CN103405494B - 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., and also known as Caulis Bauhihiae Championii, Flos Mume nuphar japonicum, mainly distribute Fujian, Jiangxi, Guangxi, Zhejiang etc., has effect of wind-damp dispelling, promoting the circulation of blood gas.Modern pharmacology research proves, the effect that the effective ingredient of Caulis Bauhiniae Championii has expelling wind and removing dampness, promoting blood circulation and stopping pain, spleen invigorating are regulated the flow of vital energy, 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, be that the Cannizzaro method of main component has the effect of good treatment burn and scald clinically with Caulis Bauhiniae Championii.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 andanalgesic effect of Kangmeifu BurnsOintment (KBO) [J] .Chin J Tradit Med SciTechnol (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 BurnsOintment (KBO) [J] .Chin J Microecol (Chinese microecology magazine), 2007, 19 (2): 177-178.] report flavones ingredient in Caulis Bauhiniae Championii and there is 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 bibliographical information is had 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 volume the 1st phase, in January, 2005, to report from ethanol extraction isolation identification 5 compounds, its Extraction and separation method is: get Kowloon rattan 7Kg, with 95% industrial alcohol percolation 3 times after pulverizing.Extractum water suspendible, uses petroleum ether, ethyl acetate, n-butanol extraction successively.Ethyl acetate extract is separated with reversed-phase silica gel column chromatography through silica gel, macroporous resin, polydextran gel repeatedly, gets 5 compounds.
Bai Haiyun etc., Caulis Bauhihiae Championii chemical constitution study (II), Natural products research developed, 2004, Vol.16NO.4, be separated from ethyl acetate effective site and obtain 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 there is provided a kind of Caulis Bauhiniae Championii ethyl acetate extract, n-butanol extract.Another technical scheme of the present invention there is provided 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 prepared from by following steps:
Get Caulis Bauhiniae Championii medical material, add 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol extremely without alcohol taste, alcohol extract petroleum ether extraction, and obtain petroleum ether layer and water layer, water layer adds chloroform extraction, obtains chloroform layer and water layer; Aqueous layer with ethyl acetate extracts, and obtains acetic acid ethyl acetate extract, reclaims, and dries, obtains Caulis Bauhiniae Championii ethyl acetate extract.
Wherein, described ethanol to be concentration be 80% ethanol.
Present invention also offers a kind of method preparing described Caulis Bauhiniae Championii ethyl acetate extract, comprise the steps:
Get Caulis Bauhiniae Championii medical material, add 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol extremely without alcohol taste, alcohol extract petroleum ether extraction, and obtain petroleum ether layer and water layer, water layer adds chloroform extraction, obtains chloroform layer and water layer; Aqueous layer with ethyl acetate extracts, and obtains acetic acid ethyl acetate extract, reclaims, and dries, obtains Caulis Bauhiniae Championii ethyl acetate extract.
Present invention also offers the purposes of Caulis Bauhiniae Championii ethyl acetate extract in the medicine preparing antalgic and inflammation relieving, resisting rheumatoid arthritis.
The invention provides a kind of bauhinia championii n-butyl alcohol extract, the weight percentage that it contains cupreol must not be less than 0.1%.
Wherein, the weight percentage that it contains cupreol is 0.1-0.85%.
Further, it contains compound V, VI.
Bauhinia championii n-butyl alcohol extract of the present invention is prepared from by following method:
Get Caulis Bauhiniae Championii medical material, add 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol extremely without alcohol taste, alcohol extract petroleum ether extraction, and obtain petroleum ether layer and water layer, water layer adds chloroform extraction, obtains chloroform layer and water layer; Aqueous layer with ethyl acetate extracts, and obtains ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, obtains bauhinia championii n-butyl alcohol extract.Further preferably, described ethanol to be concentration be 80% ethanol.
