CN105326872A - Preparation method for herba patriniae extract with therapeutical effect on ulcerative colitis - Google Patents
Preparation method for herba patriniae extract with therapeutical effect on ulcerative colitis Download PDFInfo
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Abstract
The invention discloses a preparation method for a herba patriniae extract with therapeutical effect on ulcerative colitis. The preparation method comprises the following steps: crushing herba patriniae, adding water to leach the herba patriniae, and concentrating the herba patriniae; extracting concentrated solution with water saturated n-butyl alcohol, removing a solvent to obtain residues, dissolving the residues with water, absorbing by use of polyamide resin, eluting by water, collecting hydrolysate, and drying the hydrolysate to obtain dry powder; dissolving the dry powder, adsorbing by use of macroporous adsorption resin, eluting by water, discarding the hydrolysate, eluting by alcohol, collecting eluant, concentrating and drying the eluant to obtain the herba patriniae extract. The herba patriniae extract extract obtained by the method is used as the only active ingredient, the SOD value of a rat with ulcerative colitis can be increased, an MDA value is reduced, and MPO activity is reduced, so that oxygen radicals are removed; lipid peroxidase is reduced, neutrophil infiltration is reduced and colitis symptoms are improved or relieved. Meanwhile, the herba patriniae extract can reduce TNF-alpha and IL-1beta level, is obvious in improvement on rat ulcerative colitis and is expected to be developed into a novel efficient ulcerative colitis treatment drug.
Description
Technical field
The application of Herba Patriniae extract that the present invention relates to a kind of preparation method to the medicative Herba Patriniae extract of ulcerative colitis tool and prepared by the method.
Background technology
Herba Patriniae is Valerianaceae herbaceos perennial Patrinia scabiosaefolia Fisch, the rhizome of patrima villosa or whole herb with root, is Chinese medicine ancient and conventional simply.Herba Patriniae has effect of heat-clearing and toxic substances removing, evacuation of pus removing blood stasis with potent drugs, blood circulation promoting and blood stasis dispelling.Clinically be usually used in chronic gastritis, gastroesophageal reflux, radiation rectitis etc.The existing modern pharmacological research reports of effect such as antibacterial, the antiviral of Herba Patriniae extract, blood fat reducing and CNS inhibition, but have no and be used alone the report of Herba Patriniae to treatment of ulcerative colitis aspect.
Existing Chinese medicine extract, generally lacks the extracting method of science, and still extract with conventional methods such as traditional decocting, ethanol extractions, the impurity content in medicine is many, and even containing harmful composition, this has had a strong impact on the curative effect of Chinese medicine extract.
Summary of the invention
The object of the present invention is to provide a kind of preparation method to the medicative Herba Patriniae extract of ulcerative colitis tool.
The technical solution used in the present invention is:
A preparation method for Herba Patriniae extract, comprises the steps:
1) Herba Patriniae is pulverized, add flooding, lixiviating solution is concentrated, obtain concentrated solution;
2) concentrated solution water-saturated n-butanol is extracted, extract is merged, remove solvent, obtain residue;
3) by residue water dissolution, solution polyamide is adsorbed, washes with water, collect water lotion, dry, obtain dry powder;
4) dry powder is dissolved, by solution absorption with macroporous adsorbent resin, wash with water, abandon water lotion, then with 30 ~ 95% ethanol elutions, collect ethanol elution, namely concentrate drying obtains Herba Patriniae extract.
As the further improvement of above-mentioned preparation method, the preparation manipulation of concentrated solution is: pulverized by Herba Patriniae, add the water soaking of medical material volume 2 ~ 5 times amount, soak time 0.5 ~ 2h, use the water extraction 1 ~ 3 time of quality of medicinal material 6 ~ 14 times amount again, extract 0.5 ~ 2 hour, merge extractive liquid, at every turn, filter, filtrate is condensed into the concentrated solution that concentration is the former medicine/mL of 0.5 ~ 1g.
As the further improvement of above-mentioned preparation method, the preparation manipulation of residue is: the water-saturated n-butanol getting concentrated solution 1 ~ 3 times of volume extracts 1 ~ 3 time repeatedly, merges butanol extraction liquid, and recycling design is to dry.
As the further improvement of above-mentioned preparation method, the amount ratio of residue and polyamide is: the former medicine of 1g/(1 ~ 3) g polyamide.
