CN103405494A - Bauhinia championii n-butyl alcohol extract and preparation method and application thereof - Google Patents
Bauhinia championii n-butyl alcohol extract and preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a bauhinia championii n-butyl alcohol extract, which contains beta-sitosterol with the weight percentage not less than 0.1%. The invention also provides a preparation method and application of the bauhinia championii n-butyl alcohol extract. The n-butyl alcohol part of the bauhinia championii is used for treating rheumatoid arthritis, has definite drug effect, controllable quality and stable preparation method, is suitable for large-scale production, and provides a new choice for clinic.
Description
Technical field
The present invention relates to a kind of Caulis Bauhiniae Championii ethyl acetate extract, n-butanol extract and its production and use, belong to drug world.
Background technology
Caulis Bauhiniae Championii is the rattan of pulse family bauhinia Caulis Bauhiniae Championii Bauhinia championii Benth., claims again Caulis Bauhihiae Championii, Flos Mume nuphar japonicum, and mainly distribute Fujian, Jiangxi, Guangxi, Zhejiang etc., have the effect of wind-damp dispelling, promoting the circulation of blood gas.Modern pharmacology studies have shown that, the effective ingredient of Caulis Bauhiniae Championii has the effect that expelling wind and removing dampness, promoting blood circulation and stopping pain, spleen invigorating are regulated the flow of vital energy, and is used for the treatment of rheumatic arthritis, lumbago and skelalgia, traumatic injury, stomachache, infantile malnutrition.According to the south of Fujian Province well-known doctor's gold steel proved recipe, the Caulis Bauhiniae Championii of take has the effect of good treatment burn and scald clinically as the health skin Caring burn cream of main component.The chemical composition of Caulis Bauhiniae Championii medical material is mainly flavonoid, cyanogen glycosides, gallic acid etc.
Document HONG Z F, ZHENG H Y, XU W, et al.Study on resistinflammation and analgesic effect of Kangmeifu BurnsOintment (KBO) [J] .Chin J Tradit Med Sci Technol (Chinese Chinese medicine science and technology), 2007, 14 (4): 261-262.HONG Z F, ZHENG H Y, WANG R G, et al.Study on antibacterial activity of Kangmeifu Burns Ointment (KBO) [J] .Chin J Microecol (Chinese microecology magazine), 2007, 19 (2): 177-178.] reported that flavones ingredient in the Caulis Bauhiniae Championii has the effect of antalgic and inflammation relieving, the document only mentions that thick total flavones has analgesic and anti-inflammatory effects, do not relate to particular compound.Also there is bibliographical information to study chemical composition in Caulis Bauhiniae Championii, as: Bai Haiyun, Deng, Caulis Bauhihiae Championii chemical constitution study (I), CHINA JOURNAL OF CHINESE MATERIA MEDICA, the 30th the 1st phase of volume, in January, 2005, reported from isolation identification ethanol extraction 5 compounds, it extracts and separation method is: get Kowloon rattan 7Kg, pulverize afterwards with 95% industrial alcohol percolation 3 times.Extractum water suspendible, use petroleum ether, ethyl acetate, n-butanol extraction successively.Ethyl acetate extract separates through silica gel, macroporous resin, polydextran gel and reversed-phase silica gel column chromatography repeatedly, gets 5 compounds.
Bai Haiyun etc., Caulis Bauhihiae Championii chemical constitution study (II), Natural products research is developed, 2004, Vol.16NO.4, from ethyl acetate effective site, separating and obtained gallic acid, sitosterol, daucosterol, Quercitroside, myricetrin, epicatechin, 5,6,7,3 ˊ, 4 ˊ, 5 ˊ-hexa methoxy flavone.
Summary of the invention
Technical scheme of the present invention has been to provide a kind of Caulis Bauhiniae Championii ethyl acetate extract, n-butanol extract.Another technical scheme of the present invention has been to provide preparation method and the purposes of this extract.
The invention provides a kind of Caulis Bauhiniae Championii ethyl acetate extract, the weight percentage that it contains Quercetin must not be less than 0.05%, and the weight percentage of ampelopsin must not be less than 0.04%.
