CN103751213B - A kind of earthworm extract and application thereof - Google Patents
A kind of earthworm extract and application thereof Download PDFInfo
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- CN103751213B CN103751213B CN201310565898.2A CN201310565898A CN103751213B CN 103751213 B CN103751213 B CN 103751213B CN 201310565898 A CN201310565898 A CN 201310565898A CN 103751213 B CN103751213 B CN 103751213B
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Abstract
The invention discloses a kind of earthworm extract and application thereof.Described earthworm extract be molecular weight be the water extract of the Lumbricus of below 3000 dalton, described Lumbricus is one or more in William's cavity ring earthworm, Eisenia foetida and Amythas dancatala.The earthworm extract of the present invention has the function significantly reduced blood pressure, and its preparation method is simple to operate, it is possible to achieve prepare on a large scale.
Description
The application is filing date on October 12nd, 2010, Application No. 201010503564.9, invention entitled " a kind of
Earthworm extract, its preparation method, apply and containing its pharmaceutical composition " the divisional application of patent application.
Technical field
The present invention is specifically related to a kind of earthworm extract and application thereof.
Background technology
Lumbricus is commonly called as Pheretima, have antipyretic, relieving convulsion, blood pressure lowering, antibacterial, press down the effect such as cancer and blood circulation promoting and blood stasis dispelling, the most extensively apply
Treating multiple disease in the traditional Chinese medical science, especially during treatment hypertension, Pheretima has become in Chinese medicine simply the most
Indispensable medical material.Compel to be essential and it is carried out basis and exploitation applied research.
Summary of the invention
The technical problem to be solved there is provided to be had a kind of earthworm extract of hypotensive activity and answers
With.The earthworm extract of the present invention has the function significantly reduced blood pressure, and its preparation method is simple to operate, it is possible to achieve big
The preparation of scale.
The present inventor finds through numerous studies, and the material that can play hypotensive activity in the Chinese medicine decoction of Lumbricus is little
The material of the albumen in the earthworm extract of molecular weight rather than Lumbricus etc.But, at the Lumbricus compound of these little molecules
Middle can play hypotensive activity active ingredient be actually the mechanism of action of what and it how, be a class urgently to be resolved hurrily
Topic.The present inventor, through research, is first found that small-molecular-weight earthworm extract that a class is new and a set of extensive preparation should
The method of earthworm extract.
Therefore the present invention relates to a kind of earthworm extract, it is the water extract of Lumbricus, molecular weight be 3000 dalton with
Under.
The invention further relates to the preparation method of above-mentioned earthworm extract, it comprises the steps of takes out the water of Lumbricus
Extract, through 3000 molecular weight ultrafiltration through membranes, collects filtrate,.
Wherein, described Lumbricus can be can be used as medicine in this area use various Lumbricuss, be preferably William's cavity ring earthworm,
One or more in Eisenia foetida and Amythas dancatala.
In the preparation method of the present invention, the water extract of described Lumbricus can be by conventional the having in Lumbricus with water in this area
The method that effect composition carries out extracting prepares, it is also preferred that the left it is prepared by following method: smash by the Lumbricus after telling mud, washing process
Be broken into pulpous state, soak after the mixing that adds water, then with the preferred 1000-2000 of 1000-3000(, more preferably 1500) rev/min centrifugal
Machine is centrifuged, and obtains supernatant 1, by the supernatant 1 preferred 15000-17000 of 10000-20000(, more preferably 16000) rev/min from
Scheming is centrifuged, and obtains supernatant 2,.
Wherein, the described amount added water is preferably for 0.8-1.5 times (preferably 1 times) of pulpous state Lumbricus volume；Soak time
Between be preferably 3-10 hour (preferably 3-5 hour, more preferably 3 hours)；The temperature soaked preferably is 4-10 DEG C；On obtaining
After clear liquid 2, the most also comprise drying steps, such as vacuum lyophilization.
