CN102532341B - Extraction method and application of chroogomphus rutilus polysaccharide - Google Patents
Extraction method and application of chroogomphus rutilus polysaccharide Download PDFInfo
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Abstract
The invention discloses an extraction method and application of chroogomphus rutilus polysaccharide with anti-motion sickness effects. The polysaccharide takes chroogomphus rutilus as a raw material, and is prepared by a water extraction and alcohol precipitation method. The chroogomphus rutilus polysaccharide is used for prevention and treatment of motion sickness, and the results show that very good anti-motion sickness effects can be provided when 200mg/kg of chroogomphus rutilus polysaccharide is given to the stomach by behavioral experiments and hormone level tests. The motion sickness symptoms such as visceral discomfort, incontinence, piloerection and tremor can be obviously reduced, and the level of 5-HT in serum can be reduced, and the motilin level is improved; and most importantly, rats are in good spirits, has no low spirits and other side effect symptoms. The 200mg/kg of chroogomphus rutilus polysaccharide is an ideal dose for the polysaccharide anti-motion sickness. The extraction method provides a novel material and a reference dose for further development of novel anti-motion sickness drugs with efficient, non-toxic and non-side effects.
Description
Technical field
The present invention relates to the polysaccharide of extraction from look nail mushroom [Chroogomphus rutilus] and the application in anti-kinetosis thereof, belong to health care of food and drug development technical field.
Background technology
Motion sickness is the general name of carsick, seasick, airsick and space sickness, a series of nerve of plant reactions such as show as giddy, dizzy, nauseating, vomiting, pale complexion on symptom, be in a cold sweat.It is different that its inducement is mainly that abnormal motion and low frequency noise import with the posterior infromation that is stored in central nervous system vestibular, the sense of hearing and vision into, the symptom that produces autonomic nervous dysfunction and do not feel like oneself.Although this illness is for population just sensation and spiritual discomfort, after recovering, healthy, the mental status and later work be there is no much impacts, for ride, soldier, pilot and the spacefarer of ship operation have serious harm.Because above-noted persons must keep a cool head and emergency capability in taking corresponding transportation means, otherwise the sorrow of life will be arranged and cause huge loss.The medicine of current prevention and treatment motion sickness mainly contains the moving agent of anticholinergic drug, antihistaminic, sympatheticomimetic, calcium antagonist, stomach etc., and wherein Scopolamine is that to prevent and treat at present the motion sickness effect best.Most in these medicines have the side effect that is difficult to eliminate, and as anticholinergic drug, antihistaminic, the inhibition central nervous system activities arranged, and the untoward reactions such as dry, drowsiness, blurred vision, Impairment of Memory occur; Sympatheticomimetic has the habituation side effect; The ginger herb mixture can make the stomach motion overrun; Calcium ion antagonist strongly inhibited vestibular function etc.So these medicines all can not satisfy the needs of spacefarer, pilot and soldier's anti-kinetosis.
Fungus polysaccharide is a class living polymer compound, has the multiple efficacies such as enhancing body immunizing power, anti-oxidant, antitumor, antiviral, hypoglycemic, reducing blood-fat.Research is in recent years found, pours into matsutake in Mouse Stomach, can obviously promote the emptying of Mouse Stomach; Also confirm clinically the symptoms such as that oral honey mushroom sheet can effectively be treated is dizzy, nauseating, vomiting.Infer that this inhibition is dizzy, nauseating, effect vomiting may be relevant with fungus polysaccharide.
Look nail mushroom (Chroogomphus rutilus) claims again Gomphidius rutilus, is commonly called as red mushroom, meat mushroom, pine tree umbrella, loose mushroom etc., belongs to Basidiomycotina Hymenomycetes, Agaricales, rivet mushroom section on classification of fungi.It is the important Applying Ectomycorrhizal Fungi of conifer trees.Look nail mushroom is Chinese to the maximum edible fungus of Japan outlet, is also the main raw material of cooking mushroom clear soup.Research finds, mushroom clear soup can prevent the generation of SMS effectively, but in its action component and mechanism, mushroom clear soup, the active substance of resisted motion disease be not seen in report.
