A kind of preparation method and applications of father-in-law's cloth extract
Technical field
The present invention relates to a kind of preparation method of father-in-law's cloth extract, the father-in-law's cloth extract prepared with this method is also related to
Application, belong to Tibetan medicine field.
Background technique
Hyperuricemia (hyperuricemia) is one kind due to caused by purine metabolic disturbance and/or uric acid excretion disorder
Blood uric acid increases, and extracellular fluid lithate is in the disease of over-saturation.Blood uric acid can clarify a diagnosis when being more than 416 μm of ol/L as height
Uricacidemia.Superelevation protein meal is the major reason for causing purine metabolic disturbance, excessive glucocorticoid, fat, protein intake and
Heavy drinking and illness rate increase have important relationship.When serum Uric Acid Concentration is excessively high, uric acid is deposited on pass in the form of sodium salt
Section and subcutaneous tissue, lead to the generation of gout, therefore about 5%~12% can develop into gout in Patients with Hyperuricemia.Bitterly
Wind can not only invade bone and joint, but also be also easy to involve kidney and cardiovascular system.Hyperuricemia and gout are current
Seriously endanger the metabolic disease of human health.
Hyperuricemia can be caused by both sides reason:(1) uric acid generation increases:Uric acid is by endogenous and outer in human body
Property two kinds of approach in source generate, and exogenous uric acid derives from food, account for about the 20% of Uric Acid total amount;Endogenous uric acid mainly comes
Derived from nucleotide catabolism, account for about 80%;Therefore, a large amount of decomposition of high purine diet and nucleic acid in vivo can lead to blood uric acid
Raising.Wherein, xanthine oxidase is the key enzyme that uric acid generates, it, which can be catalyzed hypoxanthine and is oxidized to xanthine, reoxidizes
For uric acid.(2) underexcretion:Uric acid mainly passes through kidney excretion, kidney to the filtration for being discharged through glomerulus of uric acid,
Reabsorption is adjusted after the active secretion and secretion of reabsorption, renal tubule before secretion, wherein with renal tubule to uric acid reabsorption
Based on.Lithate is polar molecule, needs ion channel could be by renal cells film, presently found ion channel
In (lithate transport protein), anion exchanger is crucial ion channel, which can promote renal tubule to the weight of uric acid
Absorb, determine uric acid be discharged from kidney number.
Western medicine hyperuricemia is mainly in terms of following two at present:First is that the drug for inhibiting uric acid to generate, such as not fast
Purine alcohol, colchicin, non-steroid anti-inflammatory drug (NSAIDs), glucocorticoid etc.;Second is that promote the drug of uric acid excretion, such as third
Semi-annular jade pendant relaxes, sulfinpyrazone, Benzbromarone etc.;But said medicine has a different degrees of side effect, such as allergic rash, drug fever,
Upset,gastro-intestinal, leucocyte and decrease of platelet, hepatic disorder etc..
Hyperuricemia is physico-chemical examination discovery, and Chinese medicine is without corresponding disease name, but Chinese medicine is to caused by hyperuricemia
Gout is early recognized.《Synopsis Golden Chamber apoplexy joint-running》?:" pulse at CUN KOU being deep and weak, deep pulse indicating the disease affecting bone, weak i.e. main reinforcement, deep pulse indicating the kidney disease,
Weak is liver ... joint-running Huang sweating, therefore day joint-running ".From the point of view of symptom, the gout of modern medicine should belong to the bi Zheng of Chinese medicine,
The scope of " joint-running ".With regard to its etiology and pathogenesis, ancient Chinese medicine doctor discussion is a lot of, and the formation of hyperuricemia is mainly due to native endowment
Deficiency, essence in kidney are in the red, and throw off restraint thermophilic fat meat and fine grain delicious food, the with the passing of time impairment of the spleen, the dysfunction of the spleen in transport day after tomorrow, and kidney loses transpiration, whole body is caused to gasify
Mistake department, body fluid dysbolism eventually lead to interior wet growth, and damp evil disperse is formed in ying blood.Yuan Dynasty's ZHU Dan-xi exists《Further Discourses on the Properties of Things
Gout opinion》Middle theory:" that gout person, big rate is because blood is heated, own self-boiling, thereafter or relates to cold water or vertical wetland, or fan take it is cool, or
Sleeping work as wind, cold and cool to fight outside, warm blood must tremble with fear, sweat is turbid coagulate it is puckery, so having a pain.Night, then very, row was in yin for pain." it is discussed and is set to name of disease
Gout, pathogenic factor are blood-head and wind, are trembled with fear, and wetting phase is fought, and clinic is using pain as main feature.Traditional Chinese medicine treats antihyperuricemic
The research of disease is gradually taken seriously and pays close attention to, and achieves certain achievement, and curative effect has also shown many superior
Property.
