CN105535048A - Application of celery seed extract to preparation of medicine or health-care food for resisting to hyperuricemia and gout - Google Patents
Application of celery seed extract to preparation of medicine or health-care food for resisting to hyperuricemia and gout Download PDFInfo
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- CN105535048A CN105535048A CN201610003095.1A CN201610003095A CN105535048A CN 105535048 A CN105535048 A CN 105535048A CN 201610003095 A CN201610003095 A CN 201610003095A CN 105535048 A CN105535048 A CN 105535048A
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- China
- Prior art keywords
- apium graveolens
- hyperuricemia
- gout
- linnaeus extract
- graveolens linnaeus
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
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Abstract
The invention relates to application of celery seed extract to preparation of medicine or health-care food for resisting to hyperuricemia and gout and further provides medicine or health-care food for treating hyperuricemia and gout. Celery seed extract serves as the active constituent, and an appropriate number of medical carriers or auxiliary constituents are added, so that a preparation is obtained. The medicine for health-care food is simple in preparation process, the celery seed extract can obviously lower serum uric acid of model mice suffering from hyperuricemia caused by oteracil potassium salt, lower the activity of xanthine oxidase to different degrees, achieve an obvious inhibition effect on arthritis of the mice, prevent and treat hyperuricemia or gout and be applied to preparation of the medicine or health-care food for resisting to hyperuricemia and gout.
Description
Technical field
The present invention relates to Natural Medicine Chemistry and medical art, specifically, is the application of a kind of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout, and the application in the health food preparing anti-hyperuricemia and anti-gout drugs.
Background technology
In traditional Chinese medical science tradition ancient books and records are recorded, the category that gout belongs to " arthromyodynia ", the cause of disease is many to cause damp-heat accumulation, damp and hot resistance network with intemperance of taking food, addicted to eat sweet plump taste, accumulates and hold back heat-transformation, show as pain very, unrestrained to swell very, the symptom such as thermogravimetric.Gout is long-term disorders of purine metabolism, and blood uric acid continues to increase and causes urate crystal to deposit, and causes one group of clinical syndrome of the tissue injurys such as characteristic chronic arthritis and joint deformity.Improving constantly in recent years along with people's living standard, dietary structure also there occurs change, too much absorption purine and protein, make the sickness rate of China's gout in rising year by year, age of onset presents and becomes younger, and brings huge burden to society, and it is closely related with the generation of the diseases such as hypertension, hyperlipemia, diabetes, become the early sign identifying metabolism syndrome, gout is the independent hazard factor of All-cause death and cardiovascular death, which significantly reduces the life quality of patient.
For the treatment of goat, except carrying out diet control, the more drug main of use will based on Western medicine at present, comprise antiinflammatory class medicine to have: colchicine, nonsteroidal antiinflammatory drug (NSAIDs), glucocorticoid, adrenocortical hormone, glucosamine, IL-1 inhibitor etc.; Uric acid resisting class medicine has: promote urate excretion medicine and suppress uricopoiesis medicine.Wherein allopurinol is a kind of xanthine oxidase inhibitor, the synthesis of purine nucleotides is reduced by the enzyme two kinds of approach consuming phosphoribosylpyrophosphate synthetase (PRPP) and feedback suppression purine nucleotides de novo synthesis, reach minimizing uricopoiesis, reduce the effect of blood and urine uric acid level.But Western medicine shows it mostly has more serious untoward reaction, such as hepatorenal damage, super quick allergy, bone marrow depression etc., limit its safe handling.Research finds, Chinese medicine also has good effect to treatment gout, and display is less or the toxic and side effects of nothing.Some Chinese medicines are by antiinflammatory, analgesic activities foot swelling inhibition and playing a role, as liriope muscari Baily, Alcohol extractive of qinjiao, Cortex Phellodendri etc., TianYQ etc., find in liriope muscari Baily water extract and the research of effective site anti-inflammatory activity thereof, adopt dimethylbenzene inducing mouse auricle edema and carrageenin or histamine inducing mouse pedal swelling model, liriope muscari Baily has remarkable inside and outside anti-inflammatory activity.Other Chinese medicine plays a role, as Cortex Cinnamomi withe, Caulis Gneti, Herba Lycopi, Rhizoma Polygoni Cuspidati water extract etc. by suppressing the activity of XOD.The water extract of Cortex Phellodendri and ERMIAO PILL and the hyperuricemia model mice of aesculin to Oteracil Potassium Salt treatment have obvious uric acid resisting effect.