Further preferably, the weight percentage that it contains Quercetin is 0.05-1.50%, and the weight percentage of ampelopsin is 0.04-1.25%.
Still more preferably, it contains compounds I, II, III, IV, V.
Caulis Bauhiniae Championii ethyl acetate extract of the present invention is to be prepared from by following steps:
Get the Caulis Bauhiniae Championii medical material, add the 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol to nothing alcohol flavor, and the alcohol extract petroleum ether extraction, obtain petroleum ether layer and water layer, and water layer adds the chloroform extraction, obtains chloroform layer and water layer; The water layer ethyl acetate extraction, obtain acetic acid ethyl acetate extract, reclaims, and dries, and obtains the Caulis Bauhiniae Championii ethyl acetate extract.
Wherein, described ethanol is that concentration is 80% ethanol.
The present invention also provides a kind of method for preparing described Caulis Bauhiniae Championii ethyl acetate extract, comprises the steps:
Get the Caulis Bauhiniae Championii medical material, add the 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol to nothing alcohol flavor, and the alcohol extract petroleum ether extraction, obtain petroleum ether layer and water layer, and water layer adds the chloroform extraction, obtains chloroform layer and water layer; The water layer ethyl acetate extraction, obtain acetic acid ethyl acetate extract, reclaims, and dries, and obtains the Caulis Bauhiniae Championii ethyl acetate extract.
The present invention also provides the purposes of Caulis Bauhiniae Championii ethyl acetate extract in the medicine for preparing antalgic and inflammation relieving, resisting rheumatoid arthritis.
The invention provides a kind of Caulis Bauhiniae Championii n-butanol extract, the weight percentage that it contains cupreol must not be less than 0.1%.
Wherein, its weight percentage of containing cupreol is 0.1-0.85%.
Further, it contains compound V, VI.
Caulis Bauhiniae Championii n-butanol extract of the present invention is to be prepared from by following method:
Get the Caulis Bauhiniae Championii medical material, add the 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol to nothing alcohol flavor, and the alcohol extract petroleum ether extraction, obtain petroleum ether layer and water layer, and water layer adds the chloroform extraction, obtains chloroform layer and water layer; The water layer ethyl acetate extraction, obtain ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, and obtains the Caulis Bauhiniae Championii n-butanol extract.Further preferably, described ethanol is that concentration is 80% ethanol.
The present invention also provides a kind of method for preparing described Caulis Bauhiniae Championii n-butanol extract, and it comprises the steps:
Get the Caulis Bauhiniae Championii medical material, add the 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol to nothing alcohol flavor, and the alcohol extract petroleum ether extraction, obtain petroleum ether layer and water layer, and water layer adds the chloroform extraction, obtains chloroform layer and water layer; The water layer ethyl acetate extraction, obtain ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, and obtains the Caulis Bauhiniae Championii n-butanol extract.
The present invention also provides the purposes of this Caulis Bauhiniae Championii n-butanol extract in the medicine for preparing antalgic and inflammation relieving, resisting rheumatoid arthritis.
Caulis Bauhiniae Championii ethyl acetate extract of the present invention and n-butanol portion are used for the treatment of rheumatoid arthritis, and drug effect is clear and definite, quality controllable, and preparation method is stable, are applicable to large production, provide a kind of new selection for clinical.