In the preparation method of the present invention, it is also preferred that the left also comprised the following steps: before with 3000 molecular weight ultrafiltration through membranes by
After the water extract of Lumbricus is dissolved in water, then with centrifuge with the preferred 15000-17000 of 10000-20000(, more preferably 16000)
Rev/min centrifugal, obtain supernatant；The water extract of described Lumbricus is dried object.Wherein, the described amount added water is preferably for earthworm
1-5 times (preferably 2 times) of the water extract volume of earthworm.Described dried object is the dried material of water extract of Lumbricus.
With after 3000 molecular weight ultrafiltration through membranes, also can comprise the steps of and gained material is dried,.Described
It is dried preferably for vacuum lyophilization.
The invention further relates to a kind of drug regimen comprising above-mentioned earthworm extract and pharmaceutically acceptable carrier
The pharmaceutical composition of acceptable carrier or health food on thing or bromatology.
In the pharmaceutical composition of the present invention, above-mentioned earthworm extract can play a role as single active component, also
Can according to common sense in the field with possess the functions such as blood pressure lowering, and pharmaceutically active with earthworm extract generation untoward reaction does not becomes
Divide and play a role together as active component.
The earthworm extract of the present invention can make medicine with the most various typical additives (such as diluent and excipient etc.)
Compositions.According to therapeutic purposes, pharmaceutical composition can be made various types of administration unit dosage forms, such as tablet, pill, powder
Agent, liquid, suspension, emulsion, granule, capsule, suppository and injection (solution and suspension) etc..
In order to make the pharmaceutical composition of tablet form shape, any of and widely used figuration in this area can be used
Agent.Such as, carrier, such as lactose, white sugar, sodium chloride, glucose, carbamide, starch, calcium carbonate, Kaolin, crystalline cellulose and silicon
Acid etc.；Binding agent, such as water, ethanol, propanol, common syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose
Element, lac, methylcellulose and potassium phosphate, polyvinylpyrrolidone etc.；Disintegrating agent, such as dried starch, sodium alginate, agar powder and sea
Band powder, sodium bicarbonate, calcium carbonate, the fatty acid ester of polyethylene sorbitan, sodium lauryl sulphate, stearic acid monoglycerides,
Starch and lactose etc.；Disintegrate inhibitor, such as white sugar, glycerol tristearate, Oleum Cocois and hydrogenated oil and fat；Adsorption enhancer, such as season
Amine base and sodium lauryl sulphate etc.；Wetting agent, such as glycerol, starch etc.；Adsorbent, such as starch, lactose, Kaolin, bentonite
With colloid silicic acid etc.；And lubricant, such as pure Talcum, stearate, boric acid powder and Polyethylene Glycol etc..If needed
Words, it is also possible to make tablet as sugar coated tablet with common coated material, be coated with gelatin film tablet, enteric coated tablets, film coated tablets, double
Tunic tablet and multilayer tablet.
In order to make the pharmaceutical composition of pill shape, any of and widely used inborn nature in this area can be used
Agent, such as, carrier, such as lactose, starch, Oleum Cocois, hardened vegetable oils, Kaolin and Talcum etc.；Binding agent, such as Radix Acaciae senegalis
Powder, yellow work rubber powder, gelatin and ethanol etc.；Disintegrating agent, such as agar and Kelp Powder etc..
In order to make the pharmaceutical composition of suppository form shape, this area can be used any known and widely used inborn nature
Agent, such as, Polyethylene Glycol, Oleum Cocois, higher alcohol, the ester of higher alcohol, gelatin and semisynthetic glyceride etc..
In order to prepare the pharmaceutical composition of injection form, solution and suspension can be sterilized, and be preferably added appropriate chlorine
Change sodium, glucose or glycerol etc., make injection isotonic with blood.When preparing injection, it is possible to use any in this area
Conventional carrier.Such as, water, ethanol, propylene glycol, the isooctadecanol of ethoxylation, the isooctadecanol of polyoxy and polyethylene
The fatty acid ester etc. of anhydro sorbitol.Additionally, common lytic agent, buffer agent and analgesic etc. also can be added.As required, exist
During treatment schizophrenia, it is possible to add coloring agent, preservative, spice, flavoring agent, sweetening agent and other medicines etc..
The earthworm extract of present invention content in pharmaceutical composition, without particular restriction, can be carried out in a wide range
Select.