Celery seed is the seed of Herba Apii graveolentis, Herba Apii graveolentis not only has higher edibility, its fruit also containing abundant nutrient and the multiple material with physiologically active, mainly comprises volatile oil compound, unsaturated fatty acid, flavonoid substances, butyl benzene phthaleins, aminoacid, dietary fiber, coumarin derivative etc.Celery seed has higher edible and medical value, its functional components has the biological activitys such as good antiinflammatory, antioxidation, antitumor, antibacterial and parasite killing, diuretic as a kind of effective treatment cystitis and nephropathy is applied clinically, in addition, be also widely used in fields such as food, medicine, cosmetics.
Celery seed treatment arthralgia has century-old history in Australia, be regarded as Traditional Folk folk prescription, record according to nineteen eighty-three version British Herbal Pharmacopoeia, celery seed has the effect of the complication such as treatment gout, pertinent literature is reported, Apium graveolens Linnaeus extract has obvious prevention and cure of cardiovascular disease, suppresses the pharmacological action of cancer growth rate.Containing the nutrition useful to articular cartilage in modern nutriology determination seed oil of Herba Apii graveolentis.Apium graveolens Linnaeus extract not only antiphlogistic effects is good; and can significantly reduce hyperuricemia level; there is obviously anti-antipodagric effect; there is certain protective effect to the renal function injury that gouty arthritis produces simultaneously; Apium graveolens Linnaeus extract is a kind of effective ingredient extracted from celery seed, and its main active is flavones ingredient and the butylphthalide compounds such as chrysoeriol, luteolin and apigenin.Research shows, chrysoeriol, luteolin and apigenin significantly can suppress the activity of xanthine oxidase in vitro, and it is active suitable with the activity of allopurinol.In addition, butylphthalide compounds is the compound that a class has sedation-analgesia effect.So far, toxicology data display celery seed there is no obvious toxic and side effects.Therefore celery seed is as anti-antipodagric new Chinese medicine research and development, and tool has great advantage and good prospect.
Celery seed has the effect for the treatment of gout and complication thereof, etc. PowandaM.C. research proves that the diuresis that celery seed is good and antiinflammatory action can be used as treatment rheumatism and arthritic adjuvant drug (PowandaM.C., RainsfordK.D.Atoxicologicalinvestigationofaceleryseedext racthavinganti-inflammatoryactivity.Inflammopharmacology, 2010,19 (4): 227-233.); Tong Guohui etc. extract on the impact of hyperuricemia rat blood serum uric acid level and for experimental study analgesic effect celery seed, result shows that Apium graveolens Linnaeus extract has certain protective effect (Tong Guohui to hyperuricemia and the renal function injury that causes thereof, Deng, Apium graveolens Linnaeus extract is on the impact of hyperuricemia rat, Food Science, 2008,29 (6): 641-644.); Patent ZL200710062800.6 obtains celery seed acetic acid ethyl ester extract to Apium graveolens Linnaeus extract after petroleum ether and ethyl acetate repeatedly extract, contained extract contains 6 Coumarinses and 12 flavone compounds, only xanthine oxidase vitro inhibition activity is carried out to the chrysoeriol in 18 compounds, luteolin, apigenin, chrysoeriol is on the impact of hyperuricemia model on mice, and celery seed acetic acid ethyl ester extract is to the experiment of mice foot swelling suppression ratio, but foot swelling suppression ratio is only 10.1% (positive controls suppression ratio 24.9%).In correlational study paper, in celery seed acetic acid ethyl ester extract, chrysoeriol, luteolin, Quantitative Determination of Apigenin are only 0.148mg/g, 0.176mg/g, 0.145mg/g (Xu Na, celery seed active component and anti-gout novel medical research thereof, The 2nd Army Medical College master thesis, 2012), pharmacologically active effect content of monomer is low and acetic acid ethyl ester extract is only 10.1% to mice foot swelling suppression ratio.Patent ZL20100615591.2 protects the oral liquid of celery seed ethanol extraction treatment gout or soft capsule preparation, but not yet carries out relevant pharmacology Study on mechanism.