Summary of the invention
The extracting method that the purpose of this invention is to provide a kind of look nail mushroom polysaccharide.
The present invention also aims to provide the application of a kind of look nail mushroom polysaccharide in anti-kinetosis.
The object of the present invention is achieved like this: the application thinks that the essential substance of resisted motion disease may be fungus polysaccharide in mushroom clear soup because maximum in the water-phase extract of mushroom be polysaccharide.More deep research has been carried out in the anti-kinetosis effect of nail mushroom polysaccharide so the applicant take rat as laboratory animal, checks colors, and result shows that this polysaccharide has remarkable anti-kinetosis effect.
Concrete, content of the present invention comprises: test experience and the application thereof of one, the extraction two of look nail mushroom polysaccharide, look nail mushroom polysaccharide anti-kinetosis effect.
The extracting method of the said look nail of the present invention mushroom polysaccharide comprises the following steps:
(1) hot water lixiviate
Clean the look nail mushroom fungus sporophore that collects with distilled water, wash three times, put into baking oven, 45 ℃ of oven dry are pulverized sporophore, weigh, and carry out heating in water bath with the water addition ratio example of water: sporophore=10ml: 1g in 80 ℃ of water-baths, and stir, extraction time 10h, centrifugal, get the supernatant vat liquor, reclaim the sporophore precipitation that obtains after centrifugal, repeat above-mentioned steps 2 times, the vat liquor merging with 3 gained is concentrated into 1/3 of gained vat liquor cumulative volume again;
(2) ethanol precipitation
It is 95% ethanol that vat liquor after concentrated is added the mass concentration of 3 times of volumes of vat liquor, and more than 4 ℃ of precipitation 10h, throw out then adds a little deionized water dissolving with absolute ethanol washing for several times, then vacuum lyophilization, i.e. Dry Sack nail mushroom polysaccharide.
As calculated, the yield of extracting institute's Dry Sack nail mushroom polysaccharide with this kind method is 6.73%.
The application of the said look nail mushroom polysaccharide of the present invention in anti-kinetosis completed by following experiment.
Below the experiment of look nail mushroom polysaccharide anti-kinetosis effect:
Take the SD rat as laboratory animal, copy the rat motor disease model with departing from the vertical axis revolving stimulation, give the rat different concns this look nail mushroom polysaccharide soln by the gavage mode before rotatory stimulation, by study of behaviour observe, asccharin water is detested experiment, spacious experiment detects, and the detection of 5-HT and motilin content in serum, estimate the effect whether this polysaccharide has the motion sickness of preventing and treating.
Copy successful motion sickness rat model, it is movable few to show as in behavior, and scope of activity is little, is slow in action, the walking phenomenon of turn-taking, also show tremble, perpendicular hair, gatism, too much modification activity etc.Carry out rotatory stimulation after giving asccharin water, make rat produce the visceral malaise symptom, to set up the detest trained reflex to asccharin water, its asccharin water of drinking is significantly reduced.Noticeable change can occur in the content of the various bioactivators in motion sickness rat model serum, and wherein judge index commonly used is 5-HT and motilin, and the 5-HT level can significantly raise, and the level of motilin can significantly descend.Give respectively to organize the look nail mushroom polysaccharide of rat various dose before rotatory stimulation, whether the mensuration by These parameters detects look nail mushroom polysaccharide the anti-kinetosis effect.
The study of behaviour that asccharin water is detested experiment, spacious experiment detects
1. animal grouping: male SD rat, body weight 200-250g is purchased from Hebei Medical University's Experimental Animal Center.Animal is bought and is divided at random 5 groups afterwards, adapts to according to experimental procedure and raises and experiment.
The grouping of table 1 animal
2. copying of motion sickness animal model: rat is placed in special fixer, is fixed on and departs from the vertical axis revolving device, after giving physiological saline or look nail mushroom polysaccharide 0.5h, with 1.5Hz rotation 1.5h.This is the international method that copies motion sickness and parameter.