In the present invention, the medication of described pharmaceutical composition is not particularly limited.Can according to patient age, sex and
Other condition and symptom, select the preparation of different dosage form to be administered.Such as, tablet, pill, solution, suspension, emulsion, granule
It is oral administration with capsule；Injection (being preferably aqueous injection or lyophilized injectable powder) can be individually dosed, or defeated with injection
Liquor charging (such as glucose solution and Freamine Ⅲ) is mixed into row vein injection, can carry out flesh by simple injection if necessary
Meat, Intradermal, the subcutaneous or interior injection of abdomen；Suppository is for being administered into rectum.
In the present invention, use can be properly selected according to method of administration, patient age, sex and other condition and symptom
Pharmaceutical quantities.
The invention still further relates to the application in the medicine preparing blood pressure lowering of the above-mentioned earthworm extract.
The invention still further relates to above-mentioned earthworm extract and prepare treatment or the health food of prophylaxis of hypertension or functional food
Application in product.
In addition to specified otherwise, the raw material that the present invention relates to and reagent are the most commercially.
The most progressive effect of the present invention is: the earthworm extract of the present invention has the function significantly reduced blood pressure,
And its preparation method is simple to operate, it is possible to achieve prepare on a large scale.
Detailed description of the invention
Further illustrate the present invention below by the mode of embodiment, but the most therefore limit the present invention to described reality
Execute among example scope.The experimental technique of unreceipted actual conditions in the following example, conventionally and condition, or according to business
Product description selects.
Embodiment 1: the extraction of William's cavity ring earthworm water extract
1, fresh William's cavity ring earthworm (1 kilogram) is through telling mud, and water puts into freezer frost after cleaning, standby.
2, being taken out from freezer by William's cavity ring earthworm of frost, after naturally thawing, pulp smashed to pieces by machinery.Add 1 times of volume
Water mixes, and with the centrifuge 10 minutes of 1500 revs/min after soaking 3 hours, leaves supernatant, discards precipitation.
3, supernatant is left after being centrifuged by supernatant 16000 revs/min of the machine by centrifugation of 2.
4, by 3 supernatant vacuum lyophilization, i.e. can get William's cavity ring earthworm water extract.
Embodiment 2: the extraction of William's cavity ring earthworm extract
1, the Lumbricus water extract that Example 1 prepares is centrifuged with 16000 revs/min of centrifuge after adding the water dissolution of 2 times.Stay
Lower supernatant.
2, by 1 the supernatant ultrafilter ultrafiltration of 3000 molecular weight ultrafilter membranes, available two kinds of components, respectively 3000
Water soluble materials (hereinafter referred to as " mixture A ") (29.8 grams) below molecular weight film and aqueous solutions more than 3000 molecular weight films
Material (hereinafter referred to as " mixture B ").
3, by 2 mixture A vacuum lyophilization, obtain the extract of William's cavity ring earthworm highly finished product, that is the present invention divide
Son amount William's cavity ring earthworm extract below 3000 dalton.
4, by 2 mixture B vacuum lyophilization, standby.
Embodiment 3: in example 2, uses the mixture A that the ultrafilter membrane ultra-filtration and separation of 3000 molecular weight obtains, therefore demonstrate,proves
The mixture A obtained after this ultrafiltration bright is that molecular weight is less than 3000 daltonian William's cavity ring earthworm earthworm extracts.
Embodiment 4: the extraction of Eisenia foetida water extract
1, fresh Eisenia foetida (1 kilogram) is through telling mud, and water puts into freezer frost after cleaning, standby.
2, being taken out from freezer by the Eisenia foetida of frost, after naturally thawing, pulp smashed to pieces by machinery.Add 1 times of volume
Water mixes, and with the centrifuge 10 minutes of 1500 revs/min after soaking 3 hours, leaves supernatant, discards precipitation.
3, supernatant is left after being centrifuged by supernatant 16000 revs/min of the machine by centrifugation of 2.
4, by 3 supernatant vacuum lyophilization, i.e. can get Eisenia foetida water extract.