Present invention also offers a kind of method preparing described bauhinia championii n-butyl alcohol extract, it comprises the steps:
Get Caulis Bauhiniae Championii medical material, add 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol extremely without alcohol taste, alcohol extract petroleum ether extraction, and obtain petroleum ether layer and water layer, water layer adds chloroform extraction, obtains chloroform layer and water layer; Aqueous layer with ethyl acetate extracts, and obtains ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, obtains bauhinia championii n-butyl alcohol extract.
Present invention also offers the purposes of this bauhinia championii n-butyl alcohol extract in the medicine 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 stablized, and are applicable to large production, provide a kind of selection newly for clinical.
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
Detailed description of the invention
The component separating of embodiment 1 Caulis Bauhiniae Championii ethyl acetate extract and n-butanol portion and purification
1. Extraction and separation
Get 1000g Caulis Bauhiniae Championii medical material, add 80% alcohol heating reflux and extract 3 times, each 1.5h, filter, merge ethanol, reclaim ethanol to without alcohol taste, obtain alcohol-extracted extract.Add 2000ml water, make extractum suspendible, add petroleum ether extraction 5 times, 1000ml/ time, merge petroleum ether extract, reclaim, obtain grease; Water layer adds chloroform extraction 5 times, 1000ml/ time, combined chloroform extracting solution, reclaims, and dries, obtains extractum; Water layer adds extraction into ethyl acetate 5 times, 1000ml/ time, combined ethyl acetate extracting solution, reclaims, and dries, obtains extractum; Water layer adds n-butanol extraction 5 times, 1000ml/ time, merges n-butanol extracting liquid, reclaims, and dries, obtains extractum.
The Caulis Bauhiniae Championii ethyl acetate extract that this method obtains calculates by extractum, and yield must not be less than 2%, must not be less than 0.05% and 0.04% respectively containing Quercetin and ampelopsin.
The Caulis Bauhiniae Championii n-butanol portion that this method obtains calculates by extractum, and yield must not be less than 6%, must not be less than 0.1% respectively containing cupreol.
Concrete assay method is:
HPLC method is adopted to measure the content of Quercetin, ampelopsin in Caulis Bauhiniae Championii ethyl acetate extract.Chromatographic condition is acetonitrile (A)-0.05% phosphoric acid solution (B), gradient elution program be 0-5min acetonitrile from 20% to 30%, 5-10min from 30% to 50%, then keep 20min; Flow velocity is 1.0ml/min; Mensuration wavelength is 370nm; Column temperature is 30 DEG C.
Test in triplicate, method measures 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%.
HPLC method is adopted to measure the content of cupreol in bauhinia championii n-butyl alcohol extract.Chromatographic condition is acetonitrile (A)-water (B) (96:4); Flow velocity is 1.0ml/min; Mensuration wavelength is 210nm; Column temperature is 30 DEG C.
Test in triplicate, method measures the content that extract obtains cupreol and is respectively 0.364%, 0.363%, 0.363% thus.Average content is 0.363%.
Ethyl acetate portion thick silica gel (160 ~ 200 order) 1:1 mixes sample, add with in dichloromethane wet method dress silicagel column, use methylene chloride-methanol gradient elution, equal-volume collects eluent, flow point is examined knowledge through TLC and is remerged, through silica gel repeatedly, polyamide, SephadexLH-20, ODS abstraction and purification, obtain compounds I, II, III, IV, V.
N-butanol fraction is with hot water dissolving, the insoluble position of water thick silica gel (160 ~ 200 order) 1:1 mixes sample, add with in dichloromethane wet method dress silicagel column, use methylene chloride-methanol gradient elution, equal-volume collects eluent, flow point is examined knowledge through TLC and is remerged, and through silica gel repeatedly, polyamide, SephadexLH-20, ODS abstraction and purification, obtains compound V, VI.