Tibetan medicine father-in-law cloth original plant is Chinese tamarisk section (Tamaricaceae) Myricaria German tamarisk Myricaria germanica
(L.) Desv., stem is upright and multi-branched, and branch rufous, leaf is small, linear or linear oblong, be distributed in Tibet, Qinghai, Sichuan,
Yunnan, Gansu, Shaanxi, Shanxi are grown on the river shoal environment of 2000~2900m of height above sea level.According to《Jingzhubencao》It records, father-in-law is furnished with clearly
The effect of thermal detoxification, diverging promoting eruption, Yellow River harnessing edema is the common drug of the minority areas such as China Tibet.But related father-in-law's cloth
Basic research it is still extremely limited, be restricted the subsequent promotion and application of the medicinal material.Chemistry of the modern study about father-in-law's cloth
Ingredient and pharmacological action are rarely reported, and are also had no the preparation method of father-in-law's cloth extract and its are used to prepare treatment hyperuricemia medicine
The report of object.
Summary of the invention
The object of the present invention is to provide a kind of preparation method and applications of father-in-law's cloth extract, can be high effective for treatment
Uricacidemia.
To achieve the goals above, the technical scheme adopted by the invention is that providing a kind of preparation side of father-in-law's cloth extract
Method includes the following steps:
(1) father-in-law's cloth pulverizing medicinal materials cross 20-80 mesh, then carry out supercritical carbon dioxide extracting, extraction temperature 40- to it
50 DEG C, extracting pressure 20-30MPa, extraction time 1-2h, extraction and separation obtain volatile oil and dregs of a decoction A, collect volatile oil, obtain
Father-in-law's cloth extract A;
(2) concentration 60-95% ethyl alcohol is added in the dregs of a decoction A of step (1), 50-80 DEG C of temperature extraction takes 2-4 times, mention every time
Taking the time is 1-3h, and it is 8-12 times of dregs of a decoction A weight that concentration 60-95% ethanol consumption is added every time, filtering, obtain dregs of a decoction B with
Extracting solution, combined extract recycle ethyl alcohol, are concentrated and dried, obtain father-in-law's cloth extract B;
(3) dregs of a decoction B of step (2) is added into water refluxing extraction 1-4 times, each extraction time is 1-3h, and each water consumption is
6-10 times of dregs of a decoction B weight, filtering, combined extract are concentrated into relative density 1.07, are splined on macroporous absorbent resin, first use
Water elution, then with the ethanol elution of concentration 30-70%, ethanol eluate is collected, ethyl alcohol is recycled, is concentrated and dried, obtains Weng Buti
Take object C;
(4) father-in-law's cloth extract A, father-in-law's cloth extract B and Weng Bu extract C are uniformly mixed, obtain father-in-law's cloth extract.
The macroporous absorbent resin of step (3) is HPD600 macroporous absorbent resin or LSA-21 macroporous absorbent resin.
The technical solution adopted in the present invention, which also resides in, to be provided father-in-law's cloth extract of above method preparation a kind of and controls in preparation
Treat the application in antihyperuricemic disease drug.