Summary of the invention
The technical problem to be solved in the present invention is the application of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food.The animal model of the present invention to the anti-hyperuricemia of celery seed ethanol extraction and gout is tested, its extract causes mice foot swelling model by Carrageenan and uric acid, observes the therapeutical effect of Apium graveolens Linnaeus extract to hyperuricemia and gouty arthritis.Experiment shows that Apium graveolens Linnaeus extract obviously can reduce the serum uric acid level of hyperuricemia mice, and its uric acid resisting effect is relevant with suppressing the activity of xanthine oxidase, and Apium graveolens Linnaeus extract has obvious inhibitory action to mouse arthritis.Illustrate that Apium graveolens Linnaeus extract has good preventive and therapeutic action in anti-hyperuricemia and gout.
In order to realize above-mentioned purpose of the present invention, present invention employs following technical scheme:
The application of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, described medicine take Apium graveolens Linnaeus extract as active component, and Apium graveolens Linnaeus extract content accounts for 10 ~ 100% of active constituents of medicine quality.
According to the application of described a kind of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, in wherein said Apium graveolens Linnaeus extract, GraveobiosideA content is not less than 9%.Described Apium graveolens Linnaeus extract is obtained by following methods: get Herba Apii graveolentis dry seed, with 8 ~ 15 times amount 30 ~ 70%V/V ethanol percolate extraction or heating extraction 2 ~ 4 times, and each 1.5 ~ 3 hours, extracting solution concentrating under reduced pressure, dry.
According to the application of described a kind of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, wherein said application is the xanthine oxidase activity that Apium graveolens Linnaeus extract passes through to reduce serum, reduce serum uric acid level, reach uric acid resisting effect.
According to the application of described a kind of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, wherein said application is that Apium graveolens Linnaeus extract passes through to suppress foot swelling caused by Carrageenan, reaches prevention gouty arthritis foot swelling effect.
According to the application of described a kind of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, wherein said application is by suppressing foot swelling caused by uric acid, reaches prevention gouty arthritis foot swelling effect.
According to the application of described a kind of Apium graveolens Linnaeus extract in the anti-hyperuricemia of preparation and the medicine of gout or health food, wherein said application is Apium graveolens Linnaeus extract concentration when being 200 μ g/mL to external xanthine oxidase suppression ratio is 45.32 ± 8.94% (x ± sd).
According to the application of described a kind of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, the pharmaceutical preparation of making for active component with 0.5g ~ 6.0g Apium graveolens Linnaeus extract is taken in wherein said application for each person every day.
Apium graveolens Linnaeus extract, is obtained by following methods: get Herba Apii graveolentis dry seed, with 8 ~ 15 times amount 30 ~ 70%V/V ethanol percolate extraction or heating extraction 2 ~ 4 times, and each 1.5 ~ 3 hours, extracting solution concentrating under reduced pressure, dry.
The pharmaceutical composition of anti-hyperuricemia and gout, containing Apium graveolens Linnaeus extract and pharmaceutically suitable carrier and/or excipient, described Apium graveolens Linnaeus extract is obtained by following methods: get Herba Apii graveolentis dry seed, with 8 ~ 15 times amount 30 ~ 70%V/V ethanol percolate extraction or heating extraction 2 ~ 4 times, each 1.5 ~ 3 hours, extracting solution concentrating under reduced pressure, dry; Apium graveolens Linnaeus extract content accounts for 10 ~ 100% of active constituents of medicine quality; In Apium graveolens Linnaeus extract, the content of GraveobiosideA is 8 ~ 11%.
Apium graveolens Linnaeus extract of the present invention is through following zoopery and the experiment of external xanthine oxidase suppression ratio, prove that Apium graveolens Linnaeus extract obviously can reduce the serum uric acid level of hyperuricemia mice, its uric acid resisting effect is relevant with suppressing the activity of xanthine oxidase, Apium graveolens Linnaeus extract is the competitive inhibitor of xanthine oxidase, has obvious inhibitory action to mouse arthritis simultaneously.