The accompanying drawing explanation
Fig. 1 cupreol structural formula figure
Fig. 2 daucosterol structural formula figure
Fig. 3 Quercetin structural formula figure
Fig. 4 compound VI structural formula figure
Fig. 5 rat general state
Fig. 6 Normal group
Fig. 7 model group
Fig. 8 tripterygium glycosides group
Fig. 9 ethyl acetate low dose group
Dosage group in Figure 10 ethyl acetate
Figure 11 ethyl acetate high dose group
Figure 12 n-butyl alcohol low dose group
Dosage group in Figure 13 n-butyl alcohol
Figure 14 n-butyl alcohol high dose group
The specific embodiment
Component separating and the purification of embodiment 1 Caulis Bauhiniae Championii ethyl acetate extract and n-butanol portion
1. extract and separate
Get 1000g Caulis Bauhiniae Championii medical material, add 80% alcohol heating reflux and extract 3 times, each 1.5h, filter, and merges ethanol, reclaims ethanol to without the alcohol flavor, obtains alcohol-extracted extract.Add 2000ml water, make the extractum suspendible, add petroleum ether extraction 5 times, 1000ml/ time, merge petroleum ether extract, reclaim, obtain grease; Water layer adds chloroform extraction 5 times, and 1000ml/ time, the combined chloroform extracting solution, reclaim, and dries, and obtains extractum; Water layer adds ethyl acetate extraction 5 times, and 1000ml/ time, the combined ethyl acetate extracting solution, reclaim, and dries, and obtains extractum; Water layer adds n-butanol extraction 5 times, and 1000ml/ time, merge n-butanol extracting liquid, reclaim, dry, obtain extractum.
The Caulis Bauhiniae Championii ethyl acetate extract that this method obtains calculates by extractum, and yield must not be less than 2%, contains Quercetin and ampelopsin and must not be less than respectively 0.05% and 0.04%.
The Caulis Bauhiniae Championii n-butanol portion that this method obtains calculates by extractum, and yield must not be less than 6%, contains cupreol and must not be less than 0.1% respectively.
Concrete assay method is:
Adopt the HPLC method to measure the content of Quercetin, ampelopsin in the Caulis Bauhiniae Championii ethyl acetate extract.Chromatographic condition is acetonitrile (A)-0.05% phosphoric acid solution (B), the gradient elution program be the 0-5min acetonitrile from 20% to 30%, 5-10min from 30% to 50%, then keep 20min; Flow velocity is 1.0ml/min; The mensuration wavelength is 370nm; Column temperature is 30 ℃.
Triplicate experiment, method is measured the content that extract obtains Quercetin and is respectively 0.062%, 0.064%, 0.065% thus, and the content of ampelopsin is respectively 0.051%, 0.053%, 0.055%, and average content is respectively 0.064%, 0.053%.
Adopt the HPLC method to measure the content of cupreol in the Caulis Bauhiniae Championii n-butanol extract.Chromatographic condition is that acetonitrile (A)-water (B) (96:4); Flow velocity is 1.0ml/min; The mensuration wavelength is 210nm; Column temperature is 30 ℃.
Triplicate experiment, method is measured the content that extract obtains cupreol and is respectively 0.364%, 0.363%, 0.363% thus.Average content is 0.363%.
Ethyl acetate partly uses thick silica gel (160～200 order) 1:1 to mix sample, add with in dichloromethane wet method dress silicagel column, use the methylene chloride-methanol gradient elution, equal-volume is collected eluent, flow point is known and is remerged through the TLC inspection, silica gel, polyamide, SephadexLH-20, ODS separation and purification through repeatedly, obtain compounds I, II, III, IV, V.
N-butyl alcohol is partly used the hot water dissolving, sample is mixed with thick silica gel (160～200 order) 1:1 in the insoluble position of water, add with in dichloromethane wet method dress silicagel column, use the methylene chloride-methanol gradient elution, equal-volume is collected eluent, flow point is known and is remerged through the TLC inspection, and silica gel, polyamide, SephadexLH-20, ODS separation and purification through repeatedly, obtain compound V, VI.
2. Structural Identification
Compounds I: white, needle-shaped crystals (acetone), mp141-142 ℃.5% sulphuric acid-alcoholic solution heating displaing amaranth, vanillin-concentrated sulphuric acid displaing amaranth.In IR, be presented at 3432,2936,2860,1461,1381,1054,959,800cm
-1There is absorption band at place, reaches more than 90% with the matching degree of cupreol in Sigma Biological Sample Library spectrum storehouse.Under petroleum ether-ethyl acetate, petroleum ether-acetone, three kinds of different unfolding conditions of dichloromethane-acetone, its Rf value is consistent with sitosterol, and mixed melting point does not descend.Consistent with document, therefore be accredited as cupreol, structural formula is shown in Fig. 1.