3. asccharin water is detested experiment
The blank group: purchase mouse, adaptability was raised four days, freely drank water and ingested, the 5th day taboo water 24h.The 6th day morning, 8:00 freely drank asccharin water 45min, recorded the asccharin water intake.3:00 moisturizing in afternoon 45min.It is noiseless, as follows that asccharin water places rear attention mouse room peace and quiet.Blank group rat does not give rotatory stimulation, but it will be placed in by gyroscope when the physiological saline group is rotated.Observe the rat somatic reaction.
The experiment contrast group: adaptability is raised the same.Prohibited water 24h on the 5th day.The front weighing rat body weight of 8:00 in the 6th day morning and record.8:00 begins rat and freely drinks asccharin water 45min, records intake.Then according to giving physiological saline in the rat body weight stomach, physiological saline gavage amount is calculated according to 10ml/kg.After the 7th day gavage 0.5h, give rotatory stimulation 1.5h (quiet lucifuge), rotation frequency 1.5HZ.Rotation is placed on rat spacious accessible place after finishing immediately, observes the rat somatic reaction, returns cage.The 8th day morning, 8:00 measured asccharin water intake and record in rat 45min, surveyed continuously 5 days.
The fungus polysaccharide experimental group: adaptability is raised the same.Prohibited water 24h on the 5th day.The front weighing rat body weight of 8:00 in the 6th day morning and record, 8:00 begins rat and freely drinks asccharin water 45min, records intake.Then according to rat body weight gavage respective concentration fungus polysaccharide, the concentration setting is respectively 100mg/kg, 200mg/kg, 400mg/kg, and the gavage amount converts according to rat body weight.Give rotatory stimulation after the 7th day gavage 0.5h.Rotation is placed on rat spacious accessible place after finishing immediately, observes the rat somatic reaction, returns cage.The 8th day morning, 8:00 measured asccharin water intake and record in rat 45min, surveyed continuously 5 days.
Data statistics: observe somatic reaction in rat 10min after rotation finishes; Relatively rat is rotated the day before yesterday and rear three days asccharin water intake difference of rotation.Experimental result represents with mean number ± standard deviation, carries out the t check with the STATISTICA6.0 Software of Data Statistics, and P＜0.05 shows that result has statistical significance.
4. spacious experiment
Blank group: adapt to when raising and freely drink water, ingest, rotate the concluding time according to rotation group rat rat is put into spacious 15min successively according to numbering, make it be familiar with spacious and surrounding enviroment, for three days on end.Control rats does not give rotatory stimulation (being placed in by instrument) when the experiment contrast group gives rotatory stimulation, then rat is put into spacious center lattice, makes a video recording simultaneously and timing, finishes after 10min.Record the residence time (latent period), the modification time of rat in center lattice, as follows.Keep the surrounding environment peace and quiet noiseless during shooting.(annotate: no matter be to adapt to raise or formal experiment, after rat is taken out from spacious, ight soil in certain in time wash-up tray is with wet towel or at the bottom of dipping in spirituous wet cloth wiping case and wall, in order to avoid the stool and urine of a upper rat or the ensuing test result of smell impact.As follows)
Experiment contrast group: adapt to when raising and freely drink water, ingest, rotate the concluding time according to the rat that calculates rat is put into spacious 15min successively according to numbering, make it be familiar with spacious and surrounding enviroment, for three days on end.Give physiological saline in the 4th day rat stomach, the gavage amount is calculated according to 100mg/kg, continues according to the set time, rat to be put into spacious and adapts to.After the 5th day gavage 0.5h, give rotatory stimulation.After rotation finishes, rat is put into spacious center lattice, make a video recording simultaneously and timing, finish after 10min.Record correlation parameter.
Fungus polysaccharide experimental group: adapt to when raising identical with the experiment contrast group.The concentration setting was respectively 100mg/kg, 200mg/kg, 400mg/kg according to rat body weight gavage respective concentration fungus polysaccharide in the 4th day, and the gavage amount converts according to rat body weight, continued according to the set time, rat to be put into spacious and adapted to.After the 5th day gavage 0.5h, give rotatory stimulation.After rotation finishes, rat is put into spacious center lattice, make a video recording simultaneously and timing, finish after 10min.Record correlation parameter.