Embodiment 5: the extraction of Eisenia foetida extract
1, after the Lumbricus water extract goods that Example 4 prepares add the water dissolution of 2 times with 16000 revs/min of centrifuge from
The heart.Leave supernatant.
2, by 1 the supernatant ultrafilter ultrafiltration of 3000 molecular weight ultrafilter membranes, available two kinds of components, respectively 3000
Water soluble materials (hereinafter referred to as " mixture C ") (28.7 grams) below molecular weight film and aqueous solutions more than 3000 molecular weight films
Material (hereinafter referred to as " mixture D ").
3, by 2 mixture C vacuum lyophilization, obtain the extract of Eisenia foetida highly finished product, that is the present invention divide
Son amount Eisenia foetida extract below 3000 dalton.
4, by 2 mixture D vacuum lyophilization, standby.
Embodiment 6: in embodiment 5, uses the mixture C that the ultrafilter membrane ultra-filtration and separation of 3000 molecular weight obtains, therefore demonstrate,proves
The mixture C obtained after this ultrafiltration bright is that molecular weight is less than 3000 daltonian Eisenia foetida extracts.
Embodiment 7: the extraction of Amythas dancatala water extract
1, fresh Amythas dancatala (1 kilogram) is through telling mud, and water puts into freezer frost after cleaning, standby.
2, being taken out from freezer by the Amythas dancatala of frost, after naturally thawing, pulp smashed to pieces by machinery.Add the water of 1 times of volume
Mixing, with the centrifuge 10 minutes of 1500 revs/min after soaking 3 hours, leaves supernatant, discards precipitation.
3, supernatant is left after being centrifuged by supernatant 16000 revs/min of the machine by centrifugation of 2.
4, by 3 supernatant vacuum lyophilization, i.e. can get Amythas dancatala goods water extract.
Embodiment 8: the extraction of Amythas dancatala extract
1, after the Amythas dancatala water extract that Example 7 prepares adds the water dissolution of 2 times with 16000 revs/min of centrifuge from
The heart.Leave supernatant.
2, by 1 the supernatant ultrafilter ultrafiltration of 3000 molecular weight ultrafilter membranes, available two kinds of components, respectively 3000
Water soluble materials (hereinafter referred to as " mixture E ") (29.1 grams) below molecular weight film and aqueous solutions more than 3000 molecular weight films
Material (hereinafter referred to as " mixture F ").
3, by 2 mixture E vacuum lyophilization, obtain the extract of Amythas dancatala highly finished product, that is molecule of the present invention
Amount Amythas dancatala extract below 3000 dalton.
4, by 2 mixture F vacuum lyophilization, standby.
Embodiment 9: in embodiment 8, uses the mixture E that the ultrafilter membrane ultra-filtration and separation of 3000 molecular weight obtains, therefore demonstrate,proves
The mixture E obtained after this ultrafiltration bright is that molecular weight is less than 3000 daltonian Amythas dancatala extracts.
Embodiment 10: the extraction of William's cavity ring earthworm water extract
1, fresh William's cavity ring earthworm (1 kilogram) is through telling mud, and water puts into freezer frost after cleaning, standby.
2, being taken out from freezer by William's cavity ring earthworm of frost, after naturally thawing, pulp smashed to pieces by machinery.Add 0.8 times of volume
Water mixing, after soaking 10 hours, (soaking temperature 4 DEG C) is with the centrifuge 10 minutes of 1000 revs/min, leaves supernatant, abandons
Fall precipitation.
3, supernatant is left after being centrifuged by supernatant 10000 revs/min of the machine by centrifugation of 2.
4, by 3 supernatant vacuum lyophilization, i.e. can get William's cavity ring earthworm water extract.
Embodiment 11: the extraction of William's cavity ring earthworm extract
1, the Lumbricus water extract that Example 10 prepares is centrifuged with 10000 revs/min of centrifuge after adding the water dissolution of 1 times.
Leave supernatant.
2, by 1 the supernatant ultrafilter ultrafiltration of 3000 molecular weight ultrafilter membranes, available two kinds of components, respectively 3000
Water soluble materials (hereinafter referred to as " mixture A1 ") (29 grams) below molecular weight film and aqueous solutions more than 3000 molecular weight films
Material (hereinafter referred to as " mixture B1 ").