Compound ii: white powder, mp288～289 ℃.In IR, be presented at 3397,2959,2933,2869,1463,1379,1073,1024cm
-1There is absorption band at place, reaches more than 90% with the matching degree of daucosterol in Aldrich Condensed Phase Sample Library spectrum storehouse.On silica gel thin-layer plate, launch by systems such as dichloromethane-acetone, methylene chloride-methanol, methylene chloride-methanol-water, be single-point, its Rf value is consistent with the daucosterol standard substance, and mixed melting point do not descend, therefore be accredited as daucosterol, structural formula is shown in Fig. 2.
Compound III: white plates crystallization (petroleum ether-ethyl acetate), mp65～66 ℃.In IR, be presented at 2917,2848,1472,1061,719cm
-1Absorption band is arranged, with the matching degree of melissane in Aldrich Condensed Phase Sample Library spectrum storehouse, reach more than 90%, and with different dicyandiamide solutions, launch on silica gel thin-layer plate, be single-point, its Rf value is consistent with the melissane standard substance, therefore be accredited as positive melissane.
Compounds Ⅳ: white plates solid (petroleum ether-ethyl acetate), mp97～98 ℃.In IR, be presented at 2917,2849,1742,1472,1363,1239,1179cm
-1There is absorption band at place, with the matching degree of hexacontane in Aldrich Condensed Phase Sample Library spectrum storehouse, reaches more than 90%, and with different dicyandiamide solutions, launches on silica gel thin-layer plate, is single-point, therefore be accredited as positive hexacontane.
Compound V: yellow acicular crystal (chloroform-methanol), mp>300 ℃.Hydrochloric acid-magnesium powder reacting positive.In IR, be presented at 2918,2849,1664,1610,1521,1450,1381,1318,1261,1198,1167,1013cm
-1There is absorption band at place, with the matching degree of Quercetin in HR Aldrich Aldehydes and Ketones spectrum storehouse, reaches more than 90% 1H-NMR data and bibliographical information
[7]The Quercetin data consistent.And on silica gel thin-layer plate, launch with different dicyandiamide solutions, be single-point, and Rf value is consistent with the Quercetin reference substance, mixed melting point does not descend.Therefore be accredited as Quercetin, structural formula is shown in Fig. 3.
Compound VI: yellow particle shape crystal (chloroform-methanol), mp>280 ℃.In IR, being presented at 3409,1647,1611,1576,1458,1293,1257,1168,1119cm-1 has absorption band, through hydrogen spectrum, Mass Spectrometric Identification, infers that the molecular weight of compound is: 326, and preliminary putative structure such as Fig. 4.
Below by concrete pharmacodynamics test proof beneficial effect of the present invention.
Medicine and reagent:
Tripterygium wilfordii Polyglycosidium Tablets, Huangshi Feiyun Pharmaceutical Co., Ltd.'s production (authentication code:, the accurate word Z42021212 of traditional Chinese medicines);
Medicine of the present invention (Caulis Bauhiniae Championii ethyl acetate extract, the n-butanol extract of embodiment 1 preparation); The low dosage of medicine of the present invention is for containing ethyl acetate extract or n-butanol portion 500mg/mL, middle dosage for containing ethyl acetate extract or n-butanol portion 250mg/mL, high dose for containing ethyl acetate extract or n-butanol portion 125mg/mL.
Carrageenin (carrageenan) is provided by Fujian Province medicine inspecting institute, is made into 1% with normal saline, uses after placement 2d.Sodium sulfide: the Shanghai inferior chemical science and technology of system Development Co., Ltd, lot number: 051212.Dimethylbenzene: AR, chemical reagent factory of Shanghai Xingda, lot number: 20050203.Ether: AR, Shanghai City hundred million chemical reagent company limiteies of a specified duration, lot number: 20050401.Cattle II Collagen Type VI, complete Freund's adjuvant, produced by U.S. SIGMA company, purchased from Beijing Bo Leide bio tech ltd.TNF-α, IL-1 (ELISA method), immunoglobulin (1gG, IgM) test kit: purchased from Wuhan Boster Biological Technology Co., Ltd..