Data statistics: in experiment, 9 lattice of spacious bottom surface central authorities are regarded as center lattice, rat in residence time of center lattice as latent period.Record the rat time in latent period.Rat licks hair, licks foot, two forelimbs reason hairs, comfort the modification that the action such as mouth is called rat, the modification time of the interior rat of record shooting 10min.Experimental result represents with mean number ± standard deviation, carries out the t check with the STATISTICA6.0 Software of Data Statistics, and P＜0.05 shows that result has statistical significance.
The detection of 5-HT and motilin content in serum
1. animal grouping: male SD rat, body weight are between 200--250g, and medical university's experimental animal center, effluent north is bought.Adapt to raising and be divided at random afterwards 5 groups in 3 days, the situation of specifically dividing into groups is as shown in table 2.
The grouping of table 2 animal and processing
2. the copying and taking a blood sample of motion sickness animal model: above-mentioned different treated animals, after giving physiological saline or look nail mushroom polysaccharide 0.5h, rat is placed in special fixer, be fixed on and depart from the vertical axis revolving device, with 1.5Hz rotation 1.5h.After rotation finishes immediately anesthetized rat (vetanarcol (60mg/kg) are cut off the thoracic cavity, expose heart, heart blood sampling 6ml, and rapidly with blood transfer in the 5ml centrifuge tube, a rat blood bifurcation centrifuge tube is deposited.After hemopexis, get upper serum to another centrifuge tube with micro sample adding appliance, mark is placed in-20 ℃ of refrigerators frozen to be measured.
3. ELISA kit measurement: the 5-HT and the motilin test kit that are provided by Shanghai Hu Feng bio tech ltd, and entrust the said firm to detect the level of respectively organizing two kinds of active substances in serum with double-blind method.
4. data statistics: experimental result represents with mean number ± standard deviation, carries out the t check with the STATISTICA6.0 Software of Data Statistics, and P＜0.05 shows that result has statistical significance.
The study of behaviour detected result of look nail mushroom polysaccharide anti-kinetosis effect
Outward appearance behavior observation result
The blank group: this treated animal does not award intragastric infusion physiological saline, does not carry out the motion sickness model copy yet, only is placed on to depart from vertical axis revolving stimulating instrument side.Show as movable sensitivity, exploratory behaviour is many always, and feed, drinking-water are normally.
Experiment contrast group: after giving the physiological saline processing in this treated animal stomach, carry out the motion sickness model copy.Behavior before copying is identical with the blank group.After departing from vertical axis revolving (1.5Hz, 1.5h), show significant motion sickness symptom, as few in activity, scope of activity is little, and behavior is blunt, and the walking phenomenon of turn-taking occurs trembling, perpendicular hair, gatism, too much grooming behavior etc.Show that copying of motion sickness model is successfully.
Experimental group: give respectively 100mg/kg, 200mg/kg, 400mg/kg look nail mushroom polysaccharide in this treated animal stomach.Copy the motion sickness model front identical with the behavior of blank treated animal.After giving rotatory stimulation, the animal that gives 200mg/kg look nail mushroom polysaccharide in stomach without gatism, tremble and erect a mao phenomenon, and energetic, without dispirited symptom, grooming behavior, Exploratory behavior are also identical with the blank treated animal; After giving rotatory stimulation, the animal that gives respectively 100mg/kg, 400mg/kg look nail mushroom polysaccharide in stomach shows slight motion sickness behavior, as moving and the phenomenons such as exploratory behaviour is less, perpendicular hair.This shows, giving 200mg/kg look nail mushroom polysaccharide in stomach has good anti-kinetosis effect, the most important thing is that the rat mental status is good, is a cup too low dispirited and other side effect symptom.
Asccharin water is detested the detected result of experiment, spacious experiment
1. asccharin water is detested laboratory test results
As shown in table 3, Fig. 1-5.The stable significance that do not occur of the asccharin water intake of blank group rat changes; Considerable change occurs in experiment contrast group rat asccharin water intake before and after rotation: compare the day before yesterday with rotation, rotate the asccharin water intake of rear first day and second day and turn front level error heteropole significantly (P＜0.01), although after rotation, the 3rd day asccharin water intake increases to some extent, with turn before compare and significant difference still arranged (P＜0.05); Give look nail mushroom polysaccharide soln in stomach, the asccharin water intake of 100mg/kg polysaccharide group rat only after rotation first day with turn before there were significant differences (P＜0.05), second day can return to level before rotation.200mg/kg polysaccharide group rat does not show detest to asccharin water afterwards in rotation, the asccharin water intake before and after rotation without significant difference (P＞0.05).After 400mg/kg polysaccharide group rat rotation the asccharin water intake of first day with turn before compare difference extremely significantly (P＜0,01), after rotation, second day begins the asccharin water intake and significantly improves, and returns to gradually level before rotation.