Father-in-law's cloth extract prepared by the present invention can be made into tablet, capsule or granule, and method therefor is this field
Conventional technical means.
The present invention is for the first time with Chinese tamarisk section Myricaria German tamarisk Myricaria germanica (L.) Desv. (father-in-law's cloth), system
Standby father-in-law's cloth extract, raw material sources are abundant, and at low cost, easy to use, quick, patient suffering is small, not anti-medicine, toxic side effect
Small, safety coefficient is high.The preparation method of father-in-law's cloth extract of the present invention is simple, easy to operate, and prepared father-in-law's cloth extract can
Effective for the treatment of hyperuricemia, the research promotion of father-in-law's cloth has effectively been pushed.Experiment shows that father-in-law's cloth extract can be significant
Reduce Patients with Hyperuricemia blood uric acid, creatinine, urea nitrogen content and hyperuricemia model group rat blood uric acid, urine uric acid,
Xanthine oxidase, serum creatinine, urea nitrogen content.
Specific embodiment
Specific embodiments of the present invention will be described in further detail with reference to embodiments.
Embodiment 1
The preparation method of father-in-law's cloth extract of the present embodiment, includes the following steps:
(1) father-in-law's cloth pulverizing medicinal materials cross 20 meshes, then carry out supercritical carbon dioxide extracting to it, 45 DEG C of extraction temperature, extraction
Pressure power 20MPa, extraction time 1h, extraction and separation obtain volatile oil and dregs of a decoction A, collect volatile oil, obtain father-in-law's cloth extract A;
(2) 60% ethyl alcohol of concentration is added in the dregs of a decoction A of step (1), 50 DEG C of temperature extractions take 2 times, and each extraction time is
10 times that 60% ethanol consumption of concentration is dregs of a decoction A weight are added in 1h every time, and filtering obtains dregs of a decoction B and extracting solution, merge and extract
Liquid recycles ethyl alcohol, is concentrated and dried, obtains father-in-law's cloth extract B;
(3) dregs of a decoction B of step (2) is added water refluxing extraction 1 time, extraction time 1h, water consumption is the 6 of dregs of a decoction B weight
Times, extracting solution is concentrated into relative density 1.07, is splined on HPD600 macroporous absorbent resin, is first eluted with water by filtering, then with dense
The ethanol elution of degree 30% collects ethanol eluate, recycles ethyl alcohol, is concentrated and dried, obtains father-in-law's cloth extract C;
(4) father-in-law's cloth extract A, father-in-law's cloth extract B and Weng Bu extract C are uniformly mixed, obtain father-in-law's cloth extract.
Embodiment 2
The preparation method of father-in-law's cloth extract of the present embodiment, includes the following steps:
(1) father-in-law's cloth pulverizing medicinal materials cross 80 meshes, then carry out supercritical carbon dioxide extracting to it, 50 DEG C of extraction temperature, extraction
Pressure power 30MPa, extraction time 2h, extraction and separation obtain volatile oil and dregs of a decoction A, collect volatile oil, obtain father-in-law's cloth extract A;
(2) 95% ethyl alcohol of concentration is added in the dregs of a decoction A of step (1), 80 DEG C of temperature extractions take 4 times, and each extraction time is
8 times that 95% ethanol consumption of concentration is dregs of a decoction A weight are added in 3h every time, and filtering obtains dregs of a decoction B and extracting solution, merge and extract
Liquid recycles ethyl alcohol, is concentrated and dried, obtains father-in-law's cloth extract B;
(3) dregs of a decoction B of step (2) is added water refluxing extraction 4 times, each extraction time is 3h, and each water consumption is dregs of a decoction B
10 times of weight, filtering, combined extract are concentrated into relative density 1.07, are splined on HPD600 macroporous absorbent resin, first use water
Elution, then use the ethanol elution of concentration 70%, collect ethanol eluate, recycle ethyl alcohol, concentrate drying, obtain father-in-law's cloth extract C;
(4) father-in-law's cloth extract A, father-in-law's cloth extract B and Weng Bu extract C are uniformly mixed, obtain father-in-law's cloth extract.