As the further improvement of above-mentioned preparation method, the amount ratio of dry powder and macroporous adsorbent resin is: the former medicine of 1g/(1 ~ 3) g macroporous adsorbent resin.
As the further improvement of above-mentioned preparation method, the model of macroporous adsorbent resin is the one in D101, D140, D141, AB-8.
As the further improvement of above-mentioned preparation method, preparation method comprises the steps:
1) Herba Patriniae is pulverized, add the water soaking of medical material volume 2 ~ 3 times amount, soak time 0.5 ~ 2h, use the water extraction 1 ~ 2 time of quality of medicinal material 8 ~ 12 times amount again, extract 1 ~ 1.5 hour, merge extractive liquid, at every turn, filter, filtrate is condensed into the concentrated solution that concentration is 0.6 ~ 0.9 former medicine g/mL;
2) water-saturated n-butanol getting concentrated solution 1 ~ 2 times of volume extracts 1 ~ 3 time repeatedly, merges butanol extraction liquid, and recycling design, to dry, obtains residue;
3) get the solution that residue water dissolution becomes the former medicine/mL of 0.6 ~ 0.9g, solution polyamide is adsorbed, washes with water, collect water lotion, dry, obtain dry powder;
4) dry powder is dissolved into the solution of the former medicine/mL of 0.6 ~ 0.9g, by solution absorption with macroporous adsorbent resin, washes with water, abandon water lotion, then with 55 ~ 75v/v% ethanol elution, collect ethanol elution, namely concentrate drying obtains Herba Patriniae extract.
Treat a compositions for ulcerative colitis, its active component is Herba Patriniae extract, and the preparation method of Herba Patriniae extract is described above.
Preferably, the dosage form of compositions is oral formulations.
Preferably, oral formulations is selected from tablet, capsule, granule, drop pill.
The invention has the beneficial effects as follows:
The inventive method extracts the Herba Patriniae extract obtained, main containing saponin components such as scabioside A, scabioside B, scabioside C, oleanolic acid, α-Hederagenin, β-Hederagenins, and the total content of these effective ingredient is between 50% to 100%.This Herba Patriniae extract, as unique active component, can raise ulcerative colitis in rats SOD value, reduces MDA value, reduce MPO active, thus scavenging activated oxygen, reduce over lipid oxide and generate, reduce neutrophil infiltration, improve or alleviate colitis symptoms.TNF-α, IL-1 β level can be reduced simultaneously, thus the rat colonitis of TNBS or Ethanol Method induction is improved significantly, be that prior art cannot realize, expand the Herba Patriniae scope of application.Be expected to be developed as novel efficient treatment of ulcerative colitis medicine.
Detailed description of the invention
Below in conjunction with embodiment, further illustrate technical scheme of the present invention.
The Herba Patriniae medical material used in following examples is purchased from peaceful medical material market, Guangzhou.
Embodiment 1
1) pulverized by Herba Patriniae, add the water soaking of medical material volume 2 times amount, soak time 0.5h, then use the water extraction 1 time of quality of medicinal material 12 times amount, extracts 1 hour, filtration, filtrate is condensed into the solution that concentration is 0.6g/mL (being equivalent to crude drug) concentration;
2) get the above-mentioned solution water-saturated n-butanol of 1 times of volume and extract 1 time, collect butanol extraction liquid, recycling design, to dry, obtains residue;
3) get the solution that residue water dissolution becomes 0.6g/mL (being equivalent to crude drug) concentration, by this solution polyamide with water elution, collect water lotion, be evaporated to dry, obtain dry powder;
4) get the solution that dry powder water dissolution becomes 0.6g/mL (being equivalent to crude drug) concentration, with D101 macroporous adsorbent resin with water elution, discard water lotion, again with 55% ethanol elution, collect ethanol elution, reclaim ethanol, thick paste is drying to obtain Herba Patriniae extract.
Total content that the present embodiment obtains saponin component in Herba Patriniae extract is 67% to utilize ultraviolet visible spectrophotometry (with scabioside C for reference substance) to record.
