CN103479711B - Application of eucommia ulmoides lignans extracts in preparation of medicine treating renal interstitial fibrosis - Google Patents
Application of eucommia ulmoides lignans extracts in preparation of medicine treating renal interstitial fibrosis Download PDFInfo
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Abstract
The invention relates to the field of purposes of extracts of traditional Chinese medicine, in particular to application of eucommia ulmoides lignans extracts in preparation of medicine treating renal interstitial fibrosis. Lignans accounts for 30-90% of the total weight of the eucommia ulmoides lignans extracts, and the effective constituent of pinoresinol diglucoside in the lignans accounts for 1%-15% of the total weight of the eucommia ulmoides lignans extracts.
Description
Technical field
The invention belongs to Chinese medicine extract purposes field, be specifically related to the novelty teabag of Cortex Eucommiae lignans extract.
Background technology
Renal fibrosis is a kind of pathophysiological change, be kidney function by health to damage, then to damage, until the progressive process of afunction.The consequently irreversible infringement of renal function Progressive symmetric erythrokeratodermia, brings huge threat to human health.Kidney region fibrosis is the common pathological characters that kidney disease caused by a variety of causes advances to End-stage renal failure.A large amount of clinical and pathological data shows, the order of severity of the Chronic Renal Injury that a variety of causes causes and kidney region fibrosis comparatively glomerulopathy changes to closely.Therefore, in order to slow down and reverse the process of chronic renal disease, improve renal function, find and find the active drug for the treatment of renal fibrosis for guiding clinical treatment, to improve nephrotic's quality of life significant.
The Cortex Eucommiae is the dry bark of the Eucommiaceae plant Cortex Eucommiae, is Chinese famous and precious tonic herb.Tool invigorating the liver and kidney, bone and muscle strengthening, blood pressure lowering, many effects such as antiabortive, it is Chinese peculiar medical material, and its medicinal history is long, has a wide range of applications clinical.The Cortex Eucommiae has invigorating the liver and kidney, bone and muscle strengthening, rubbish in purged body, strengthen human body cell substance metabolism, prevent muscle skeleton aging, balanced human's blood pressure, decomposer inner cholesterol, reduce body fat, recover blood vessel elasticity, diuresis heat clearing away, broad-spectrum antiseptic, stimulating central nervous system, improves white blood cell count, strengthens the remarkable efficacy such as body immunity.The effective ingredient of the Cortex Eucommiae mainly contains lignanoid, iridoids, flavonoid, organic acid, alcohol, trace element, vitamin etc., and lignanoid is one of composition that in the Cortex Eucommiae, content is maximum.Cortex Eucommiae lignanoid has the pharmacological actions such as blood pressure lowering, anti peroxidation of lipid, protection central nervous system.
The domestic and international pharmacological research to Cortex Eucommiae lignans extract is less at present.Application number: 200710035744.7 disclose Cortex Eucommiae lignanoid and the application of extract on anti-cardiovascular reconstruction thereof, the novelty teabag of Cortex Eucommiae lignans extract needs to be studied further.
Summary of the invention
Object of the present invention aims to provide a kind of novelty teabag of Cortex Eucommiae lignans extract.We study and find that Cortex Eucommiae lignans extract has preventive and therapeutic effect to kidney region fibrosis, can be used for the medicine preparing treatment kidney region fibrosis.
For achieving the above object, technical scheme of the present invention is:
The application of Cortex Eucommiae lignans extract in preparation treatment kidney region fibrosis medicine, in described Cortex Eucommiae lignans extract, lignanoid accounts for the 30%-90% of Cortex Eucommiae lignans extract gross weight, and in lignanoid, the content of effective ingredient pinoresinol diglucoside accounts for the 1%-15% of Cortex Eucommiae lignans extract gross weight.
In described lignanoid, the content of effective ingredient pinoresinol diglucoside preferably accounts for the 3%-7.5% of Cortex Eucommiae lignans extract gross weight.