2. Structural Identification
Compounds I: white, needle-shaped crystals (acetone), mp141-142 DEG C.5% sulphuric acid-alcoholic solution heating displaing amaranth, vanillin-concentrated sulphuric acid displaing amaranth.Be presented at 3432 in IR, 2936,2860,1461,1381,1054,959,800cm
-1there is absorption band at place, and the matching degree of composing cupreol in storehouse with Sigma Biological Sample Library reaches more than 90%.Under petroleum ether-ethyl acetate, petroleum ether-acetone, dichloromethane-acetone three kinds of different unfolding conditions, its Rf value is consistent with sitosterol, and mixed melting point does not decline.Consistent with document, therefore be accredited as cupreol, structural formula is shown in Fig. 1.
Compound ii: white powder, mp288 ~ 289 DEG C.Be presented at 3397 in IR, 2959,2933,2869,1463,1379,1073,1024cm
-1there is absorption band at place, and the matching degree of composing daucosterol in storehouse with Aldrich Condensed PhaseSample Library reaches more than 90%.Silica gel thin-layer plate launches by systems such as dichloromethane-acetone, methylene chloride-methanol, methylene chloride-methanol-water, and be single-point, its Rf value is consistent with daucosterol standard substance, and mixed melting point does not decline, therefore is accredited as daucosterol, and structural formula is shown in Fig. 2.
Compound III: white flaky crystals (petroleum ether-ethyl acetate), mp65 ~ 66 DEG C.Be presented at 2917 in IR, 2848,1472,1061,719cm
-1there is absorption band, the matching degree of composing melissane in storehouse with Aldrich CondensedPhase Sample Library reaches more than 90%, and launches with different dicyandiamide solutions on silica gel thin-layer plate, is single-point, its Rf value is consistent with melissane standard substance, therefore is accredited as positive melissane.
Compounds Ⅳ: white flaky solid (petroleum ether-ethyl acetate), mp97 ~ 98 DEG C.Be presented at 2917 in IR, 2849,1742,1472,1363,1239,1179cm
-1there is absorption band at place, and the matching degree of composing hexacontane in storehouse with Aldrich Condensed Phase Sample Library reaches more than 90%, and launches with different dicyandiamide solutions on silica gel thin-layer plate, is single-point, therefore is accredited as positive hexacontane.
Compound V: yellow needles (chloroform-methanol), mp > 300 DEG C.Hydrochloric acid-magnesium powder reacting positive.Be presented at 2918 in IR, 2849,1664,1610,1521,1450,1381,1318,1261,1198,1167,1013cm
-1there is absorption band at place, and the matching degree of composing Quercetin in storehouse with HR Aldrich Aldehydes and Ketones reaches more than 90%, 1H-NMR data and bibliographical information
[7]quercetin data consistent.And launch with different dicyandiamide solutions on silica gel thin-layer plate, be single-point, and Rf value is consistent with Quercetin reference substance, mixed melting point does not decline.Therefore being accredited as Quercetin, structural formula is shown in Fig. 3.
Compound VI: yellow particle shape crystal (chloroform-methanol), mp > 280 DEG C.Be presented at 3409,1647,1611,1576,1458,1293,1257,1168,1119cm-1 in IR and have absorption band, through hydrogen spectrum, Mass Spectrometric Identification, infer that the molecular weight of compound is: 326, preliminary putative structure is as Fig. 4.
Beneficial effect of the present invention is proved below by way of concrete pharmacodynamics test.
Medicine and reagent:
Tripterygium wilfordii Polyglycosidium Tablets, Huangshi Feiyun Pharmaceutical Co., Ltd. produces (authentication code:, the accurate word Z42021212 of traditional Chinese medicines);
Medicine of the present invention (Caulis Bauhiniae Championii ethyl acetate extract, n-butanol extract prepared by embodiment 1); The low dosage of medicine of the present invention 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 medicine inspecting institute of Fujian Province, is made into 1% with normal saline, uses after placing 2d.Sodium sulfide: Shanghai Tong Ya chemical science and technology 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, is produced, purchased from Bo Leide bio tech ltd, Beijing by SIGMA company of the U.S..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, 8 groups are divided at random: low dosage medicine group of the present invention by body weight, 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 mices gavage respectively, every day 1 time, continuous 3d, after last coating 30min, smear at the allow uniform that auris dextra both sides are identical with 0.1ml/ amount only with dimethylbenzene, left ear compares, put to death after 4h, cut ears immediately, same area auricle is got with diameter 8mm card punch, ten thousand/semi-automated analysis balance is weighed, presses with (auris dextra weight-left ear weight)/left ear heavily for swelling rate, with (the average swelling of model group-average swelling of administration group) average swelling of/model group for inhibitory rate of intumesce.The swelling degree of matched group and administration group is used SPSS statistical software Treatment Analysis, and between its average, difference adopts one factor analysis of variance, in table 1.