By preparing the salt-induced Studies on Animal Models of Hyperuricemic Mice of Oteracil Potassium, observe Apium graveolens Linnaeus extract to the preventive effect of hyperuricemia.Basic, normal, high three the dosage groups (being followed successively by 50mg/kg, 100mg/kg, 200mg/kg by body weight dose group) of Apium graveolens Linnaeus extract are compared with model control group, Normal group, positive controls (medicine is allopurinol 40mg/kg), compared with Normal group, model group animal serum uric acid and xanthine oxidase level significantly raise, and show Studies on Animal Models of Hyperuricemic Mice success; All administration groups all obviously can reduce the serum uric acid level of the salt-induced hyperuricemia mice of Oteracil Potassium, significant difference compared with model control group; Wherein, positive control allopurinol, Apium graveolens Linnaeus extract 200mg/kg group also obviously reduce the xanthine oxidase activity of model mice serum.Result shows that Apium graveolens Linnaeus extract obviously can reduce the serum uric acid level of hyperuricemia mice, and its uric acid resisting effect is relevant with suppressing the activity of xanthine oxidase.
Cause mice foot swelling model by preparation on Carrageenan, observe Apium graveolens Linnaeus extract to the preventive effect of gouty arthritis.Basic, normal, high three the dosage groups (being followed successively by 50mg/kg, 100mg/kg, 200mg/kg by body weight dose group) of Apium graveolens Linnaeus extract are compared with model control group and positive controls (medicine is indometacin tablets 10mg/kg), and Apium graveolens Linnaeus extract, positive control Indomethacin's sheet all obviously can suppress the foot swelling of on Carrageenan induced mice.Result shows that Apium graveolens Linnaeus extract has obvious preventive effect to gouty arthritis foot swelling.
Causing mice foot swelling model by preparing uric acid, observing Apium graveolens Linnaeus extract to the preventive effect of gouty arthritis.Basic, normal, high three the dosage groups (being followed successively by 50mg/kg, 100mg/kg, 200mg/kg by body weight dose group) of Apium graveolens Linnaeus extract and model control group and positive controls (medicine be indometacin tablets 10mg/kg, colchicines tablets 1.0mg/kg) compare, only have except Apium graveolens Linnaeus extract low dose group outside effect trend, positive control Indomethacin's sheet, colchicines tablets, Apium graveolens Linnaeus extract senior middle school dosage all obviously can suppress the foot swelling of uric acid induced mice.Result shows that Apium graveolens Linnaeus extract has obvious preventive effect to gouty arthritis foot swelling.
By testing the external xanthine oxidase suppression ratio of Apium graveolens Linnaeus extract, take allopurinol as positive control, Apium graveolens Linnaeus extract concentration is under 200 μ g/mL, and measure the suppression ratio of xanthine oxidase, test result is 45.32 ± 8.94% (x ± sd).
Apium graveolens Linnaeus extract of the present invention, the content of GraveobiosideA is detected by HPLC, chromatographic condition is Shimadzu LC-2010AHT high performance liquid chromatograph, SHIMADZU (5 μm, VP-ODS, 150L × 4.6) chromatographic column, determined wavelength 345nm, flow velocity is 0.8ml/min, column temperature 30 DEG C, mobile phase is acetonitrile: 0.05% aqueous formic acid (19:81), the GraveobiosideA in four parallel assay Apium graveolens Linnaeus extract, and the content calculating GraveobiosideA according to external standard method is 9.5%.
Apium graveolens Linnaeus extract of the present invention is when preparing anti-hyperuricemia and anti-gout drugs or health food, and Apium graveolens Linnaeus extract content accounts for 10 ~ 100% of active component quality.
Apium graveolens Linnaeus extract of the present invention is used for the treatment of hyperuricemia or gout, is the preparation made that to take every day with 0.5g ~ 6.0g Apium graveolens Linnaeus extract be active component.
Described preparation is the various dosage forms that can use clinically, as capsule, granule, pill, tablet, unguentum, tincture, oral liquid; Or existing health food dosage form, as capsule, granule, tablet, beverage etc.
The invention provides the application of Apium graveolens Linnaeus extract in the anti-hyperuricemia of preparation and anti-gout drugs or health food, expand the scope of resource of anti-hyperuricemia and anti-gout drugs or health food.