It is raw material that described Cortex Eucommiae lignans extract is preferably with the Cortex Eucommiae, be the alcohol extraction of 40%-95% by mass concentration, extracting liquid filtering, concentrated, concentrated solution is passed through macroporous resin column, use water successively, mass concentration is methanol or the ethanol of 15%-25%, and mass concentration is methanol or the alcoholic solution eluting of 45%-55%, collecting mass concentration is the alcohol eluen of 45%-55%, concentrated, dry, obtain Cortex Eucommiae lignans extract powder.
The preferred preparation method of described Cortex Eucommiae lignans extract is:
A), after Cortex Eucommiae removes crust, cutting is alcoholic solution reflux, extract, 2-3 time of 40%-95% by mass concentration, each 1-2h, and extracting solution is evaporated to 0.5-1.5 times of medical material amount volume;
B) medicinal liquid after concentrated is carried out effective ingredient enrichment by macroporous adsorptive resins, use water successively, mass concentration is methanol or the ethanol of 15%-25%, and mass concentration is methanol or the alcoholic solution eluting of 45%-55%;
C) collecting mass concentration is methanol or the ethanol elution of 45%-55%, concentrated, dry Cortex Eucommiae lignans extract powder.
Collect the water elution liquid in step b, concentrated, dry, pulverize, powder dissolve with methanol, discards insoluble matter, reclaims methanol and dry iridoids material.
Macroporous adsorptive resins described in step b is HP series plastics, D101 resin or AB-8 resin.
Cortex Eucommiae lignans extract of the present invention also can be Cortex Eucommiae lignans extract disclosed in number of patent application 200710035744.7.
Below the present invention be further explained and illustrate.
Described Cortex Eucommiae lignans extract can be selected from pulverizing by employing, squeezes, calcines, grinds, sieves, percolation, extraction, water extraction, alcohol extraction, water precipitating, precipitate with ethanol, ester are carried, the method such as ketone is carried, chromatography, filtration obtains.With this extract for active substance can be made into pharmaceutical preparation, described extract can be the material of extractum form, can be dry extract also can be fluid extract, can be powdered substance, need to make different states according to the difference of preparation.
The present invention also proposes the pharmaceutical preparation of Cortex Eucommiae lignans extract through the applicable any unit dosage form taken of processing preparation, these pharmaceutical preparatioies are selected from following dosage form: tablet, sugar coated tablet, film coated tablet, enteric coated tablet, capsule, hard capsule, soft capsule, oral liquid, suck agent, granule, electuary, pill, powder, unguentum, sublimed preparation, suspensoid, powder, solution, suppository, ointment, plaster, cream, spray, drop, patch, can make slow releasing preparation, enteric coated preparation when needing.
Cortex Eucommiae lignans extract of the present invention, medicine acceptable carrier can be added when being prepared into pharmaceutical preparation, described medicine acceptable carrier is selected from: antioxidant, intercalating agent surfactant filler disintegrating agent wetting agent dispersant lubricant solvent slow release material enteric material pH adjusting agent, correctives, pigment etc., conventional carrier is as mannitol, dextran, lactose, glucose, sorbitol, mannitol, xylitol, sodium chloride, silicon derivative, cellulose and cellulose derivative, sodium sulfite, sodium pyrosulfite, alginate, gelatin, polyvinylpyrrolidone, glycerol, Tween 80, agar, calcium carbonate, calcium bicarbonate, Polyethylene Glycol, cyclodextrin, beta-schardinger dextrin-, phospholipid material, Kaolin, Pulvis Talci, calcium stearate, magnesium stearate etc.
The pharmaceutical preparation of Cortex Eucommiae lignans extract of the present invention is by being processed through extraction or other modes by Cortex Eucommiae raw material, making pharmaceutically active substance, subsequently, with this active substance for raw material, when needing, add medicine acceptable carrier, make pharmaceutical preparation according to the routine techniques of galenic pharmacy.