Embodiment 2
1) Herba Patriniae is pulverized, add the water soaking of medical material volume 3 times amount, soak time 1h, use the water extraction 2 times of quality of medicinal material 8 times amount again, to extract 1.5 hours at every turn, merge extractive liquid, filter, filtrate is condensed into the solution that concentration is 0.8g/mL (being equivalent to crude drug) concentration;
2) water-saturated n-butanol getting above-mentioned solution 2 times of volumes extracts 3 times repeatedly, merges butanol extraction liquid, and recycling design, to dry, obtains residue;
3) get the solution that residue water dissolution becomes 0.8g/mL (being equivalent to crude drug) concentration, by this solution polyamide with water elution, collect water lotion, be evaporated to dry, obtain dry powder;
4) get the solution that dry powder water dissolution becomes 0.8g/mL (being equivalent to crude drug) concentration, with AB-8 macroporous adsorbent resin with water elution, discard water lotion, again with 75% ethanol elution, collect ethanol elution, reclaim ethanol, thick paste is drying to obtain Herba Patriniae extract.
Total content that the present embodiment obtains saponin component in Herba Patriniae extract is 54% to utilize ultraviolet visible spectrophotometry (with scabioside C for reference substance) to record.
Embodiment 3
1) Herba Patriniae is pulverized, add the water soaking of medical material volume 2 times amount, soak time 2h, use the water extraction 2 times of quality of medicinal material 9 times amount again, to extract 1 hour at every turn, merge extractive liquid, filter, filtrate is condensed into the solution that concentration is 0.9g/mL (being equivalent to crude drug) concentration;
2) water-saturated n-butanol getting above-mentioned solution 1 times of volume extracts 3 times repeatedly, merges butanol extraction liquid, and recycling design, to dry, obtains residue;
3) solution that residue water dissolution becomes 0.9g/mL (being equivalent to crude drug) concentration is got.By this solution polyamide with water elution, collect water lotion, be evaporated to dry, obtain dry powder;
4) get the solution that dry powder water dissolution becomes 0.9g/mL (being equivalent to crude drug) concentration, with D140 macroporous adsorbent resin with water elution, discard water lotion, again with 65% ethanol elution, collect ethanol elution, reclaim ethanol, thick paste is drying to obtain Herba Patriniae extract.
Total content that the present embodiment obtains saponin component in Herba Patriniae extract is 72% to utilize ultraviolet visible spectrophotometry (with scabioside C for reference substance) to record.
Comparative example 1
1) pulverized by Herba Patriniae, add the water soaking of medical material volume 2 times amount, soak time 0.5h, then use the water extraction 1 time of quality of medicinal material 12 times amount, extracts 1 hour, filtration, filtrate is condensed into the solution that concentration is 0.6g/mL (being equivalent to crude drug) concentration;
2) get the above-mentioned solution water-saturated n-butanol of 1 times of volume and extract 1 time, collect butanol extraction liquid, recycling design, thick paste is drying to obtain Herba Patriniae extract.
Total content that the present embodiment obtains saponin component in Herba Patriniae extract is 18% (content is less than 50%) to utilize ultraviolet visible spectrophotometry (with scabioside C for reference substance) to record.
Comparative example 2
Pulverized by Herba Patriniae, add the water soaking of medical material volume 3 times amount, soak time 1h, then use the water extraction 2 times of quality of medicinal material 8 times amount, to extract 1.5 hours at every turn, merge extractive liquid, filter, filtrate is condensed into thick paste, dry, pulverize to obtain Herba Patriniae extract.
Total content that the present embodiment obtains saponin component in Herba Patriniae extract is 12% (content is less than 50%) to utilize ultraviolet visible spectrophotometry (with scabioside C for reference substance) to record.
Comparative example 3
Herba Patriniae is pulverized, adds 95% soak with ethanol of medical material volume 2 times amount, soak time 2h, then use 95% alcohol reflux 2 times of quality of medicinal material 9 times amount, each extraction 1 hour, merge extractive liquid, filters, filtrate recycling design, dry, pulverize to obtain Herba Patriniae extract.
Total content that the present embodiment obtains saponin component in Herba Patriniae extract is 8% (content is less than 50%) to utilize ultraviolet visible spectrophotometry (with scabioside C for reference substance) to record.
The preparation of Herba Patriniae Changning tablet
The Herba Patriniae extract prepared with embodiment 1 is to prepare Herba Patriniae Changning tablet, and preparation method is: take 250g Herba Patriniae extract, adds 250g calcium sulfate, the mixing of 10g polyvinylpolypyrrolidone.Make soft material with appropriate 80% ethanol as wetting agent, granulate with 14 mesh sieves, after dry at 60 DEG C of temperature, cross 12 mesh sieve granulate, after adding 10g polyvinylpolypyrrolidone and the mixing of 3g magnesium stearate, make 1000.