3, by 2 mixture A vacuum lyophilization, obtain the extract of William's cavity ring earthworm highly finished product, that is the present invention divide
Son amount William's cavity ring earthworm extract below 3000 dalton.
4, by 2 mixture B1 vacuum lyophilization, standby.
Embodiment 12: the extraction of Eisenia foetida water extract
1, fresh Eisenia foetida (1 kilogram) is through telling mud, and water puts into freezer frost after cleaning, standby.
2, being taken out from freezer by the Eisenia foetida of frost, after naturally thawing, pulp smashed to pieces by machinery.Add 1.5 times of volumes
Water mixing, after soaking 6 hours, (soaking temperature 10 DEG C) is with the centrifuge 10 minutes of 3000 revs/min, leaves supernatant, abandons
Fall precipitation.
3, supernatant is left after being centrifuged by supernatant 20000 revs/min of the machine by centrifugation of 2.
4, by 3 supernatant vacuum lyophilization, i.e. can get Eisenia foetida water extract.
Embodiment 13: the extraction of Eisenia foetida extract
1, after the Lumbricus water extract goods that Example 12 prepares add the water dissolution of 5 times with 20000 revs/min of centrifuge from
The heart.Leave supernatant.
2, by 1 the supernatant ultrafilter ultrafiltration of 3000 molecular weight ultrafilter membranes, available two kinds of components, respectively 3000
More than water soluble materials (hereinafter referred to as " mixture C 1 ") (28.6 grams) below molecular weight film and 3000 molecular weight films water-soluble
Liquid material (hereinafter referred to as " mixture D 1 ").
3, by 2 mixture C 1 vacuum lyophilization, obtain the extract of Eisenia foetida highly finished product, that is the present invention divide
Son amount Eisenia foetida extract below 3000 dalton.
4, by 2 mixture D 1 vacuum lyophilization, standby.
Comparative example 1: the blood pressure lowering experiment of mixture A and mixture B
1 prepares sample
(1) mixture A, B redistilled water is configured to three different concentration.
Mixture A:8mg/ml 4mg/ml 2mg/ml
Mixture B:8mg/ml 4mg/ml 2mg/ml
(2) set redistilled water and make negative control
2 experimental techniques
Take rat, with 20 urethane anesthesia after weighing.Face upward position to fix, separate common carotid artery, intubate recording blood pressure (instrument
Device: RM6000 tetra-leads physiograph, AP-601, AP-611 recording blood pressure), the diastolic pressure of animal and contraction pressure are direct by instrument
Read.
Post operation stablizes 30min, recording blood pressure.Gavage is dissolved in mixture A, B of redistilled water, two kinds of mixture respectively
Dosage is respectively 40mg/Kg, 20mg/Kg, 10mg/Kg, additionally sets gavage and gives redistilled water and make negative control.Giving mixture
30min after A, B, 60min, 90min, 120min, 150min, 180min recording blood pressure respectively.
3 data process
Calculating average and the standard deviation shrinking pressure and diastolic pressure respectively, between group, difference uses t check analysis.Mixture A with
P ＜ 0.05 between mixture B, there were significant differences.
4 results (see table 1 and 2)
The drop test of table 1 mixture A
Table 2: the blood pressure lowering experiment of mixture B
Conclusion:
(1) blood pressure is not made significant difference by mixture B.
(2) blood pressure is not made significant difference by the low dose of mixture A, dosage 20mg/Kg 90min upon administration shrink pressure and
Diastolic pressure is significantly less than matched group, and hereafter blood pressure slightly gos up, and dosage 40mg/Kg blood pressure continuous decrease upon administration, from
After 90min, blood pressure reduces amplitude more than 20mmHg.
Comprehensive blood pressure lowering numerical value considers, it can be assumed that mixture A has buck functionality.
Comparative example 2: mixture C, the blood pressure lowering experiment of D
1 prepares sample
(1) mixture C, D redistilled water are configured to three different concentration.