Accompanying drawing explanation
Fig. 1 is the common light microscopy checking figure (numbering: 1M12) after rats in sham-operated group kidney HE dyes; HE dyes, and × 200; Glomerule.Renal tubules form is normal, the changes such as interstitial has no the congestion of blood vessel, inflammatory cell infiltration.
Fig. 2 is the common light microscopy checking figure (numbering: 2M11) after model group rats kidney HE dyes.HE dyes, and × 200.Glomerule.Renal tubules structure disappears, a large amount of proliferation of fibrous tissue of interstitial, and with a large amount of inflammatory cell infiltration.
Common light microscopy checking figure (numbering: 3M06) after Tu3Shi Cortex Eucommiae lignanoid low dose group rat kidney HE dyes.HE dyes, and × 200.Renal cells degeneration necrosis, part tubule dilatation, a large amount of proliferation of fibrous tissue of interstitial, and with a large amount of inflammatory cell infiltration.
Common light microscopy checking figure (numbering: 4M02) in Tu4Shi Cortex Eucommiae lignanoid after dosage group rat kidney HE dyeing.HE dyes, and × 200.Renal cells degeneration necrosis, part tubule dilatation, a small amount of proliferation of fibrous tissue of interstitial, and with a small amount of inflammatory cell infiltration.
Common light microscopy checking figure (numbering: 5M05) after Tu5Shi Cortex Eucommiae lignanoid high dose group rat kidney HE dyes.HE dyes, and × 200.Renal cells degeneration necrosis, part tubule dilatation, a small amount of proliferation of fibrous tissue of interstitial, and with a small amount of inflammatory cell infiltration.
Fig. 6 is the common light microscopy checking figure (numbering: 6M02) after Fluorofenidone positive controls rat kidney HE dyes.HE dyes, and × 200.Renal cells degeneration necrosis, a small amount of proliferation of fibrous tissue of interstitial, and with a small amount of inflammatory cell infiltration.
Detailed description of the invention
Below by embodiment, the present invention will be further explained, and described in embodiment, percentage composition is mass percentage.
The preparation of embodiment 1 Cortex Eucommiae lignans extract
Get eucommia bark (removing crust) 2Kg, with 8L65% alcohol reflux twice, each 1h, filters, and merges extracted twice liquid and is concentrated into 800ml.Mix sample loading (macroporous resin column volume 3L) with D101 macroporous resin, wash 3 times of column volumes, 1% ammonia continues to wash 3 times of column volumes, then is washed to neutrality.With 20% ethanol elution, 2 times of column volumes, wash 2.5 times of column volumes with 45% ethanol.45% ethanol elution is concentrated into 600ml.Powder is dried to after eluent is concentrated.With pinoresinol diglucoside in contrast, Cortex Eucommiae lignans extract is 85% through ultraviolet detection lignans content, and detecting pinoresinol diglucoside content through HPLC is 12.5%.
The preparation of embodiment 2 Cortex Eucommiae lignans extract
Cortex Eucommiae (removing crust) adds 60% ethanol 9L reflux, extract, 1h after pulverizing, and filters, and filtering residue adds 60% ethanol 7L again and extracts 1h, filters, and merges extracted twice liquid, is concentrated into without alcohol taste, obtains concentrated solution and be about 3L.Concentrated solution, through AB-8 macroporous adsorptive resins, first uses 5L water elution, then uses 7L60% ethanol elution, collects 60% ethanol elution, is concentrated into 150ml, is slowly added in 450ml water, filters, concentrated, obtains drying solid powder.With pinoresinol diglucoside in contrast, Cortex Eucommiae lignans extract is 51% through ultraviolet detection lignans content, and detecting pinoresinol diglucoside content through HPLC is 3.5%.
Embodiment 3
The Protection to the kidney region fibrosis Cortex Eucommiae lignans extract that embodiment 1 and embodiment 2 obtain being used for embodiment 3 is studied.
1. experimental technique
1.1 dose design and grouping
Experiment is divided into sham operated rats, model group, Cortex Eucommiae lignans extract low dose group (150mg/kg), middle dosage group (300mg/kg), high dose group (600mg/kg) and Fluorofenidone positive controls (500mg/kg).