The antiinflammatory experiment of test example 1 medicine of the present invention
The impact of 1 xylol induced mice auricle edema: by 80 body weight 20 ± 2g gram mices, by body weight, be divided at random 8 groups: low dosage medicine group of the present invention, middle dosage medicine group of the present invention, high dose medicine group of the present invention, positive controls (Tripterygium wilfordii Polyglycosidium Tablets), model control group (white vaseline).8 groups of mice difference gavages, every day 1 time, continuous 3d, after last coating 30min, with dimethylbenzene, evenly smear in identical place, auris dextra both sides with 0.1ml/ amount only, left ear compares, after 4h, put to death, cut immediately ears, with diameter 8mm card punch, get the same area auricle, on ten thousand/semi-automated analysis balance, weigh, heavily be swelling rate by take (auris dextra weight-left ear weight)/left ear, (the average swelling of model group-average swelling of administration the group)/average swelling of model group of take is inhibitory rate of intumesce.The swelling degree of matched group and administration group is used to SPSS statistical software Treatment Analysis, and between its average, difference adopts one factor analysis of variance, in Table 1.
The impact (n=10) of table 1 medicine group of the present invention on mice caused by dimethylbenzene xylene inflammation
With model control group, compare, each administration group has significant difference (P<0.01, P<0.05), Three doses medicine group of the present invention xylol causes scorching suppression ratio and all is greater than 20%, points out medicine group xylol induced mice auricle edema of the present invention inhibited.And be dose-effect relationship.
2 on Carrageenan cause the impact of rat paw edema: by 80 body weight 150 ± 10g gram SD rats, be divided at random 8 groups by body weight: model group, the basic, normal, high dosage group of medicine of the present invention, Tripterygium wilfordii Polyglycosidium Tablets group.Rat right hind leg is stretching, the self-sustaining sole of the foot subcutaneous injection 1% carrageenin 0.1mL in middle part.Respectively at 0.5h, lh, 1.5h, 2h, 4h, 6h after injecting, use the thickness of vernier caliper measurement swelling limbs, with (cause scorching after-cause scorching front foot sole of the foot thickness)/cause scorching front foot sole of the foot thickness * 100% to calculate swelling rate, with (the average swelling rate of the average swelling rate-experimental group of model control group)/average swelling rate of model control group * 100%, calculate suppression ratio, each is organized date processing and uses SPSS statistical software Treatment Analysis, between its average, difference adopts one factor analysis of variance, the results are shown in Table 2.
Table 2 pharmaceutical composition of the present invention causes the impact (n=10) of pedal swelling suppression ratio on rat carrageenan
Result shows, each medicine group in cause scorching after 1h to 6h with model group, significant difference (P<0.05) is arranged more all, point out rat paw edema due to each medicine group on Carrageenan that inhibitory action is all arranged.Three dosage of medicine group of the present invention, the 1h suppression ratio just reaches more than 30% after causing inflammation; During 1h, peak, its high dose group and Tripterygium wilfordii Polyglycosidium Tablets group comparing difference have the meaning (P<0.O1) of very significant.The antiinflammatory persistent period of low dosage medicine group of the present invention reaches 4h, and more than middle high dose reaches 6h, when 4h and 6h, carry out one factor analysis of variance and show, basic, normal, high dosage and Tripterygium wilfordii Polyglycosidium Tablets no significant difference (P > 0.05).