Table 1 medicine group of the present invention is on the impact (n=10) of mice caused by dimethylbenzene xylene inflammation
Compare with model control group, 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 is all greater than 20%, points out medicine group xylol induced mice auricle edema of the present invention inhibited.And in dose-effect relationship.
2 on Carrageenan cause the impact of rat paw edema: by 80 body weight 150 ± 10g gram SD rat, be divided into 8 groups at random: model group by body weight, the basic, normal, high dosage group of medicine of the present invention, Tripterygium wilfordii Polyglycosidium Tablets group.Rat right hind leg is stretching, subcutaneous injection 1% carrageenin 0.1mL in the middle part of the self-sustaining sole of the foot.Respectively at the thickness injecting rear 0.5h, lh, 1.5h, 2h, 4h, 6h vernier caliper measurement swollen limb, with (after causing inflammation-cause scorching front foot sole of the foot thickness)/cause scorching front foot sole of the foot thickness × 100% to calculate swelling rate, suppression ratio is calculated with average swelling rate × 100% of (the average swelling rate of the average swelling rate-experimental group of model control group)/model control group, each group of date processing 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 to rat carrageenan
Result shows, each medicine group more all has significant difference (P<0.05) in causing scorching rear 1h to 6h with model group, points out rat paw edema caused by each medicine group on Carrageenan all to have inhibitory action.Three dosage of medicine group of the present invention, after causing inflammation, 1h suppression ratio just reaches more than 30%; Peak during 1h, 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 middle high dose reaches more than 6h, carries out one factor analysis of variance show when 4h and 6h, basic, normal, high dosage and Tripterygium wilfordii Polyglycosidium Tablets no significant difference (P>0.05).
The analgesic experiment (impact on female mice hot plate induced pain) of test example 2 medicine of the present invention
Get the female mice of body weight 20 ± 2g, before test, on hot plate dolorimeter, measure the mice of pain threshold in 5s-30s 80 by hot plate method, be divided into 8 groups at random by body weight: blank group, the basic, normal, high dosage group of medicine group of the present invention, Tripterygium wilfordii Polyglycosidium Tablets group.To occur licking metapedes reaction for observation index.Respectively at the above medicine of every Mus gavage, 0.2mL/ only.Measure pain threshold by upper method after 0.5h, then by front method administration, measure the pain threshold of 1h, 1.5h, 2h.Compare the change of the threshold of pain before and after administration, and press (after medication-and the front average pain threshold of medication)/medication front on average pain threshold × 100% calculating threshold of pain raising percentage rate.Each group of date processing uses SPSS statistical software Treatment Analysis, and between its average, difference adopts one factor analysis of variance, the results are shown in Table 3.
Table 3 medicine is on the impact (n=10) of mouse hot-plate induced pain threshold and threshold of pain increase rate
Result shows, and pharmaceutical composition of the present invention compares with model control group significant difference (P<0.O5), and still has analgesic effect during 2h.Pharmaceutical composition of the present invention three dosage all can significantly improve mice pain threshold.Prompting: pharmaceutical composition of the present invention has remarkable analgesic activity to thermic pain, and analgesia reaches more than 2h lastingly.