Table 1 tests grouping and dose design
1.2 experimental procedure
Male SD rat 60, body weight 250 ~ 300g, surgical ligation group adopts unilateral ostruction, method is as follows: by 2mL/kg lumbar injection 2% pentobarbital sodium anesthetized rat, aseptic condition is about 1.5cm in abdominal part left kidney position otch, kidney is extruded gently, left side ureter surrounding tissue is peeled off gently with ophthalmic tweezers, expose ureter, with sterilizing stitching thread ligation ureter, separately get 12 rats as sham operated rats, modus operandi is identical, kidney is extruded gently rear separation left side ureter but not ligation.After above animal surgery completes, suture muscles, skin successively, wound surrounding injection 100,000 U penicillin is in case infect.Eye socket blood sampling was carried out to 60 ligation group rats in the 3rd day after surgery, separation of serum, detect serum creatinine and urea nitrogen content, result display surgical ligation group serum creatinine and urea nitrogen content raise, according to index result, surgical ligation group is divided into 5 groups at random, often organize 12, be respectively model group, the basic, normal, high dosage group of Cortex Eucommiae lignans extract and Fluorofenidone positive controls.Cortex Eucommiae lignans extract distilled water, Fluorofenidone 0.5%CMC-Na are according to dosage mixed with respective concentration before experiment by every day, now with the current, by 10mL/kg gastric infusion, and 1 times/day, continuous 15 days.Sham operated rats and model group give isopyknic distilled water.Press 2mL/kg lumbar injection 2% pentobarbital sodium anesthetized rat after last administration and after euthanasia, solution takes nephridial tissue and fixes, paraffin embedding, carry out tissue pathology checking.
1.3 Testing index
Kidney morphologic observation: get rat kidney respectively and be placed in 10% neutral formalin liquid and fix, conventional dehydration, embedding, section, HE dyeing, carry out histopathologic examination.Pathology ranking criterion: tubulointerstitial injury is marked, each specimen Stochastic choice does not repeat 10 visuals field, comprises cortex, medullary substance, skin marrow intersection, comprises 6 specific targets: (1) tubular ectasia; (2) renal tubules atrophy; (3) renal cells cavity is formed; (4) renal cells cavity is formed; (5) kidney region fibrosis; (6) renal interstitial inflammatory cell infiltration, namely the focal type cell infiltration of renal interstitial is mild damage; The inflammatory cell infiltration that diffusivity is dispersed in is moderate infringement; The intensive inflammatory cell infiltration of diffusivity is severe infringement.According to the degree of pathological lesion, 0 point is divided into each project: normal; 1 point: mild damage's (pathological changes is less than 20%); 2 points: moderate infringement (pathological changes 20% ~ 40%); 3 points: severe infringement (pathological changes is greater than 40%).
1.4 statistical method:
Adopt SPSS16.0 to carry out statistical analysis, the level set of statistical significance is P<0.05.Adopt mean ± standard deviation
by Leven ' s test method inspection normality and homogeneity of variance.If meet normality and homogeneity of variance, carry out statistical analysis with one factor analysis of variance (One-way ANOVA) and post Hoc LSD; If do not meet normality and heterogeneity of variance, then check with Kruskal-Wallis.If Kruskal-Wallis inspection has statistical significance (P<0.05), then use Dunnett ' s Test(nonparametric technique) compare analysis.Significant difference and biological significance is considered during evaluation.
2 experimental results
Cortex Eucommiae lignans extract is on the impact of Renal Paphology
As shown in table 2 and Fig. 1-6, comparing with normal control, there is tubular ectasia, atrophy in model group rats kidney, and renal cells is downright bad, kidney region fibrosis and the intensive inflammatory cell infiltration of diffusivity.As can be seen from the statistical result of average pathology integration, model group pathology integration is significantly higher than sham operated rats, prompting renal fibrosis model modeling success.Administration is after 15 days, and Cortex Eucommiae lignans extract high dose group and Fluorofenidone positive controls all occur significant difference compared with model group, and there is a change for the better for kidney region fibrosis.