The impact of table 3X fungus polysaccharide solution on rat asccharin water intake
Annotate: in group relatively, a representative with turn the day before yesterday of P＜0.01 relatively, the b representative with turn the day before yesterday of P＜0.05 relatively, the c representative with turn rear first day relatively P＜0.05, the d representative with turn rear first day relatively P＜0.01.The mark of following Fig. 1-Fig. 5 is identical therewith.
Annotate: in group relatively, a representative with turn the day before yesterday of P＜0.01 relatively, the b representative with turn the day before yesterday of P＜0.05 relatively, the c representative with turn rear first day relatively P＜0.05, the d representative with turn rear first day relatively P＜0.01.
This experimental result demonstration, the look nail mushroom polysaccharide that gives 200mg/kg before rotatory stimulation can alleviate rat to the detest of asccharin water, shows that this polysaccharide can improve the visceral malaise symptom.
2. spacious laboratory test results
After this experiment has detected rotatory stimulation and has finished, the latent period of animal from the outage to the activity, the modification time in 10min.Result shows (table 4, shown in Fig. 6-7), significantly be longer than blank group (P＜0.05) latent period of experimental group rat, the latent period of 100mg/kg, 200mg/kg polysaccharide group rat and blank group are without significant difference (P＞0.05), and the utmost point in latent period of 400mg/kg polysaccharide group rat is longer than blank group (P＜0.01) significantly; On the modification time, the experiment contrast group significantly is longer than blank group (P＜0.05), 100mg/kg, 200mg/kg polysaccharide group and blank group are without significant difference (P＞0.05), and 400mg/kg polysaccharide group significantly is longer than blank group (P＜0.05).
Spacious experimental result of table 4 rat
Annotate: a and represent with the blank group and compare P＜0.05, the b representative is compared P＜0.01 with the blank group, and c representative and 400mg/kg polysaccharide group be P＜0.01 relatively, and d representative and experiment contrast group be P＜0.05 relatively.The mark of following Fig. 6-Fig. 7 is identical therewith.
This detected result shows, gives 100mg/kg before rotatory stimulation, 200mg/kg look nail mushroom polysaccharide can significantly alleviate the motion sickness symptom of rat, make grooming behavior that its motor capacity and visceral malaise show all with the rat that there is no motion sickness without significant difference.
The detected result of 5-HT and motilin content in serum
The detected result of 5-HT shows (table 5 and Fig. 8,9), and in the serum of experiment contrast group rat, 5-HT content significantly improves (P＜0.05) than blank group rat; Give in the serum of look nail mushroom polysaccharide 5-HT content all low than experiment contrast group rat, and compare without significant difference (P＞0.05) with blank group rat.In 200mg/kg polysaccharide group rat blood serum, 5-HT content significantly reduces (P＜0.05) than experiment contrast group rat; In 100mg/kg and 400mg/kg polysaccharide group rat blood serum, 5-HT content and experiment contrast group are without marked difference (P＜0.05).
The detected result of motilin shows (table 5 and Fig. 8,9), and experiment contrast group rat stomach therbligs content significantly reduces (P＜0.05) than blank group rat; The motilin content of 100mg/kg, 200mg/kg, 400mg/kg polysaccharide group rat is compared there was no significant difference (P＞0.05) with the blank group; Compare with the experiment contrast group, 100mg/kg polysaccharide group rat stomach therbligs content significantly improves (P＜0.05), and 200mg/kg, the 400mg/kg polysaccharide group rat stomach therbligs content utmost point significantly improve (P＜0.01).