The analgesic experiment of test example 2 medicines of the present invention (on the impact of female mice hot plate induced pain)
Get the female mice of body weight 20 ± 2g, press hot plate method and before test, measure 80 of the mice of pain threshold in 5s-30s on the hot plate dolorimeter, by body weight, be divided at random 8 groups: blank group, the basic, normal, high dosage group of medicine group of the present invention, Tripterygium wilfordii Polyglycosidium Tablets group.Take and metapedes reaction occurs licking and be observation index.Respectively at the above medicine of every Mus gavage, 0.2mL/ only.0.5h by upper method, measure pain threshold afterwards, then by front method administration, measure the pain threshold of 1h, 1.5h, 2h.Compare the variation of the threshold of pain, administration front and back, and press (after medication-the front average pain threshold of medication)/the front on average pain threshold of medication * 100% calculating threshold of pain raising percentage rate.Each is organized date processing and uses SPSS statistical software Treatment Analysis, and between its average, difference adopts one factor analysis of variance, the results are shown in Table 3.
The impact (n=10) of table 3 medicine on mice hot plate induced pain threshold and threshold of pain increase rate
Result shows, pharmaceutical composition of the present invention and model control group relatively have significant difference (P<0.O5), and still have analgesic effect during 2h.Three dosage of pharmaceutical composition of the present invention all can obviously improve the mice pain threshold.The prompting: pharmaceutical composition of the present invention has remarkable analgesic activity bitterly to thermic, and the analgesia reach 2h lastingly more than.
Test example 3 medicine of the present invention is on arthritic impact
Grouping and modeling: Wistar rat (male), average weight (200 ± 20) g, adaptability raised for 1 week, unified illumination, low temperature environment, freely drink water, and solid feed is freely ingested.Be divided at random 9 groups, be dosage group (H), n-butyl alcohol high dose group (I) in dosage group (E), ethyl acetate high dose group (F), n-butyl alcohol low dose group (G), n-butyl alcohol in normal group (A), model group (B), tripterygium glycosides group (C), ethyl acetate low dose group (D), ethyl acetate, every group is 10.Complete Freund's adjuvant (CFA) is mixed with the cattle II Collagen Type VI emulsion equal-volume of 2mg/ml, make collagen Emulsion, except normal group, every intradermal injection collagen protein 0.2mg of rat root of the tail section, namely started the gastric infusion that divides into groups, every day 1 time on the 1st after initial immunity.In the 7th day, carry out secondary immunity, except normal group, every rat root of the tail section intradermal injection collagen Emulsion 0.1mg(incomplete Freund's adjuvant (IFA) mixes with the cattle II Collagen Type VI emulsion equal-volume of 2mg/ml, makes collagen Emulsion).Normal group, model group give 9.0g/L sodium chloride injection 10ml/kg every day; The high dose group gavage gives 500mg/kg, and middle dosage group gavage every day gives 250mg/kg, and low dose group gavage every day gives 125mg/kg, and the tripterygium glycosides group gives 6mg/kg every day.All animals successive administration 28 days.Adopt volumetric method 1d before modeling to measure rats with bilateral metapedes pawl volume, cause scorching rear every 4d rats with bilateral foot pawl volume is measured, record volumetric values, using this as the radix of estimating antiinflammatory action intensity.
Sample collection and index detect: before molding, after modeling, survey the variation of pedal swelling thickness and body weight respectively; Before treatment, after treatment, calculate the arthritis swelling; After treating administration in the 28th day, the all animals abdominal aortic blood, divide and be put in drying tube, drying tube is placed in to 3000 rev/mins of centrifugal 8min, get the supernatant separation of serum, press respectively the content that test kit description method is measured TNF-α, interleukin-6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10).
Statistical method: adopt SPSS15.0 data statistics bag to carry out date processing, experimental data is used
Mean.Paired t-test and between group variable analysis in measurement data employing group.
1 rat general state is observed as shown in Figure 5, and the model group rat is after collagen emulsion sensitization, booster immunization, and the mental status, diet situation, mobility decrease, and after initial immunity, festering appears in afterbody, incrustation gradually after 7 days.After secondary immunity about the 3rd day, rat starts morbidity, and rat model toes and ankle joint start to occur swelling, and day by day increase the weight of, and the toes distortion appears in weight loss, part Rat Right metapedes, without dead; After positive controls and the treatment of Caulis Bauhiniae Championii group rat, above-mentioned symptom is obviously alleviated.