Table 2 Cortex Eucommiae lignans extract is on the pathological impact of kidney region fibrosis
Note: compare with sham operated rats,
++p<0.01, compares with model group,
*p<0.05.
3 experiment brief summaries
The above results shows, Cortex Eucommiae lignans extract high dose group significantly can improve tubular ectasia, the atrophy of kidney region fibrosis model, and the symptoms such as renal interstitial inflammatory cell infiltration, have some improvement to kidney region fibrosis.
Claims (6)
1. the application of Cortex Eucommiae lignans extract in preparation treatment kidney region fibrosis medicine, in described Cortex Eucommiae lignans extract, lignanoid accounts for the 30%-90% of Cortex Eucommiae lignans extract gross weight, and in lignanoid, the content of effective ingredient pinoresinol diglucoside accounts for the 1%-15% of Cortex Eucommiae lignans extract gross weight.
2. apply according to claim 1, it is characterized in that, in described lignanoid, the content of effective ingredient pinoresinol diglucoside accounts for the 3%-7.5% of Cortex Eucommiae lignans extract gross weight.
3. apply according to claim 1 or 2, it is characterized in that, described Cortex Eucommiae lignans extract take the Cortex Eucommiae as raw material, be the alcohol extraction of 40%-95% by mass concentration, extracting liquid filtering, concentrated, concentrated solution is passed through macroporous resin column, use water successively, mass concentration is methanol or the ethanol of 15%-25%, and mass concentration is methanol or the alcoholic solution eluting of 45%-55%, collecting mass concentration is the alcohol eluen of 45%-55%, concentrated, dry, obtain Cortex Eucommiae lignans extract powder.
4. apply according to claim 3, it is characterized in that, the preparation method of described Cortex Eucommiae lignans extract is:
A), after Cortex Eucommiae removes crust, cutting is alcoholic solution reflux, extract, 2-3 time of 40%-95% by mass concentration, each 1-2h, and extracting solution is evaporated to 0.5-1.5 times of medical material amount volume;
B) medicinal liquid after concentrated is carried out effective ingredient enrichment by macroporous adsorptive resins, use water successively, mass concentration is methanol or the ethanol of 15%-25%, and mass concentration is methanol or the alcoholic solution eluting of 45%-55%;
C) collecting mass concentration is methanol or the ethanol elution of 45%-55%, concentrated, dry Cortex Eucommiae lignans extract powder.
5. apply according to claim 4, it is characterized in that, collect the water elution liquid in step b, concentrated, dry, pulverize, powder dissolve with methanol, discards insoluble matter, reclaims methanol and dry iridoids material.
6. apply according to claim 4, it is characterized in that, macroporous adsorptive resins described in step b is HP series plastics, D101 resin or AB-8 resin.
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| CN110538189A (en) * | 2019-09-22 | 2019-12-06 | 江西普正制药股份有限公司 | Eucommia ulmoides extract composition for treating renal fibrosis and application thereof |
| CN111202741B (en) * | 2020-01-21 | 2021-07-20 | 中南大学湘雅医院 | Application of lignan compound in preparation of medicine for treating and/or preventing nephropathy |
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| CN102643610A (en) * | 2011-02-16 | 2012-08-22 | 严晓敏 | Production method for extracting natural polymer material from eucommia pericarp |
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| CN102643610A (en) * | 2011-02-16 | 2012-08-22 | 严晓敏 | Production method for extracting natural polymer material from eucommia pericarp |
Non-Patent Citations (2)
| Title |
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| 《内蒙古中医药》;韩宇东;《杜仲药理活性研究进展》;20110228(第4期);125-126 * |
| 《杜仲对单侧输尿管梗阻大鼠肾间质纤维化的影响》;骆小娟等;《山东医药》;20121231;第52卷(第28期);47-48 * |
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