Mixture C: 8mg/ml 4mg/ml 2mg/ml
Mixture D: 8mg/ml 4mg/ml 2mg/ml
(2) set redistilled water and make negative control
2 experimental techniques
Take rat, with 20 urethane anesthesia after weighing.Face upward position to fix, separate common carotid artery, intubate recording blood pressure (instrument
Device: RM6000 tetra-leads physiograph, AP-601, AP-611 recording blood pressure), the diastolic pressure of animal and contraction pressure are direct by instrument
Read.
Post operation stablizes 30min, recording blood pressure.Gavage is dissolved in the mixture C of redistilled water, D, two kinds of mixture dosage
It is respectively 40mg/Kg, 20mg/Kg, 10mg/Kg, additionally sets gavage and give redistilled water and make negative control.Giving mixture C, D
Rear 30min, 60min, 90min, 120min, 150min, 180min recording blood pressure respectively.
3 data process
Calculating average and the standard deviation shrinking pressure and diastolic pressure respectively, between group, difference uses t check analysis.Mixture C with
P ＜ 0.05 between mixture D, there were significant differences.
4 results (see table 3, table 4)
The drop test of table 3 mixture C
The drop test of table 4 mixture D
Conclusion:
(1) blood pressure is not made significant difference by mixture D.
(2) blood pressure is not made significant difference by the low dose of mixture C, dosage 20mg/Kg 90min upon administration shrink pressure and
Diastolic pressure is significantly less than matched group, and hereafter blood pressure slightly gos up, and dosage 40mg/Kg blood pressure continuous decrease upon administration, from
After 90min, blood pressure reduces amplitude more than 20mmHg.
Comprehensive blood pressure lowering numerical value considers, it can be assumed that mixture C has hypotensive effect.
Comparative example 3: mixture E, the blood pressure lowering experiment of F
1 prepares sample
(1) mixture E, F are configured to three different concentration with redistilled water respectively.
Mixture E: 8mg/ml 4mg/ml 2mg/ml
Mixture F:8mg/ml 4mg/ml 2mg/ml
(2) set redistilled water and make negative control
2 experimental techniques
Take rat, with 20 urethane anesthesia after weighing.Face upward position to fix, separate common carotid artery, intubate recording blood pressure (instrument
Device: RM6000 tetra-leads physiograph, AP-601, AP-611 recording blood pressure), the diastolic pressure of animal and contraction pressure are direct by instrument
Read.
Post operation stablizes 30min, recording blood pressure.Gavage is dissolved in the mixture E of redistilled water, F, two kinds of mixture respectively
Dosage is respectively 40mg/Kg, 20mg/Kg, 10mg/Kg, additionally sets gavage and gives redistilled water and make negative control.Mixing respectively
30min after compound E, F, 60min, 90min, 120min, 150min, 180min recording blood pressure respectively.
3 data process
Calculating average and the standard deviation shrinking pressure and diastolic pressure respectively, between group, difference uses t check analysis.Mixture E with
P ＜ 0.05 between mixture F, there were significant differences.
4 results (see table 5, table 6)
(1) blood pressure is not made significant difference by mixture F
(2) blood pressure is not made significant difference by the low dose of mixture E, dosage 20mg/Kg 90min upon administration shrink pressure and
Diastolic pressure is significantly less than matched group, and hereafter blood pressure slightly gos up, and dosage 40mg/Kg blood pressure continuous decrease upon administration, from
After 90min, blood pressure reduces amplitude more than 20mmHg.
Comprehensive blood pressure lowering numerical value considers, it can be assumed that mixture E has hypotensive effect.
The drop test of table 5 mixture E
The blood pressure lowering experiment of table 6 mixture F
The blood pressure lowering experiment of comparative example 4 mixture A1 and B1
1 prepares sample
(1) mixture A1, B1 are configured to three different concentration with redistilled water respectively.
Mixture A1:8mg/ml 4mg/ml 2mg/ml
Mixture B1:8mg/ml 4mg/ml 2mg/ml
(2) set redistilled water and make negative control
2 experimental techniques
Take rat, with 20 urethane anesthesia after weighing.Face upward position to fix, separate common carotid artery, intubate recording blood pressure (instrument
Device: RM6000 tetra-leads physiograph, AP-601, AP-611 recording blood pressure), the diastolic pressure of animal and contraction pressure are direct by instrument
Read.