Test example 3 medicine of the present invention is on arthritic impact
Grouping and modeling: Wistar rat (male), average weight (200 ± 20) g, adaptability is raised 1 week, and unified illumination, low temperature environment, freely drinks water, and solid feed is freely ingested.Be divided into 9 groups at random, i.e. 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, often group is 10.Mixed with the cattle II Collagen Type VI emulsion equal-volume of 2mg/ml by complete Freund's adjuvant (CFA), make collagen emulsion, except normal group, every rat root of the tail portion intradermal injection collagen protein 0.2mg, namely after initial immunity the 1st day start the gastric infusion that divides into groups, every day 1 time.Carried out secondary immunity in the 7th day, except normal group, every rat root of the tail portion 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; 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 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 and measure rats with bilateral foot pawl volume, record volumetric values, in this, as the radix evaluating antiinflammatory action intensity.
Sample collection and Indexs measure: respectively before molding, survey the change of pedal swelling thickness and body weight after modeling; Before treatment, after treatment, calculate arthritis swelling; After treating administration in the 28th day, all animals abdominal aortic blood, dividing is put in drying tube, drying tube is placed in 3000 revs/min of centrifugal 8min, get supernatant separation of serum, press the content that test kit description method measures TNF-α, interleukin-6 (IL-6), interleukin 8 (IL-8), interleukin 10 (IL-10) respectively.
Statistical method: adopt SPSS15.0 data statistics bag to carry out date processing, experimental data is used
represent.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 model group rats is after collagen emulsion sensitization, booster immunization, and the mental status, diet situation, mobility decrease, and after initial immunity, festering appears in afterbody, forms a scab gradually after 7 days.After secondary immunity about the 3rd day, rat started morbidity, and rat model toes and ankle joint start to occur swelling, and day by day increase the weight of, weight loss, and the right back foot of partial rat occurs that toes be out of shapes, without death; Positive controls and the rear above-mentioned symptom of Caulis Bauhiniae Championii group rat treatment are obviously alleviated.
The body weight of 2 rats: the results are shown in Table 4.Except medicine group, other are respectively organized rat body weight and slowly increase, and during 14 days all 21 days, body weight starts to decline, and then slowly increases.
Table 4 each group of impact on rat body weight
Note: compare with normal group,
#p ﹤ 0.05,
##p ﹤ 0.01; Compare with model group,
*p ﹤ 0.05,
*p ﹤ 0.01;
3 pathological study: each treated animal ankle joint pathology display, the synovial cells in rats synovial membrane lining 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 even more to 5 ~ 6 layers by original 1 ~ 3 layer of hypertrophy, ooze out with cellulose, collagen fiber are calm, visible a large amount of fusiformis fibroblast-like cells, synovial membrane thickens, synovial tissue is in mushroom sample in various degree or nipple sample hypertrophy, the thin vessels of synovial membrane lower floor increases, and pannus is formed, scrim, edema, have a large amount of inflammatory cell to ooze out, some section can see that synovial membrane grows to cartilage surface aggressivity.Ethyl acetate low dose group (D) synovial tissue moderate hypertrophy and inflammatory cell infiltration, have pannus to be formed and minority cartilage destruction; Dosage group (E) visible synovial tissue 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, cellular morphology rule, and a small amount of inflammatory cell infiltration, formed without typical blood vessel nebula, cartilage surface is smooth, without cartilage destruction and bone erosion.Low dose group pathological changes changes comparatively model group and alleviates to some extent, but pathological changes alleviates degree not as good as height, middle dosage group.Pharmaceutical composition of the present invention alleviates the deformation failure of ankle joint, the paraplasm of synovial tissue, inflammatory cell infiltration are that dose dependent reduces (see Fig. 6-14).
The Articular swelling 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 group (P<0.O1 or P<0.05).
Table 5 each group of impact on rat articular swelling
The each group of impact on sole of the foot swelling after CIA Rat Right
Note: compare with blank group,
#p ﹤ 0.05,
##p ﹤ 0.01; Compare with model group,
△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 rats blood serum IL-6, IL-8 and TNF-α and normal group are obvious to be increased (P<0.01), ethyl acetate and low group of n-butyl alcohol, middle group, high group, tripterygium glycosides group all can lower the content of CIA rat blood serum IL-6, IL-8 and TNF-α, ethyl acetate respectively organizes more variant significance (P<0.05).Ethyl acetate height is organized with low group of ethyl acetate in IL-6, IL-8 and TNF-α, more variant (P<0.05).