Compared with prior art, the present invention possesses following excellent benefit:
In Apium graveolens Linnaeus extract of the present invention, GraveobiosideA content is 9.5%; Apium graveolens Linnaeus extract is to the obvious preventive effect of gouty arthritis foot swelling, and causing mice foot swelling suppression ratio to uric acid is 31.5%, and causing mice foot swelling suppression ratio on Carrageenan is 29.7%; Apium graveolens Linnaeus extract obviously reduces the serum uric acid level of the salt-induced hyperuricemia mice of Oteracil Potassium, Apium graveolens Linnaeus extract 200mg/kg dosage group Mouse Blood uric acid is 133.0 μm of ol/L, rate of change 32.77%, also obviously reduce the xanthine oxidase activity of model mice serum, Apium graveolens Linnaeus extract 200mg/kg dosage group Mouse Blood xanthine oxidase is 644.0U/L, and rate of change is 12.72%; Apium graveolens Linnaeus extract is external is 45.32% to xanthine oxidase suppression ratio, has significant inhibit activities.Show that Apium graveolens Linnaeus extract has good preventive and therapeutic action in anti-hyperuricemia and gout.
Through patent and literature search, the present invention and patent ZL200710062800.6, ZL20100615591.2 have certain similarity, but patent ZL200710062800.6 mainly carries out relevant pharmacology activity experiment to the monomer in celery seed acetic acid ethyl ester extract, but not to the pharmacology activity research that celery seed ethanol extraction of the present invention is correlated with.The compositions of patent ZL20100615591.2 to celery seed ethanol extraction is protected, but not yet carries out relevant pharmacology Study on mechanism.The present invention is directed to that celery seed 70% ethanol extraction has carried out the experiment of Oteracil Potassium salt-induced mouse hyperuricemia, on Carrageenan, uric acid cause the related experiment such as mice foot swelling experiment and the experiment of external xanthine oxidase suppression ratio; prove that Apium graveolens Linnaeus extract has good preventive and therapeutic action in anti-hyperuricemia and gout, from patent ZL200710062800.6, ZL20100615591.2, there is different protection content.
Accompanying drawing illustrates:
Fig. 1 is the structural formula schematic diagram of GraveobiosideA;
Fig. 2 is the HPLC figure of GraveobiosideA;
The HPLC figure of Fig. 3 Apium graveolens Linnaeus extract.
Detailed description of the invention:
Below in conjunction with accompanying drawing, further illustrate essentiality content of the present invention with embodiments of the invention, but do not limit the present invention with this.
Embodiment 1:
The preparation of Apium graveolens Linnaeus extract:
Herba Apii graveolentis dry seed 20kg, adds 8 times amount 70% (V/V) ethanol heating extraction 2 times, each 2 hours; Merge extractive liquid, extracting solution 65 DEG C of concentrating under reduced pressure, drying obtains Apium graveolens Linnaeus extract 3.2kg.
Embodiment 2:
Apium graveolens Linnaeus extract is on the impact of the salt-induced mouse hyperuricemia of Oteracil Potassium:
The Apium graveolens Linnaeus extract of Example 1, high, medium and low three dosage component are not 200,100,50mg/kg, with pure water be mixed with 10,5, the suspension of 2.5mg/mL concentration; Allopurinol tablet pure water is mixed with the suspension of 2mg/mL concentration.
Get 21 ~ 24g male ICR mouse 72, be divided into 6 groups at random by body weight: Normal group; Model control group; Positive control Allopurinol Tablets 40mg/kg group; Apium graveolens Linnaeus extract 200,100,50mg/kg group.Except positive control allopurinol tablet in experimental day gastric infusion once except, all the other each treated animals equal every days according to dosage gastric infusion once, for three days on end.Mice fasting in 2nd day be can't help water and is spent the night (12h), and morning next day, all the other each treated animals were all by 300mg/kg dosage lumbar injection oxonic acid potassium salt 0.2mL/10g.bw except Normal group injection normal saline.1h last administration after moulding, blood is got in endocanthion after interval 1h (moulding 2h), the centrifugal 10min of 3000r/min, measures each group of Mouse Blood uric acid (UA) and xanthine oxidase (XOD) level by kit method (phosphorus ursolic acid reducing process).