Post operation stablizes 30min, recording blood pressure.Gavage is dissolved in mixture A1, B1 of redistilled water, two kinds of mixing respectively
Agent amount is respectively 40mg/Kg, 20mg/Kg, 10mg/Kg, additionally sets gavage and gives redistilled water and make negative control.Giving respectively
30min after mixture A1, B1,60min, 90min, 120min, 150min, 180min recording blood pressure respectively.
3 data process
Calculating average and the standard deviation shrinking pressure and diastolic pressure respectively, between group, difference uses t check analysis.Mixture A1 with
P ＜ 0.05 between mixture B1, there were significant differences.
4 results (see table 7, table 8)
(1) blood pressure is not made significant difference by mixture B1
(2) blood pressure is not made significant difference by the low dose of mixture A1, and dosage 20mg/Kg 90min upon administration shrinks pressure
With diastolic pressure significantly less than matched group, hereafter blood pressure slightly gos up, and dosage 40mg/Kg blood pressure continuous decrease upon administration, from
After 90min, blood pressure reduces amplitude more than 20mmHg.
Comprehensive blood pressure lowering numerical value considers, it can be assumed that mixture A1 has hypotensive effect.
The drop test of table 7 mixture A1
The blood pressure lowering experiment of table 8 mixture B1
The blood pressure lowering experiment of comparative example 5 mixture C 1 and D1
1 prepares sample
(1) mixture C 1, D1 redistilled water are configured to three different concentration.
Mixture C 1:8mg/ml 4mg/ml 2mg/ml
Mixture D 1:8mg/ml 4mg/ml 2mg/ml
(2) set redistilled water and make negative control
2 experimental techniques
Take rat, with 20 urethane anesthesia after weighing.Face upward position to fix, separate common carotid artery, intubate recording blood pressure (instrument
Device: RM6000 tetra-leads physiograph, AP-601, AP-611 recording blood pressure), the diastolic pressure of animal and contraction pressure are direct by instrument
Read.
Post operation stablizes 30min, recording blood pressure.Gavage is dissolved in the mixture C 1 of redistilled water, D1, two kinds of mixing respectively
Agent amount is respectively 40mg/Kg, 20mg/Kg, 10mg/Kg, additionally sets gavage and gives redistilled water and make negative control.Mixing
30min after thing A, B, 60min, 90min, 120min, 150min, 180min recording blood pressure respectively.
3 data process
Calculating average and the standard deviation shrinking pressure and diastolic pressure respectively, between group, difference uses t check analysis.Mixture C 1 with
P ＜ 0.05 between mixture D 1, there were significant differences.
4 results (see table 9 and 10)
The drop test of table 9 mixture C 1
The blood pressure lowering experiment of table 10 mixture D 1
Conclusion:
(1) blood pressure is not made significant difference by mixture D 1.
(2) blood pressure is not made significant difference by the low dose of mixture C 1, and dosage 20mg/Kg 90min upon administration shrinks pressure
With diastolic pressure significantly less than matched group, hereafter blood pressure slightly gos up, and dosage 40mg/Kg blood pressure continuous decrease upon administration, from
After 90min, blood pressure reduces amplitude more than 20mmHg.
Comprehensive blood pressure lowering numerical value considers, it can be assumed that mixture A has buck functionality.
The antihypertensive activity experiment of comparative example 6 Lumbricus powder
One: the preparation of Lumbricus powder
1, fresh William's cavity ring earthworm is taken standby through telling mud.
2, water cleans.With baking oven, under the temperature conditions of 40～60 DEG C, dry 10 to 20 hours, standby.
3, the Lumbricus direct mechanical dried is smashed to pieces powdering, standby.
Two: the antihypertensive activity experiment of above-mentioned Lumbricus powder (hereinafter referred to as Lumbricus powder)
1 prepares sample
(1) Lumbricus powder redistilled water is configured to three different concentration.