Embodiment 3
The preparation method of father-in-law's cloth extract of the present embodiment, includes the following steps:
(1) father-in-law's cloth pulverizing medicinal materials cross 40 meshes, then carry out supercritical carbon dioxide extracting to it, 40 DEG C of extraction temperature, extraction
Pressure power 25MPa, extraction time 1.5h, extraction and separation obtain volatile oil and dregs of a decoction A, collect volatile oil, obtain father-in-law's cloth extract
A;
(2) 75% ethyl alcohol of concentration is added in the dregs of a decoction A of step (1), 70 DEG C of temperature extractions take 3 times, and each extraction time is
12 times that 75% ethanol consumption of concentration is dregs of a decoction A weight are added in 2h every time, and filtering obtains dregs of a decoction B and extracting solution, merge and extract
Liquid recycles ethyl alcohol, is concentrated and dried, obtains father-in-law's cloth extract B;
(3) dregs of a decoction B of step (2) is added water refluxing extraction 2 times, each extraction time is 2h, and each water consumption is dregs of a decoction B
8 times of weight, filtering, combined extract are concentrated into relative density 1.07, are splined on LSA-21 macroporous absorbent resin, first use water
Elution, then use the ethanol elution of concentration 50%, collect ethanol eluate, recycle ethyl alcohol, concentrate drying, obtain father-in-law's cloth extract C;
(4) father-in-law's cloth extract A, father-in-law's cloth extract B and Weng Bu extract C are uniformly mixed, obtain father-in-law's cloth extract.
The clinical observation on the therapeutic effect of 1 father-in-law's cloth extract for treating hyperuricemia of experimental example
Father-in-law's cloth extract of the present invention is in May, 2013 to clinical observation in December, 2014 Patients with Hyperuricemia 64, report
It accuses as follows:
1, general information
In 64 patients, male 34, women 30, the age is 35 years old minimum, 64 years old maximum, and average age 50.7 years old, disease
Journey 6 months to 10 years.
2, diagnostic criteria
Reference《New Chinese medicine guideline of clinical investigations》:
(1) diagnostic criteria of hyperuricemia is:Blood uric acid (UA) >=416 μm ol/L;
3, it is included in and exclusion criteria
It is included in standard:1. meeting diagnostic criteria person;2. age 20-70 year person;3. not appointing in 2 months before all morbidities
What is treated;4. signing informed consent form, agreement receives treatment method and obeys arranger.Exclusion criteria:1. not meeting above-mentioned be included in
Standard person;2. concurrent infection disease, immunity disease, cancer, serious liver and kidney disease person;3. applying cortex class in 12 months
Steroid hormone or immunosuppressor person.
4, therapeutic scheme
Give 64 Patients with Hyperuricemia for meeting above-mentioned standard to drug:Father-in-law's cloth extract system prepared by embodiment 3
At capsule, 4 tablets each time, 2 times a day, it even served 4 weeks.
5, treatment results
After medication 4 weeks, blood uric acid, serum creatinine, urea nitrogen detection are carried out, curative effect is evaluated.
(1) detection data compares
Blood uric acid, serum creatinine, urea nitrogen before and after measurement patient takes medicine respectively, see Table 1 for details:
The pretherapy and post-treatment blood uric acid of 1 patient of table, serum creatinine, urea nitrogen variation
Note:Compared with before treatment,*P<0.05
(2) therapeutic evaluation:
Curative effect determinate standard:It is effective:Blood uric acid reduces >=15%;Effectively:Blood uric acid reduces<15%;In vain:UA level with
Compared to unchanged or raising before treatment.Total effective rate=obvious effective rate+effective percentage.