The Herba Patriniae extract prepared with embodiment 2 is to prepare Herba Patriniae Changning tablet, and preparation method is the same.
The Herba Patriniae extract prepared with embodiment 3 is to prepare Herba Patriniae intestinal Yiganning capsule, and preparation method is: take 250g Herba Patriniae extract, adds 250g calcium sulfate, the mixing of 10g polyvinylpolypyrrolidone.Make soft material with appropriate 80% ethanol as wetting agent, granulate with 14 mesh sieves, cross 12 mesh sieve granulate after dry at 60 DEG C of temperature, insert in capsulae vacuus, to obtain final product.
Herba Patriniae Changning tablet is to the protective effect of the rat colonitis tissue that trinitro-benzene-sulfonic acid is induced
Concrete steps are as follows:
Material and instrument: Herba Patriniae Changning tablet (BJC); Sulfasalazine (SASP, Hainan Hong Hui pharmaceutical Co. Ltd produces, and is made into the suspension of 0.3mg/mL before use with distilled water); 2,4,6-trinitro-benzene-sulfonic acid (TNBS, Sigma company); Pentobarbital sodium (Beijing chemical reagents corporation); MPO, SOD, MDA detection kit (Bioengineering Research Institute is built up in Nanjing).SPF level SD rat 60, weight 180 ~ 220g, provides by Guangdong Province's (medical science) Experimental Animal Center, quality certification SCXK (Guangdong) 2008-0002.U-2900 type is visible-ultraviolet spectrophotometer (HIT); KDC-220HR High speed refrigerated centrifuge (Zhong Jia branch company of Keda Innovation Co., Ltd); Star sea Rotary Evaporators (Wuxi City Xing Haiwang biochemical equipment company limited); 420 thermostat water baths (positive group Instrument Ltd. of Jintan City); XH-B vortex mixer (healthy Medical Devices Co., Ltd. of Jiangyan City); Glass homogenizer (Shanghai glass apparatus factory); AEL-160 electronic analytical balance (Japanese Shimadzu Corporation).
Test method:
Modeling with draw materials: 60 SD rats adapted to raising after 1 week, at random rat were divided into normal group, model group, SASP group (0.3g/kg), the high, medium and low dosage group of BJC (16,8,4g crude drug/kg), often group 10, water is can't help in fasting.After 24h 10% pentobarbital sodium 3.5mLkg
-1intraperitoneal anesthesia, the latex tubing per anum of diameter 2mm is inserted gently about 8cm place in rat body, with 50% alcoholic solution of the disposable injection TNBS of syringe to (TNBS125mg/kg) every 0.5mL in the enteric cavity of rat, the afterbody of rat is mentioned, be inverted 30s, make modeling agent fully infiltrate through the enteric cavity of rat, normal group injects equivalent 50% alcoholic solution.Start gastric infusion after modeling 24h, normal group and model group give normal saline, continue medication 4 weeks.After last administration 2h, rat cervical dislocation is put to death, open abdomen and be separated colon, cut off enteric cavity along mesentery edge, 4 DEG C of normal saline flushings, are laid on plastic plate, perusal Traumatic Colon degree, carry out colon general form Injury score, index comprises adhesion contrafluxion, ulcer and inflammation.Leave and take the colonic mucosal tissue apart from anus 8 ~ 10cm, paraffin embedding, section, dewaxing, hematoxylin-eosin (HE) dyes, dehydration of alcohol, the transparent mounting of dimethylbenzene, light Microscopic observation Injured colonic mucosa, carries out colon histology Injury score, and index comprises ulcer inflammatory granuloma, fibrosis and injured depth.It is to be measured that remaining rat colon organizes liquid nitrogen-70 DEG C to preserve.
Biochemical Indexes: get rat colon tissue, illustrates according to test kit and measures MPO, SOD, MDA content.
Data statistics processing method: adopt statistic software SPSS 13.0 to carry out one factor analysis of variance process, experimental result represents with mean ± standard deviation (), adopt rank test to general form and Histological injury's ranked data, P<0.05 is for having statistical significance.