Table 6 is respectively organized IL-6, IL-8 and TNF-alpha content in rat blood serum and is compared
Note: compare with normal group,
*p ﹤ 0.05,
*p ﹤ 0.01; Compare with model group,
△p ﹤ 0.05,
△ △p ﹤ 0.01; Compare with low dose group,
#p<0.05.
3 discuss
Antiinflammatory experimentation shows, ethyl acetate and n-butyl alcohol pharmaceutical composition all significantly can alleviate the mice ear that dimethylbenzene causes, and alleviates the rat toes swelling caused by carrageenin, has good antiinflammatory action.
Analgesic experiment research shows, in hot plate method, pharmaceutical composition of the present invention three dosage more all have obvious difference with before administration, obviously can extend the latency of pain response of mice, and strengthen along with the increase analgesic activity of dosage, effective analgesic activity is lasting, can maintain more than 2h.
Meanwhile, pharmaceutical composition of the present invention effectively can treat II Collagen Type VI arthritis, shows to have good therapeutical effect to rheumatoid arthritis.
Claims (8)
1. a bauhinia championii n-butyl alcohol extract, is characterized in that: the weight percentage that it contains cupreol must not be less than 0.1%, and described bauhinia championii n-butyl alcohol extract is prepared from by following steps:
Get Caulis Bauhiniae Championii medical material, add 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol extremely without alcohol taste, alcohol extract petroleum ether extraction, and obtain petroleum ether layer and water layer, water layer adds chloroform extraction, obtains chloroform layer and water layer; Aqueous layer with ethyl acetate extracts, and obtains ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, obtains bauhinia championii n-butyl alcohol extract.
2. bauhinia championii n-butyl alcohol extract according to claim 1, is characterized in that: the weight percentage that it contains cupreol is 0.1-1.1%.
3. bauhinia championii n-butyl alcohol extract according to claim 1 and 2, is characterized in that: it contain Quercetin,
4. bauhinia championii n-butyl alcohol extract according to claim 1, is characterized in that: described ethanol to be concentration be 80% ethanol.
5. prepare a method for bauhinia championii n-butyl alcohol extract according to claim 1, it comprises the steps:
Get Caulis Bauhiniae Championii medical material, add 40-95% alcohol heating reflux and extract, extracting solution reclaims ethanol extremely without alcohol taste, alcohol extract petroleum ether extraction, and obtain petroleum ether layer and water layer, water layer adds chloroform extraction, obtains chloroform layer and water layer; Aqueous layer with ethyl acetate extracts, and obtains ethyl acetate layer and water layer; Water layer adds n-butanol extraction, obtains n-butanol extracting liquid, reclaims, and dries, obtains bauhinia championii n-butyl alcohol extract.
6. the purposes of bauhinia championii n-butyl alcohol extract according to claim 1 in the medicine preparing antalgic and inflammation relieving, resisting rheumatoid arthritis.
7. treat a pharmaceutical composition for Rheumatoid arthritis, it is active component by bauhinia championii n-butyl alcohol extract according to claim 1, adds pharmaceutically acceptable adjuvant or complementary composition is prepared into pharmaceutically conventional preparation.
8. the pharmaceutical composition for the treatment of rheumatoid arthritis according to claim 7, is characterized in that: described preparation is oral formulations.
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| CN201310289110.XA CN103405494B (en) | 2012-03-31 | 2012-03-31 | Bauhinia championii n-butyl alcohol extract and preparation method and application thereof |
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| 九龙藤乙酸乙酯提取物清除自由基、镇痛抗炎作用研究;高杰等;《安徽农业科学》;20111231;第39卷(第36期);22305-22306 * |
| 九龙藤化学成分研究(Ⅰ);白海云等;《中国中药杂志》;20050120;第30卷(第01期);42-43 * |
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