The body weight of 2 rats: the results are shown in Table 4.Except the medicine group, other are respectively organized rat body weight and slowly increase, at 14 days all during 21 days body weight start to descend, then slowly increase.
The impact of each group of table 4 on rat body weight
Annotate: with normal group, compare,
#P ﹤ 0.05,
##P ﹤ 0.01; With model group, compare,
*P ﹤ 0.05,
*P ﹤ 0.01;
3 pathological study: each treated animal ankle joint pathology demonstration, the synovial cells in rats synovial membrane lining bed thickness of Normal group (A) is 1～2 layer, and cell arrangement is neat, and without inflammatory cell infiltration and blood vessel hyperplasia, articular cartilage is smooth without destroying.And the lymphocyte of adjurant arthritis rat hyperplasia of CIA model group (B) is obvious, arrangement disorder, synovial membrane backing layer cell is layer even more by 1～3 layer of original hypertrophy to 5～6, follow cellulose to ooze out, the collagen fiber calmness, visible a large amount of fusiformis fibroblast-like cells, synovial membrane thickens, synovial tissue is mushroom sample or nipple sample hypertrophy in various degree, the little blood vessel of synovial membrane lower floor increases, and pannus forms, and organizes lax, edema, have a large amount of inflammatory cells to ooze out, some section can see that synovial membrane grows to the cartilage surface aggressivity.Ethyl acetate low dose group (D) synovial tissue's moderate hypertrophy and inflammatory cell infiltration, have pannus to form and the minority cartilage destruction; The visible synovial tissue of dosage group (E) moderate hypertrophy in ethyl acetate, a small amount of inflammatory cell infiltration, without obvious cartilage and bone destruction; Ethyl acetate high dose group (F) synovial membrane is slight hypertrophy only, the cellular morphology rule, and a small amount of inflammatory cell infiltration, form without typical pannus, and cartilage surface is smooth, without cartilage destruction and bone erosion.The low dose group pathological changes changes and alleviates to some extent than model group, but pathological changes alleviates degree not as good as height, middle dosage group.Deformation failure, the paraplasm of synovial tissue, inflammatory cell infiltration that pharmaceutical composition of the present invention alleviates ankle joint are dose dependent reduction (seeing Fig. 6-14).
The swollen joint expansibility of 4 rats: the results are shown in Table 5.After treatment, each treatment group, tripterygium glycosides group are lower than model group (P<0.01 or P<0.05), and high dose group is lower than other each groups (P<0.O1 or P<0.05).
The impact of each group of table 5 on rat swollen joint expansibility
Each group on the CIA Rat Right after the impact of sole of the foot swelling
Annotate: with the blank group, compare,
#P ﹤ 0.05,
##P ﹤ 0.01; With model group, compare,
△P ﹤ 0.05,
△ △P ﹤ 0.01;
The content of 5 rat blood serum TNF-α, interleukin-6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10): the results are shown in Table 6.Model group rat blood serum IL-6, IL-8 and TNF-α and normal group is apparent in view increases (P<0.01), ethyl acetate and n-butyl alcohol hang down the content that group, middle group, high group, tripterygium glycosides group all can be lowered CIA rat blood serum IL-6, IL-8 and TNF-α, and each group of ethyl acetate is more variant significance (P<0.05).The high group of ethyl acetate is hanged down group aspect IL-6, IL-8 and TNF-α, more variant (P<0.05) with ethyl acetate.
Table 6 respectively organizes IL-6 in rat blood serum, IL-8 and the TNF-alpha content compares
Annotate: with normal group, compare,
*P ﹤ 0.05,
*P ﹤ 0.01; With model group, compare,
△P ﹤ 0.05,
△ △P ﹤ 0.01; With low dose group, compare,
#P<0.05.
3 discuss
The antiinflammatory experimentation shows, ethyl acetate and n-butyl alcohol pharmaceutical composition all can significantly alleviate the mice ear that dimethylbenzene causes, alleviates the rat toes swelling due to carrageenin, has antiinflammatory action preferably.