Result of the test:
The impact of the ulcerative colitis in rats colon general form that BJC induces TNBS and Histological injury: the results are shown in Table 1.Normal rats is without suffering from diarrhoea, having blood in stool, and colonic mucosa is smooth without ulcer, body of gland marshalling.Model group (TNBS group) rat shows as dispirited, perpendicular hair, hogback, lazy dynamic, anorexia, all occurs diarrhoea in various degree, loose stool, with hemafecia, and sustainable more than 3 weeks.Animal is put to death after 4 weeks, visible colon and tissue adhesion, mucosa congestion and edema, ulcer, intestinal tube diseased region occurs narrow, visible ulcer and inflammatory exudate under light microscopic, and main manifestations is neutrophil infiltration, there is mucosa body of gland arrangement distortion, compare general form with normal group and to mark and histological score obviously increases (P<0.05).The high, medium and low dosage group of BJC and SASP group general form are marked and histological score obviously reduces, compare with model group (P<0.05), all can improve the inflammatory symptoms such as colon congestion and edema, reduce ulcer area, promote ulcer healing.
The impact of the ulcerative colitis in rats colon general form that table 1BJC induces TNBS and Histological injury (
)
| Group | Dosage C/gKg -1 | General form is marked | Histological score |
| Normally | - | 0.65±0.38 | 0.74±0.41 |
| Model | - | 5.85±1.07 ▼ | 5.18±0.90 ▼ |
| SASP | 0.3 | 2.43±0.68* | 1.96±0.72* |
| BJC is high | 16.0 | 2.83±0.88* | 2.37±0.81* |
| In BJC | 8.0 | 3.29±0.84* | 3.37±0.76* |
| BJC is low | 4.0 | 3.93±0.87* | 3.85±0.92* |
Compare with normal group,
▼p<0.05; Compare with model group, * P<0.01; N=10.
The impact of the active and MDA level of ulcerative colitis in rats colon SOD, MPO that BJC induces TNBS: the results are shown in Table 2.Model group rats colonic mucosal tissue SOD value is significantly lower than normal group, and MPO, MDA value is significantly higher than normal group (P<0.05).The each dosage group of BJC and SPSA group all can raise ulcerative colitis in rats SOD value, reduce MPO, MDA value (P<0.05).
Table 2BJC on the impact of SOD, MPO and MDA in ulcerative colitis in rats colon (
)
Compare with normal group,
▼p<0.05; Compare with model group, * P<0.01; N=10.
Herba Patriniae Changning tablet is to the intervention effect to ulcerative colitis in rats colon IL-1 β, IL-10 and TNF-alpha expression
Concrete steps are as follows:
Material and instrument: Herba Patriniae Changning tablet (BJC); TNBS is produced by Sigma Co., USA, and lot number is 090M5000, purchased from Shanghai Sigma-Aldrich Bioisystech Co., Ltd; Balsalazide sodium sheet, specification: 0.5g/ sheet, Shanxi Ante Biological Pharmaceutical Co., Ltd. produces, the accurate word H20041706 of traditional Chinese medicines; Chloral hydrate is produced by Tianjin Ke Miou chemical reagent company limited, and lot number is 20090922; AR level dehydrated alcohol is produced by Tianjin great Mao chemical reagent company limited, and lot number is 20090318; Rat TNF-α, IL-1, IL-10 enzyme-linked immunologic detecting kit is U.S. company BD product, by upper Hypon bio tech ltd import subpackage.SD rat 40, male and female half and half, body weight 180-220g, provides purchased from Guangdong Medical Lab Animal Center, quality certification SCXK (Guangdong) 2008-0002, and test prospective adaptation is raised one week.Electronic analytical balance (model is FA1004) is Shanghai Precision Scientific Apparatus Co., Ltd; Calorstat (model: HPX-9052MBE) is Shanghai Boxun Industrial Co., Ltd.; RM2135 type cycle type microtome, EG1140 paraffin wax embedding, HI1210 spread out sheet machine, HI1220 dehydrator, Leca company; Automatic clinical chemistry analyzer, Italian ECHO company; LeicaDMR advanced studies type biological microscopy imaging system, German Leica company; Multi-functional microplate reader, perkinElmer company of the U.S.; HC-3618R High speed refrigerated centrifuge, Anhui Zhong Kezhongjia scientific instrument company limited; Disposable sterilized injector, operating theater instruments etc.
Test method:
Pharmaceutical formulations: with mortar by the granule pulverization in Herba Patriniae intestinal Yiganning capsule, adding distil water is made into the solution that drug level is 0.2g/mL.With mortar by balsalazide sodium sheet pulverization, the sodium carboxymethyl cellulose with 0.5% is made into the solution that drug level is 0.1g/mL.