Lumbricus powder: 1.2g/ml 0.6g/ml 0.3g/ml
(2) set redistilled water and make negative control
2 experimental techniques
Take rat, with 20% urethane anesthesia after weighing.Face upward position to fix, separate common carotid artery, intubate recording blood pressure (instrument:
RM6000 tetra-leads physiograph, AP-601, AP-611 recording blood pressure), the diastolic pressure of animal and contraction pressure are directly read by instrument
Go out.
Post operation stablizes 30min, recording blood pressure.Gavage is dissolved in redistilled water Lumbricus powder, and dosage is 6g/Kg, 3g/Kg,
1.5g/Kg, additionally sets gavage and gives redistilled water and make negative control.30min, 60min, 90min after giving Lumbricus powder,
120min, 150min, 180min recording blood pressure respectively.
3 data process
With molecular weight less than 3000 daltonian William's cavity ring earthworm extract results contrast (" mixing in previous embodiment 2
Thing A ").Calculating average and the standard deviation shrinking pressure and diastolic pressure respectively, between group, difference uses t check analysis.P ＜ between two groups
0.01, there is pole significant difference.
4 results (be shown in Table 11, table 12)
The drop test of table 11 mixture A
Table 12 Lumbricus powder drop test
Conclusion:
(1) blood pressure is had no significant effect by the dosage of the 1.5g/Kg of Lumbricus powder and the dosage of 3g/Kg, and the dosage of 6g/Kg can make
Blood pressure drops, but not statistically significant.
(2) blood pressure is not made significant difference by the low dose of mixture A, dosage 20mg/Kg 90min upon administration shrink pressure and
Diastolic pressure is significantly less than matched group, and hereafter blood pressure slightly gos up, and dosage 40mg/Kg blood pressure continuous decrease upon administration, from
After 90min, blood pressure reduces amplitude more than 20mmHg.
Comprehensive blood pressure lowering numerical value considers, it can be assumed that mixture A has buck functionality, its activity is significantly better than Lumbricus powder.
Claims (9)
1. it is used for blood pressure lowering or treatment or the earthworm extract of prophylaxis of hypertension, it is characterised in that: described earthworm extract is
Molecular weight is the water extract of the Lumbricus of below 3000 dalton, and described Lumbricus is William's cavity ring earthworm, Eisenia foetida and saw
One or more in the remote earthworm of tooth, described earthworm extract is to be prepared by the preparation method comprised the steps of: by Lumbricus
Water extract, through 3000 molecular weight ultrafiltration through membranes, collects filtrate,.
2. the earthworm extract described in claim 1, the water extract of wherein said Lumbricus is prepared by following method: will
Lumbricus after telling mud, washing process smashs pulp to pieces, soaks after the mixing that adds water, then with the centrifuge of 1000-3000 rev/min
Centrifugal, obtain supernatant 1, by supernatant 1 with the centrifuge of 10000-20000 rev/min, obtain supernatant 2, obtaining supernatant
Drying steps is also comprised after 2.
3. the earthworm extract described in claim 2, wherein, the described amount added water is 0.8-1.5 times of pulpous state Lumbricus volume；
The time of described immersion is 3-10 hour；The temperature of described immersion is 4-10 DEG C.
4. the earthworm extract described in claim 1, wherein, was also including following step before 3000 molecular weight ultrafiltration through membranes
Rapid: after the water extract of Lumbricus is dissolved in water, then be centrifuged with 10000-20000 rev/min with centrifuge, obtain supernatant；Wherein
The water extract of described Lumbricus is dried object.
5. the earthworm extract described in claim 4, the 1-5 of the wherein said water extract volume that the amount added water is Lumbricus
Times.
6. the earthworm extract described in claim 1, wherein said Lumbricus is William's cavity ring earthworm.
7. the earthworm extract described in claim 1, wherein said Lumbricus is Eisenia foetida.
8. the earthworm extract described in claim 1, wherein said Lumbricus is Amythas dancatala.
9. arbitrary described earthworm extract of claim 1-8 is preparing the medicine of blood pressure lowering or in preparation treatment or prevention
Application in the health food of hypertension.
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