Basic, normal, high dosage Apium graveolens Linnaeus extract group and the impact of allopurinol on hyperuricemia mice uric acid level the results are shown in Table 1, table 2.Compared with Normal group, model group animal serum uric acid and xanthine oxidase level significantly raise, and show Studies on Animal Models of Hyperuricemic Mice success; All administration groups all obviously can reduce the serum uric acid level of the salt-induced hyperuricemia mice of Oteracil Potassium, significant difference compared with model control group; Wherein, positive control allopurinol, Apium graveolens Linnaeus extract 200mg/kg also obviously reduce the xanthine oxidase activity of model mice serum.
The impact that table 1 raises the salt-induced mice UA of Oteracil Potassium
Compared with normal group,
##p<0.01; Compared with model group, */* * P<0.05/0.01
The impact that table 2 raises the salt-induced mice XOD of Oteracil Potassium
Compared with normal group,
##p<0.01; Compared with model group, */* * P<0.05/0.01
Embodiment 3:
Apium graveolens Linnaeus extract causes the impact of mice foot swelling on Carrageenan:
The Apium graveolens Linnaeus extract of embodiment 1, high, medium and low three dosage component are not 200,100,50mg/kg, with pure water be mixed with 10,5, the suspension of 2.5mg/mL concentration; Indometacin enteric-coated tablet pure water is mixed with the suspension of 0.5mg/mL concentration.
Select 19 ~ 21g male ICR mouse, be divided into 5 groups at random by body weight: matched group; Positive control Indomethacin's sheet 10mg/kg, Apium graveolens Linnaeus extract 200,100,50mg/kg group, often organize 12.Except indometacin tablets only in experimental day gastric infusion once except, all the other each treated animals equal every days according to dosage gastric infusion once, for three days on end, matched group gives the pure water of 20mL/kg.bw.After last administration 30 minutes, in each group of right back sufficient plantar subcutaneous injection 1% carrageenin 0.05mL/ foot of mice.Cause scorching after 4h take off cervical vertebra and put to death animal, cut by profit and take off biped in ankle and weigh, using the difference of two heavy sensation in the foot amounts as swelling.
Basic, normal, high dosage Apium graveolens Linnaeus extract group and indometacin tablets the results are shown in Table 3 to the impact that fork dish glue causes mice foot swelling.Result shows that all administration groups all obviously can suppress the foot swelling of carrageenin induced mice, and difference all has statistical significance compared with Vehicle controls group.
Table 3 on Carrageenan causes the impact of mice foot swelling
Compared with matched group, */* * P<0.05/0.01
Embodiment 4:
Apium graveolens Linnaeus extract causes the impact of mice foot swelling to uric acid:
The Apium graveolens Linnaeus extract of embodiment 1, high, medium and low three dosage component are not 200,100,50mg/kg, with pure water be mixed with 10,5, the suspension of 2.5mg/mL concentration; Indometacin enteric-coated tablet pure water is mixed with the suspension of 0.5mg/mL concentration; Colchicines tablets pure water is mixed with the suspension of 0.05mg/mL concentration.The preparation of Monosodium urate: get 194mL distilled water and add 6mL1mol/LNaOH, boil, add 1g uric acid, adjust pH value to 7.2 with 1mol/LHcl, stir cooling, 4 DEG C of Refrigerator store 24h, remove supernatant, blotted by precipitate moisture with filter paper, put into drying baker, 70 DEG C are dried 2h, take out, scrape powder, put into mortar pulverization, bottle for subsequent use.Face the used time is made into 20mg/mL suspension with 1% tween 80, during injection, jolting is even.
20 ~ 22g male ICR mouse 60 is got in experiment, is divided into 6 groups at random: matched group by body weight; Positive control Indomethacin's sheet 10mg/kg, colchicines tablets 1.0mg/kg, Apium graveolens Linnaeus extract 200,100,50mg/kg group; Often organize 10.Except indomethacin and colchicine only in experimental day gastric infusion once except, all the other each treated animals equal every days according to dosage gastric infusion once, for three days on end, Vehicle controls group gives the pure water of 20mL/kg.bw.After last administration 30 minutes, make Monosodium urate suspension 0.05mL/ only (1mg/ only) by oneself in the left toes of mice to ankle joint direction subcutaneous injection.Cause scorching after 5h take off cervical vertebra and put to death animal, cut by profit and take off biped in ankle and weigh, using the difference of two heavy sensation in the foot amounts as swelling.For reducing systematic error, test adopts operation repetitive.