Grouping, modeling and administration: 40 rats are divided into 4 groups at random, often organize 10: normal group, model group, BJC group and balsalazide group, except normal group, all the other are respectively organized Rat Fast and can't help water 24h, after 10% chloral hydrate intraperitoneal injection of anesthesia, slowly 8cm is inserted by anus with rat oral gavage needle tubing, model group and BJC group push modeling solution (using the 100mg/kgTNBS that 50% ethanol 0.25mL is made into), normal group gives same volume normal saline enema, after each group of rat handstand 30s, lie on the back and put back to cage.After coloclysis 24h, normal group and model group gavage normal saline 10mL/kg.d, BJC group gavage Herba Patriniae aqueous solution 10mL/kg.d, balsalazide group gavage balsalazide sodium aqueous solution 10mL/kg.d (i.e. balsalazide sodium 1g/kg, rat dosage is adult's clinical dosage 10 times) gavage, continuous 10d.
Draw materials: each treated animal fasting 24h after last administration completes, cervical dislocation is put to death, and opens abdominal cavity and takes out colon rapidly, cut off enteric cavity, clean intestinal tissue with normal saline along mesentery edge, be laid on ice pan, perusal colon general appearance form.Get perusal colonic pathological change the most seriously to locate, after a part is put and is fixed in 10% neutral formalin, paraffin embedding, section, HE dyes, tissues observed Histopathologic changes under light microscopic, remainder is placed on-80 DEG C, for the detection of TNF-α, IL-1 β, IL-10 level.
Ordinary circumstance is observed: record every day such as rat body weight, stool, food-intake, fur change and active situation etc.Relatively rat body weight change (when comparatively experiment starts changes of body mass percentage ratio), stool and hemafecia situation before and after administration, carry out DAI scoring.
The damage of colon general form is observed: observe colon hyperemia, ulcer and intestinal wall and thicken situation, mark to the damage of colon general form.
Colon's pathological examination: light Microscopic observation colon pathological change, marks to colon's pathology.
Colon TNF-α, IL-1 β, IL-10 horizontal detection: every rat takes colon 200mg, put into test tube, add the homogenate of 9 times of volumes ice cold PBS buffer (PH7.4) ice baths again, thawing 2 times, refrigerated centrifuge 4 DEG C of centrifugal 10min of 3000r/min, get supernatant, adopt double antibody sandwich ELISA to detect the level of supernatant TNF-α, IL-1 β, IL-10.
Statistical procedures: the data obtained all represents with mean ± standard deviation (± s).Statistical procedures software spss13.0 is adopted to carry out statistical analysis, between multisample, mean compares employing one factor analysis of variance (one-wayANOVA), and group difference compares with LSD-t (variance is mutually neat), Tamhane ' sT2 (heterogeneity of variance).
Result of the test:
Ordinary circumstance: normal rats situation is all normal, is quick on the draw, hair luster, and diet is normal, without suffering from diarrhoea and having blood in stool.Model group rats, lethargy, few dynamic, hair is matt in a jumble, and dietary amount reduces, and occurs diarrhoea in various degree and has blood in stool.Rat spirit after pharmaceutical intervention takes a turn for the better gradually, and dietary amount, weight recover gradually, and most of rat stool is gradually shaped, partial rat is rotten just continue to execution before.Put to death before all modeling group rat weights significantly lower than normal group (P<0.01), model group lower than other modeling groups, each modeling group difference not statistically significant.Modeling group rat DAI marks apparently higher than normal group (P<0.05), model group is apparently higher than other modeling groups (P<0.05), but two treatment group no significant difference (P>0.05) (table 3).
Table 3 respectively group rat weight, DAI marks (n=10, mean ± SD)
| Group | Weight (g) | DAI marks |
| Normally | 295.00±6.73 | 0 |
| Model | 210.00±18.16 ▼ | 3.06±1.35 ◆ |
| BJC | 234.00±29.07 ▼ | 0.84±0.91 ◆# |
| Balsalazide | 248.50±26.52 ▼ | 0.92±1.06 ◆# |
Compare with normal group,
▼p<0.05,
◆p<0.01; Compare with model group,
#p<0.05; N=10.