Analgesic experiment research shows, in hot plate method, before three dosage of pharmaceutical composition of the present invention and administration, obvious difference is arranged more all, can obviously extend the latency of pain response of mice, and along with the increase analgesic activity of dosage strengthens, effectively analgesic activity is lasting, more than maintaining 2h.
Simultaneously, pharmaceutical composition of the present invention can effectively be treated II Collagen Type VI arthritis, shows rheumatoid arthritis is had to good therapeutical effect.
Claims (9)
1. Caulis Bauhiniae Championii n-butanol extract, it is characterized in that: the weight percentage that it contains cupreol must not be less than 0.1%.
2. Caulis Bauhiniae Championii n-butanol extract according to claim 1, it is characterized in that: the weight percentage that it contains cupreol is 0.1-1.1%.
3. Caulis Bauhiniae Championii n-butanol extract according to claim 1 is characterized in that: it contain Quercetin,
4. according to the described Caulis Bauhiniae Championii n-butanol extract of claim 1-3 any one, it is characterized in that: it is to be prepared from by following method:
Get the Caulis Bauhiniae Championii medical material, add the 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol to nothing alcohol flavor, and the alcohol extract petroleum ether extraction, obtain petroleum ether layer and water layer, and water layer adds the chloroform extraction, obtains chloroform layer and water layer; The water layer ethyl acetate extraction, obtain ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, and obtains the Caulis Bauhiniae Championii n-butanol extract.
5. Caulis Bauhiniae Championii n-butanol extract according to claim 4, it is characterized in that: described ethanol is that concentration is 80% ethanol.
6. method for preparing the described Caulis Bauhiniae Championii n-butanol extract of claim 1-5 any one, it comprises the steps:
Get the Caulis Bauhiniae Championii medical material, add the 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol to nothing alcohol flavor, and the alcohol extract petroleum ether extraction, obtain petroleum ether layer and water layer, and water layer adds the chloroform extraction, obtains chloroform layer and water layer; The water layer ethyl acetate extraction, obtain ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, and obtains the Caulis Bauhiniae Championii n-butanol extract.
7. the purposes of the described Caulis Bauhiniae Championii n-butanol extract of claim 1-5 any one in the medicine for preparing antalgic and inflammation relieving, resisting rheumatoid arthritis.
8. pharmaceutical composition for the treatment of Rheumatoid arthritis, it is to be active component by the described Caulis Bauhiniae Championii n-butanol extract of claim 1-5 any one, adds pharmaceutically acceptable adjuvant or complementary composition to be prepared into preparation pharmaceutically commonly used.
9. the pharmaceutical composition for the treatment of rheumatoid arthritis according to claim 8, it is characterized in that: described preparation is oral formulations.
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| CN105726402B (en) * | 2016-03-22 | 2019-10-01 | 柳州两面针股份有限公司 | Dracaena boisiana extract is preparing the application in oral care product |
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-
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Non-Patent Citations (6)
| Title |
|---|
| 白海云等: "九龙藤化学成分研究(Ⅰ) ", 《中国中药杂志》 * |
| 白海云等: "九龙藤化学成分研究(Ⅰ)", 《中国中药杂志》, vol. 30, no. 01, 20 January 2005 (2005-01-20), pages 42 - 43 * |
| 白海云等: "九龙藤化学成分研究(Ⅱ) ", 《天然产物研究与开发》 * |
| 白海云等: "九龙藤化学成分研究(Ⅱ)", 《天然产物研究与开发》, vol. 16, no. 04, 30 August 2004 (2004-08-30), pages 312 - 313 * |
| 陆蕴如: "《中药化学》", 31 May 1995, 学苑出版社 * |
| 高杰等: "九龙藤乙酸乙酯提取物清除自由基、镇痛抗炎作用研究", 《安徽农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110787208A (en) * | 2018-11-20 | 2020-02-14 | 福建中医药大学 | Bauhinia championii total triterpene extract and preparation method and application thereof |
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