Basic, normal, high dosage Apium graveolens Linnaeus extract group and indometacin tablets, colchicine the results are shown in Table 4 to impact uric acid being caused to mice foot swelling.From table 4, only have except Apium graveolens Linnaeus extract low dose group outside effect trend, positive control Indomethacin's sheet 10mg/kg, colchicines tablets 1.0mg/kg, Apium graveolens Linnaeus extract senior middle school dosage group all obviously can suppress the foot swelling of uric acid induced mice, all have statistical significance with difference compared with solvent photograph group.
Table 4 causes the impact of mice foot swelling to Monosodium urate
Compared with matched group,
*p<0.01
Embodiment 5:
The external xanthine oxidase suppression ratio experiment of Apium graveolens Linnaeus extract:
The Apium graveolens Linnaeus extract of embodiment 1, gets 1.00mg, dissolves and is diluted to 1.0mL with buffer, obtain 1.00mg/mL mother solution with 20 μ LDMSO.Through following steps, the final concentration of sample test is 200.00 μ g/ml.100 μ L sodium pyrophosphate buffer solutions (pH=7.4) are added in blank group; Negative control group first adds 60 μ L buffer; 20 μ L buffer and 40 μ L enzymatic solution (2.5U/L) are added in enzyme group; Sample determination group adds 20 μ L samples and 40 μ L enzymatic solution (2.5U/L); Sample Negative matched group adds 20 μ L sample solutions and 40 μ L buffer; 20 μ L allopurinol solution (50 μMs) and 40 μ L enzymatic solution (2.5U/L) are added in positive controls.After application of sample completes, put it in calorstat and hatch 15 minutes at 25 DEG C.After having hatched, except blank group, all the other all add 40 μ L xanthine solution (400 μMs), the final volume often organized is 0.1mL, and carries out dynamic method mensuration at 295nm wavelength immediately, and the reading time is 30 minutes, 5 minutes/time, parallel assay three times.
Xanthine oxidase suppression ratio computing formula: (△ enzyme-△ sample/△ enzyme-△ is cloudy) × 100%, active strong and weak by calculating the xanthine oxidase suppression ratio detecting given the test agent, Apium graveolens Linnaeus extract is 45.32% ± 8.94% (x ± sd) to xanthine oxidase suppression ratio, has significant inhibit activities.
Embodiment 6:
HPLC detects the content of GraveobiosideA in Apium graveolens Linnaeus extract:
Chromatographic condition is Shimadzu LC-2010AHT high performance liquid chromatograph, and SHIMADZU (5 μm, VP-ODS, 150L × 4.6) chromatographic column, determined wavelength 345nm, flow velocity is 0.8ml/min, column temperature 30 DEG C, mobile phase is acetonitrile: 0.05% aqueous formic acid (19:81).
Reference substance is prepared: precision takes reference substance GraveobiosideA5.00mg, is placed in the volumetric flask of 10ml, and the EtOH Sonicate with 50% dissolves, and is settled to scale, with 0.45 μm of membrane filtration.Test sample is prepared: precision takes the Apium graveolens Linnaeus extract 25.00mg of embodiment 1, is placed in the volumetric flask of 10ml, and the EtOH Sonicate with 50% dissolves, and is settled to scale, with 0.45 μm of membrane filtration.
GraveobiosideA structural formula is shown in Fig. 1, and GraveobiosideA reference substance and Apium graveolens Linnaeus extract chromatogram are shown in Fig. 2, Fig. 3.GraveobiosideA in four parallel assay Apium graveolens Linnaeus extract, calculate content according to external standard method, the peak area of GraveobiosideA reference substance is 5345953, and the peak area of impurity peaks is 205175, and its purity is 96.30%.In Apium graveolens Linnaeus extract, GraveobiosideA tetra-detected peaks areas are 2532486,2439593,2256169,2228582, meansigma methods is 2364207 (RSD%), and GraveobiosideA content meansigma methods is 9.519 ± 0.28% (± sd).
