OE%（%）
OT%(%)
Number of shutdowns (times)
Anxiety model group
—
27.99±13.29
12.65±10.70
4.40±3.66
Spleen yin deficiency anxiety model group
—
24.19±12.40
9.09±9.28
3.00±2.10
Positive control group
2.0×10-3
59.40±10.31**
30.97±25.40*
8.30±2.23**
Poria cocos water decoction low dose group
4.0
42.39±12.40**
15.87±16.96
9.40±3.86**
Medium dosage of Poria cocos decoction
8.0
45.17±14.36**
18.32±14.50
5.50±1.72
Poria cocos water decoction high dose group
12.0
55.46±12.96**
29.25±14.15*
7.90±2.24**
Note: compared with the spleen yin deficiency anxiety model group:* P＜0.05，** P＜0.01
3.2.3 Effect of Poria cocos Water decoction on transmitter content in Hippocampus tissue of rats with anxiety due to spleen-yin deficiency
Comparison with blank group for photographs, anxiety model group rat hippocampal groupSignificant reduction in GABA content in tissue (P< 0.05), a significant reduction in GAD and GS activity (P<0.01); (the content of GABA in hippocampal tissues of rats in the spleen-yin deficiency anxiety model group is remarkably reduced compared with that in the anxiety model group: (P< 0.05). After administration, the GABA content in hippocampal tissue of rats in the positive control group and the dose group in the tuckahoe water decoction is remarkably increased compared with the spleen yin deficiency anxiety model group (P<0.05，P<0.01); the GABA content in the hippocampus tissues of rats in the positive control group and the tuckahoe water decoction low, medium and high dose groups is obviously increased (P<0.05，P<0.01); the activity of GAD in hippocampal tissues of rats in a low-dose group and a positive control group of tuckahoe decoction is remarkably improved (P<0.05，P<0.01); significant reduction of GABA-T activity in hippocampal tissues of rats in low and medium dose tuckahoe decoction and positive control group (P<0.05，P<0.01), the activity of GS in rat hippocampus tissues of the dose group in the tuckahoe water decoction is obviously improved (P<0.01). The above experimental results show that 4 g.kg-1~12 g·kg-1The tuckahoe decoction has certain reversion effect on GABA content, GAD and GS activity reduction, GLU content and GABA-T activity increase in hippocampal tissues of rats with anxiety caused by spleen-yin deficiency. It is suggested that the mechanism of action of Poria for resisting anxiety may be related to the regulation of enzyme activities of GAD, GABA-T and GS in hippocampal tissues of rats with anxiety due to spleen-yin deficiency, thereby regulating the dynamic balance of GLU and GABA content. See tables 7-8.
TABLE 7 influence of Poria decoction on GABA and GLU content in hippocampal tissue of rat anxiety model due to deficiency of spleen-yin
Group of
Dosage (g.kg)-1）
GABA(μmol·L-1)
GLU（mg·L-1）
Blank control group
—
42.66±3.779
205.24±16.66
Anxiety model group
—
38.66±4.34△
224.52±21.18
Spleen yin deficiency anxiety model group
—
34.21±3.03#
203.54±10.97
Positive control group
2.0×10-3
42.08±3.26**
191.10±23.09*
Poria cocos water decoction low dose group
4.0
42.56±4.14**
199.85±46.85
Medium dosage of Poria cocos decoction
8.0
40.07±3.07*
180.13±10.08**
Poria cocos water decoction high dose group
12.0
39.72±3.29*
198.09±31.52
Note: comparison with blank control:△P＜0.05，△△p < 0.01, compared to the anxiety model group:#P＜0.05，##p is less than 0.01; compared with the spleen yin deficiency anxiety model group: p < 0.05, P < 0.01
TABLE 8 influence of Poria decoction on GABA-T, GAD, and GS content in hippocampal tissue of rat anxiety model due to spleen-yin deficiency
Group of
Dosage (g.kg)-1）
GAD（U·L-1）
GABA-T（U·L-1）
GS（U·L-1）
Blank control group
—
238.37±29.83
192.00±9.92
472.48±16.81
Anxiety model group
—
204.77±25.18△△
208.76±10.94
419.71±26.94△△
Spleen yin deficiency anxiety model group
—
196.40±9.85
206.45±26.93
405.04±21.07
Positive control group
2.0×10-3
227.80±34.28**
180.37±16.16**
421.13±46.10
Poria cocos water decoction low dose group
4.0
220.48±18.02*
186.89±15.75*
416.74±31.19
Medium dosage of Poria cocos decoction
8.0
201.12±28.14
184.14±10.86**
443.33±37.04**
Poria cocos water decoction high dose group
12.0
200.55±8.50
190.42±5.90
416.16±22.35
Note: comparison with blank control:△P＜0.05，△△p < 0.01, compared to the anxiety model group:#P＜0.05，##p is less than 0.01; compared with the spleen yin deficiency anxiety model group: p < 0.05, P < 0.01
4. Small knot
The tuckahoe water decoction has good antianxiety effect on both spleen yang deficiency anxiety rats and spleen yin deficiency anxiety rats, and shows that the tuckahoe water decoction can be used for spleen deficiency type anxiety disorder and has clinical application value. The action mechanism is probably related to enhancing the activities of GABA synthetase and GLU degrading enzyme and inhibiting the activity of GABA degrading enzyme, thereby influencing the relative balance of GABA and GLU content, being used for preparing the anxiolytic, exploiting the medicinal value and the commercial value of tuckahoe, being an innovation on the medicine for treating anxiety disorder and having obvious economic and social benefits.
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