The impact of table 5 look nail mushroom polysaccharide on motion sickness rat blood serum 5-HT and motilin content
Annotate: a representative is compared P＜0.05 with the blank group; The b representative is compared P＜0.05 with the experiment contrast group; The c representative is compared P＜0.01 with the experiment contrast group; The d representative is compared P＜0.05 with 200mg/kg polysaccharide group.The mark of Fig. 8-Fig. 9 is identical therewith.
This detected result shows, gives rising that 100mg/kg, 200mg/kg look nail mushroom polysaccharide can significantly suppress the serum 5-HT content that caused by rotatory stimulation and the reduction of motilin content before rotatory stimulation.
Above-mentioned all detected results all show, giving 200mg/kg look nail mushroom polysaccharide in stomach has good anti-kinetosis effect.Not only can significantly alleviate the motion sickness symptom, as visceral malaise, gatism, perpendicular hair, tremble etc., can also reduce 5-HT level in serum, improve motilin levels, the most important thing is that the rat mental status is good, be a cup too low dispirited and other side effect symptom.Show that 200mg/kg look nail mushroom polysaccharide is the comparatively desirable dosage of this polysaccharide anti-kinetosis.So the present invention provides new material and reference dose for further developing anti-kinetosis new drug efficient, that have no side effect.
Above-mentioned experiment confirm look nail mushroom polysaccharide can be used for preparing motion sickness resistant medicament.
Description of drawings
Fig. 1 is blank group asccharin water intake.
Fig. 2 is experiment contrast group asccharin water intake.
Fig. 3 is that 100mg/kg polysaccharide group asccharin water is taken in.
Fig. 4 is 200mg/kg polysaccharide group asccharin water intake.
Fig. 5 is 400mg/kg polysaccharide group asccharin water intake.
Fig. 6 respectively organizes the rat rotation to compare rear latent period.
Fig. 7 respectively organizes the rear modification time of rat rotation relatively.
Fig. 8 respectively organizes 5-HT level comparison in rat blood serum.
Fig. 9 is the comparison of respectively organizing motilin content in rat blood serum.
Embodiment
Following examples are used for explanation the present invention.
Embodiment
One, the extraction of look nail mushroom polysaccharide
(1) hot water lixiviate
Get the look nail mushroom fungus sporophore that 680g collects, with distilled water wash three times, put into baking oven, 45 ℃ of oven dry, dry Dry Sack nail mushroom fungus sporophore 650g is again with its pulverizing, water addition ratio example with water: sporophore=10ml: 1g is carried out heating in water bath in 80 ℃ of water-baths, and stir, extraction time is 10h, and is centrifugal, gets the supernatant vat liquor, reclaim the sporophore precipitation that obtains after centrifugal, repeat again above-mentioned steps 2 times, the vat liquor of 3 gained merged, and be concentrated in Rotary Evaporators 3 gained the vat liquor cumulative volume 1/3;
(2) ethanol precipitation
It is 95% ethanol that vat liquor after concentrated is added the mass concentration of 3 times of volumes, and more than 4 ℃ of precipitation 10h, throw out then adds a little deionized water dissolving with absolute ethanol washing for several times, then vacuum lyophilization Dry Sack nail mushroom polysaccharide 43.64g.
Two, look nail mushroom polysaccharide anti-kinetosis application experiment
Take spacious experiment-latent period as example.
Purchase 30 of SD rats from Hebei Medical University, body weight is between 200--250g.Adapt to raising and be divided at random afterwards 5 groups in 3 days, the situation of specifically dividing into groups is as shown in table 6.
Annotate: k represents spacious experiment; K represents the blank group; S represents the experiment contrast group; 100 represent 100mg/kg polysaccharide group; 200 represent 200mg/kg polysaccharide group; 400 represent 400mg/kg polysaccharide group.