Embodiment 7:
First obtain Apium graveolens Linnaeus extract by the mode of embodiment 1, mix homogeneously with the ratio of excipient weight than 4:1 in it, granulate, tabletting.Treatment hyperuricemia and gout 3 ~ 5 pieces/times, 3 times/day.
Embodiment 8:
First obtain Apium graveolens Linnaeus extract by the mode of embodiment 1, mix homogeneously with the ratio of excipient weight than 8:1 in it, granulate, tabletting.Treatment hyperuricemia and gout 2 ~ 4 pieces/times, 2 times/day.
Embodiment 9:
First obtain Apium graveolens Linnaeus extract by the mode of embodiment 1, mix homogeneously with the ratio of excipient weight than 3:1 in it, granulate, make capsule.Treatment hyperuricemia and gout 3 ~ 6 tablets/time, 3 times/day.
Embodiment 10:
First obtain Apium graveolens Linnaeus extract by the mode of embodiment 1, mix homogeneously with the ratio of excipient weight than 6:1 in it, granulate, make capsule.Treatment hyperuricemia and gout 2 ~ 4 tablets/time, 3 times/day.
Embodiment 11:
First obtain Apium graveolens Linnaeus extract by the mode of embodiment 1, the method for making of oral liquid makes oral liquid routinely.Treatment hyperuricemia and gout 5 ~ 20ml tablet/time, 2-3 times/day.
Claims (9)
1. the application of Apium graveolens Linnaeus extract in the medicine preparing anti-hyperuricemia and gout or health food, it is characterized in that, described medicine take Apium graveolens Linnaeus extract as active component, and Apium graveolens Linnaeus extract content accounts for 10 ~ 100% of active constituents of medicine quality.
2. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the medicine preparing anti-hyperuricemia and gout or health food, it is characterized in that, in described Apium graveolens Linnaeus extract, GraveobiosideA content is 8 ~ 11%.
3. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the medicine preparing anti-hyperuricemia and gout or health food, it is characterized in that, described Apium graveolens Linnaeus extract is obtained by following methods: get Herba Apii graveolentis dry seed, with 8 ~ 15 times amount 30 ~ 70%V/V ethanol percolate extraction or heating extraction 2 ~ 4 times, each 1.5 ~ 3 hours, extracting solution concentrating under reduced pressure, dry.
4. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the medicine preparing anti-hyperuricemia and gout or health food, it is characterized in that described application is the xanthine oxidase activity that Apium graveolens Linnaeus extract passes through to reduce serum, reduce serum uric acid level, reach uric acid resisting effect.
5. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the medicine preparing anti-hyperuricemia and gout or health food, it is characterized in that described application is that Apium graveolens Linnaeus extract passes through to suppress foot swelling caused by Carrageenan, reach prevention gouty arthritis foot swelling effect.
6. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the medicine preparing anti-hyperuricemia and gout or health food, it is characterized in that described application is by suppressing foot swelling caused by uric acid, reaching prevention gouty arthritis foot swelling effect.
7. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the anti-hyperuricemia of preparation and the medicine of gout or health food is 45.32 ± 8.94% (x ± sd) to external xanthine oxidase suppression ratio when it is characterized in that described application be Apium graveolens Linnaeus extract concentration is 200 μ g/mL.
8. the application of a kind of Apium graveolens Linnaeus extract according to claim 1 in the medicine preparing anti-hyperuricemia and gout or health food, is characterized in that the pharmaceutical preparation of making for active component with 0.5g ~ 6.0g Apium graveolens Linnaeus extract is taken in described application for each person every day.
9. the pharmaceutical composition of anti-hyperuricemia and gout, containing Apium graveolens Linnaeus extract and pharmaceutically suitable carrier or excipient, in described Apium graveolens Linnaeus extract, the content of GraveobiosideA is 8 ~ 11%, obtained by following methods: get Herba Apii graveolentis dry seed, with 8 ~ 15 times amount 30 ~ 70%V/V ethanol percolate extraction or heating extraction 2 ~ 4 times, each 1.5 ~ 3 hours, extracting solution concentrating under reduced pressure, drying, Apium graveolens Linnaeus extract content accounts for 10 ~ 100% of active constituents of medicine quality.
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