Colon general form Injury score: normal rats colon general form is normal, pleat clean mark, intestinal wall is without thickening and adhesion, and mucosa has no congested, rotten to the corn and ulcer.The visible intestinal adhesions of model group rats, intestinal tube pneumatosis stercoroma, intestinal wall thickens, and gauffer disappears, the extensive congestion and edema of intestinal mucosa, and visible significantly ulcer stove.After pharmaceutical intervention, BJC group, balsalazide group rat colon pathological changes obviously alleviate, and intestinal wall is without thickening, and gauffer is normal, and intestinal mucosa is without obvious rotten to the corn, ulcer, and only hyperemia, edema are seen in local.All modeling group rat colon general form scorings are significantly higher than normal group (P<0.01), BJC group, balsalazide group comparatively model group significantly decline (P<0.01), but no significant difference (table 4) between two treatment groups.
Table 4 is the damage of group rat colon general form and histopathological scores (n=10, mean ± SD) respectively
| Group | Weight (g) | DAI marks |
| Normally | 0 | 0 |
| Model | 2.86±0.79 ◆ | 5.97±1.71 ◆ |
| BJC | 1.15±0.93 ◆* | 2.14±1.62 ◆* |
| Balsalazide | 1.22±0.87 ◆* | 2.08±1.55 ◆* |
Compare with normal group,
◆p<0.01; Compare with model group,
*p<0.01; N=10.
Colon's histological scores: under normal rats colonoscope, tissues observed form is normal, and each Rotating fields of colon wall is clear, has no edema, erosion and ulcer, a small amount of lymphocytic infiltration of interstitial.The visible mucous layer of model group rats colon, the extensive ulcer of Submucosa, neutrophilic granulocyte and lymphocytic infiltration, body of gland destroys, structure disturbance, and goblet cell reduces, crypts structural distortion, and crypts inflammation and abscess are formed.After pharmaceutical intervention, BJC group, balsalazide group rat colon organize lesions visible obviously more sick than model group light, and ulcer healing, crypt abscess disappears, and congestion and edema obviously alleviates, and inflammatory cell infiltration obviously reduces.All modeling group rat colon histopathological scores are significantly higher than normal group (P<0.01), BJC group, balsalazide group comparatively model group significantly decline (P<0.01), but no significant difference (table 4) between two treatment groups.
Colon tissue tumor necrosis factor TNF-alpha, IL-1, IL-10 level: compared with normal group, model group rats colon proinflammatory cytokine TNF-α, IL-1, IL-10 level significantly raises, there is pole significant (P<0.01), but the no significant difference between two treatment groups.Compared with model group, two treatment group rat colon tissue T NF-α, IL-1, IL-10 levels significantly decline (P<0.01).No significant difference (table 3) between two treatment groups.Compared with normal group, colon's IL-10 level of model group, BJC treated animal significantly raises (P<0.01), balsalazide group not statistically significant.Compare with model group, balsalazide group rat colon organizes IL-10 level significantly to decline (P<0.01), BJC group not statistically significant.BJC group rat colon organizes IL-10 level to be significantly higher than balsalazide group (P<0.01) (table 5).
Table 5 is group rat colon histiocytokine level (n=10, mean ± SD) respectively
| Group | TNF-α | IL-1 | IL-10 |
| Normally | 460.65±102.14 | 63.18±15.04 | 230.18±31.82 |
| Model | 750.64±215.22 ◆ | 134.06±28.73 ◆ | 358.25±66.47 ◆ |
| BJC | 498.28±138.41 * | 59.27±15.47 * | 256.41±48.09 ◆ |
| Balsalazide | 455.63±142.09 * | 54.17±12.23 * | 228.77±52.01 * |
Compare with normal group,
◆p<0.01; Compare with model group,
*p<0.01; N=10.
Claims (10)
1. a preparation method for Herba Patriniae extract, comprises the steps:
1) Herba Patriniae is pulverized, add flooding, lixiviating solution is concentrated, obtain concentrated solution;
2) concentrated solution water-saturated n-butanol is extracted, extract is merged, remove solvent, obtain residue;
3) by residue water dissolution, solution polyamide is adsorbed, washes with water, collect water lotion, dry, obtain dry powder;
4) dry powder is dissolved, by solution absorption with macroporous adsorbent resin, wash with water, abandon water lotion, then with 30 ~ 95% ethanol elutions, collect ethanol elution, namely concentrate drying obtains Herba Patriniae extract.