The blank group: adapt to when raising and freely drink water, ingest, every morning 10:18 puts into spacious 15min, makes it be familiar with spacious and surrounding enviroment, for three days on end.The 5th day morning 8:48, blank group rat is placed in when the experiment contrast group gives rotatory stimulation by instrument, 10:18 puts into spacious center lattice with blank group rat, makes a video recording simultaneously and timing, finishes after 10min.Record the residence time (latent period time) of rat in center lattice.Keep the surrounding environment peace and quiet noiseless during shooting.(annotate: no matter be to adapt to raise or formal experiment, after rat is taken out from spacious, ight soil in certain in time wash-up tray is with wet towel or at the bottom of dipping in spirituous wet cloth wiping case and wall, in order to avoid the stool and urine of a upper rat or the ensuing test result of smell impact.As follows)
The experiment contrast group: adapt to when raising and freely drink water, ingest, every morning 10:00 puts into spacious 15min, makes it be familiar with spacious and surrounding enviroment, for three days on end.Early gave physiological saline in the 8:00 rat stomach on the 4th day, the gavage amount is calculated according to 100mg/kg, continues according to the set time, rat to be put into spacious and adapts to.The 5th day morning, the 8:00 gavage, after 0.5h, gave rotatory stimulation 1.5h.The morning, 10:00 put into spacious center lattice with rat, made a video recording simultaneously and timing, finished after 10min.Record correlation parameter.
Fungus polysaccharide experimental group: adapt to when raising and freely drink water, ingest, 100mg/kg polysaccharide group 11:30 at noon every day puts into spacious 15min, 200mg/kg polysaccharide group 13:00 every afternoon puts into spacious 15min, 400mg/kg polysaccharide group 14:30 every afternoon puts into spacious 15min, make it be familiar with spacious and surrounding enviroment, for three days on end.Adapt to when raising identical with the experiment contrast group.The concentration setting was respectively 100mg/kg, 200mg/kg, 400mg/kg according to rat body weight gavage respective concentration fungus polysaccharide in the 4th day, and the gavage amount converts according to rat body weight, continued according to the set time, rat to be put into spacious and adapted to.After the 5th day gavage 0.5h, give rotatory stimulation 1.5h.After rotation finishes, rat is put into spacious center lattice, make a video recording simultaneously and timing, finish after 10min.Record correlation parameter.
Data statistics: in experiment, 9 lattice of spacious bottom surface central authorities are regarded as center lattice, rat in residence time of center lattice as latent period.Record the rat time in latent period.Concrete experimental result sees Table 7.
Annotate: k represents spacious experiment; K represents the blank group; S represents the experiment contrast group; 100 represent 100mg/kg polysaccharide group; 200 represent 200mg/kg polysaccharide group; 400 represent 400mg/kg polysaccharide group.Experimental result represents with mean number ± standard deviation, carries out the t check with the STATISTICA6.0 Software of Data Statistics, and P＜0.05 shows that result has statistical significance.Each is organized the rat rotation and relatively sees afterwards Fig. 6 latent period.
Claims (2)
1. the extracting method of look nail mushroom polysaccharide is characterized in that extracting method carries out according to the following steps:
(1) hot water lixiviate
Clean the look nail mushroom fungus sporophore that collects with distilled water, wash three times, put into baking oven, 45 ℃ of oven dry are pulverized sporophore, weigh, and with water: the water addition ratio example of sporophore=10 ml:1 g is carried out heating in water bath in 80 ℃ of water-baths, and stir, extraction time is 10 h, and is centrifugal, gets the supernatant vat liquor, reclaim the sporophore precipitation that obtains after centrifugal, repeat above-mentioned steps 2 times, the vat liquor merging with 3 gained is concentrated into 1/3 of gained vat liquor cumulative volume again;
(2) ethanol precipitation
It is 95% ethanol that vat liquor after concentrated is added the mass concentration of 3 times of volumes, and more than 4 ℃ of precipitation 10h, throw out then adds a little deionized water dissolving with absolute ethanol washing for several times, and then, vacuum lyophilization is Dry Sack nail mushroom polysaccharide.
2. the application of a look nail mushroom polysaccharide, the look nail mushroom polysaccharide that it is characterized in that extracting by claim 1 is for the preparation of motion sickness resistant medicament.
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| CN102018731B (en) * | 2010-11-17 | 2013-04-03 | 河北师范大学 | Method for extracting anti-oxidation active substances from chroogomphus rutilus and application of obtained extracts |
| CN102002113A (en) * | 2010-12-08 | 2011-04-06 | 江南大学 | Method for preparing tea plant mushroom fruit body polysaccharide |
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