Therapeutic evaluation:It is effective 41 (64.1%) in 64 patients, it is effective 20 (31.2%), it is invalid 3 (4.7%),
Total effective rate is 95.3%.
Influence of the 2 father-in-law's cloth extract of experimental example to hyperuricemia rat
1, experimental animal
SD rat, 60, male, is purchased from experimental animal center of henan province, the quality certification by SPF grades, weight 200g~220g
Number:SCXK (Henan) 2010-0002.
2, drug and instrument
Father-in-law's cloth extract particles agent of the present invention (embodiment 2 is prepared);Allopurinol tablet (upper Hisense's friendship everything medicine company share
Co., Ltd, lot number 20130116);Oxonic Acid sylvite (Sigma company, lot number STBC6418V);(Amresco is public for adenine
Department, lot number 130410), uric acid (lot number 20130519), creatinine (lot number 20130424), urea nitrogen (lot number 20130509), Huang
Purine oxidase (lot number 20130521) kit is purchased from Nanjing and builds up Bioengineering Research Institute;Synergy NEO type enzyme mark
Instrument (Bio-Tek company of the U.S.);2K15 type refrigerated centrifuge (German Sigma company).
3, experimental method
Male SD rat 60,6 groups are randomly divided into, i.e. Normal group, model control group, father-in-law's cloth extract is high, medium and low
Dosage group (father-in-law's cloth extract 120,60,30mg/kg), positive control Allopurinol group (30mg/kg) group.Except for the normal group, each group
Rat oral gavage adenine (250mg/kg, 1 time a day) is simultaneously injected intraperitoneally Oteracil Potassium (200mg/kg, 1 times a week), 4 weeks, structure totally
Build rat high lithemia model.Relative medicine is given in modeling beginning stomach-filling in the 3rd week, 1 time a day, continuous 4 weeks, normal group, model
Same volume distilled water is given in group stomach-filling.
Rat is put into metabolic cage in the 27th day early morning is administered, morning next day collects urinates for 24 hours, remembers urine volume, urine 3000r/min
It is centrifuged 15min, supernatant is taken, urine uric acid is detected by automatic clinical chemistry analyzer.After 28th day fasting 12h, rat abdominal cavity note
10% chloraldurate 0.3mL/kg of mass fraction anesthesia is penetrated, abdominal aorta blood sampling separates serum, measures uric acid in serum (UA),
Urea nitrogen (BUN), serum creatinine (Cr), xanthine oxidase (XOD) content.
4, statistical procedures
It is for statistical analysis to result using 17.0 software of SPSS, data withIt indicates, using single factor test variance point
It analyses, compares two-by-two between group and examined with LSD, P<0.05 is statistically significant (the results are shown in Table 2,3).
5, result
2 father-in-law's cloth extract of table is to hyperuricemia rat blood uric acid, xanthine oxidase and the influence for urinating uric acid
Note:Compared with model control group,*P<0.05
From table 2 it can be seen that the urine uric acid of model group rats significantly reduces, blood uric acid, xanthine oxidase compared with normal group
It is significantly raised to change enzyme, difference has conspicuousness (P<0.05), hints model constructs successfully.Compared with model group, father-in-law's cloth extract is each
Dosage can significantly increase hyperuricemia rat urine uric acid content (P<0.05) rat blood uric acid, xanthine oxidation, are significantly reduced
Enzyme content (P<0.05).
Influence of the 3 father-in-law's cloth extract of table to hyperuricemia rat creatinine, urea nitrogen
Note:Compared with model control group,*P<0.05
From table 3 it can be seen that the content of the serum creatinine of model group rats, urea nitrogen is significantly raised, poor compared with normal group
It is different to have conspicuousness (P<0.05).Compared with model group, the basic, normal, high dosage of father-in-law's cloth extract group can significantly reduce antihyperuricemic
Creatinine, the urea nitrogen content (P of disease rat<0.05).