2. preparation method according to claim 1, it is characterized in that: the preparation manipulation of concentrated solution is: pulverized by Herba Patriniae, add the water soaking of medical material volume 2 ~ 5 times amount, soak time 0.5 ~ 2h, use the water extraction 1 ~ 3 time of quality of medicinal material 6 ~ 14 times amount again, extract 0.5 ~ 2 hour, merge extractive liquid, at every turn, filter, filtrate is condensed into the concentrated solution that concentration is the former medicine/mL of 0.5 ~ 1g.
3. preparation method according to claim 1 and 2, is characterized in that: the preparation manipulation of residue is: the water-saturated n-butanol getting concentrated solution 1 ~ 3 times of volume extracts 1 ~ 3 time repeatedly, merges butanol extraction liquid, and recycling design is to dry.
4. preparation method according to claim 1, is characterized in that: the amount ratio of residue and polyamide is: the former medicine of 1g/(1 ~ 3) g polyamide.
5. preparation method according to claim 1, is characterized in that: the amount ratio of dry powder and macroporous adsorbent resin is: the former medicine of 1g/(1 ~ 3) g macroporous adsorbent resin.
6. preparation method according to claim 1, is characterized in that: the model of macroporous adsorbent resin is the one in D101, D140, D141, AB-8.
7. preparation method according to claim 1, is characterized in that: comprise the steps:
1) Herba Patriniae is pulverized, add the water soaking of medical material volume 2 ~ 3 times amount, soak time 0.5 ~ 2h, use the water extraction 1 ~ 2 time of quality of medicinal material 8 ~ 12 times amount again, extract 1 ~ 1.5 hour, merge extractive liquid, at every turn, filter, filtrate is condensed into the concentrated solution that concentration is 0.6 ~ 0.9 former medicine g/mL;
2) water-saturated n-butanol getting concentrated solution 1 ~ 2 times of volume extracts 1 ~ 3 time repeatedly, merges butanol extraction liquid, and recycling design, to dry, obtains residue;
3) get the solution that residue water dissolution becomes the former medicine/mL of 0.6 ~ 0.9g, solution polyamide is adsorbed, washes with water, collect water lotion, dry, obtain dry powder;
4) dry powder is dissolved into the solution of the former medicine/mL of 0.6 ~ 0.9g, by solution absorption with macroporous adsorbent resin, washes with water, abandon water lotion, then with 55 ~ 75v/v% ethanol elution, collect ethanol elution, namely concentrate drying obtains Herba Patriniae extract.
8. treat a compositions for ulcerative colitis, its active component is Herba Patriniae extract, it is characterized in that: the preparation method of Herba Patriniae extract is as described in claim 1 ~ 7 any one.
9. compositions according to claim 8, is characterized in that: the dosage form of compositions is oral formulations.
10. compositions according to claim 9, is characterized in that: oral formulations is selected from tablet, capsule, granule, drop pill.
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| CN201510833390.5A CN105326872A (en) | 2015-11-25 | 2015-11-25 | Preparation method for herba patriniae extract with therapeutical effect on ulcerative colitis |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110302282A (en) * | 2019-06-06 | 2019-10-08 | 中国人民解放军南部战区总医院 | Herba Patriniae extract is preparing the application in overriding resistance Acinetobacter bauamnnii drug |
| CN111920875A (en) * | 2020-09-08 | 2020-11-13 | 江西中医药大学 | Patrinia extract with anti-atherosclerosis effect |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101168571A (en) * | 2007-11-28 | 2008-04-30 | 李鑫 | Method for exactly extracting antiviral field pennycress polysaccharide from field pennycress |
-
2015
- 2015-11-25 CN CN201510833390.5A patent/CN105326872A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101168571A (en) * | 2007-11-28 | 2008-04-30 | 李鑫 | Method for exactly extracting antiviral field pennycress polysaccharide from field pennycress |
Non-Patent Citations (3)
| Title |
|---|
| 常文贵等: "黄花败酱黄酮类化合物的提取与分析", 《生物学杂志》 * |
| 张永强: "败酱草总皂甙抗小鼠宫颈癌活性研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 韩亮等: "败酱草提取物对三硝基苯磺酸诱导的大鼠结肠炎的保护作用", 《广东药学院学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110302282A (en) * | 2019-06-06 | 2019-10-08 | 中国人民解放军南部战区总医院 | Herba Patriniae extract is preparing the application in overriding resistance Acinetobacter bauamnnii drug |
| CN111920875A (en) * | 2020-09-08 | 2020-11-13 | 江西中医药大学 | Patrinia extract with anti-atherosclerosis effect |
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Application publication date: 20160217 |