CN102973554A - Medical application of alantolactone - Google Patents
Medical application of alantolactone Download PDFInfo
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- CN102973554A CN102973554A CN2012105534379A CN201210553437A CN102973554A CN 102973554 A CN102973554 A CN 102973554A CN 2012105534379 A CN2012105534379 A CN 2012105534379A CN 201210553437 A CN201210553437 A CN 201210553437A CN 102973554 A CN102973554 A CN 102973554A
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Abstract
The invention relates to the field of medicine, and in particular relates to medical application of alantolactone. The alantolactone can be used for preventing and treating enteritis.
Description
Technical field
The present invention relates to the Medicines and Health Product field, particularly relate to a kind of medical usage of alantolactone.
Background technology
Ulcerative colitis (Ulcerative colitis, UC) is the inflammatory bowel chronic, that repeatedly show effect that betides colon.Clinical main manifestations is diarrhoea, mucus bloody purulent stool, stomachache.Major complications has: hemorrhage, intestinal perforation, anal fistula, intestinal obstruction, colon cancer and septic shock etc.
Alantolactone (Alantolactone) is a kind of sesquiterpene lactones, is one of main active of Radix Inulae, also is one of main effective ingredient of four-Tibet inula root soft capsule.Alantolactone has anthelmintic, the multiple pharmacologically active such as antibiotic.
Summary of the invention
Purpose of the present invention aims to provide a kind of new medical usage of alantolactone.
Specifically, the invention provides a kind of alantolactone and prevent and treat the medicine of enteritis or the application in the food in preparation.
In a preference, described enteritis is ulcerative colitis.
The details of various aspects of the present invention will be able to detailed description in chapters and sections subsequently.By hereinafter and the description of claim, characteristics of the present invention, purpose and advantage will be more obvious.
Description of drawings
Fig. 1 has embodied the impact of alantolactone on the ulcerative colitis Mouse Weight;
Fig. 2 has embodied the impact of alantolactone on ulcerative colitis mice hemafecia incidence rate;
Fig. 3 has embodied the impact of alantolactone on ulcerative colitis mice colon length;
Fig. 4-5 has embodied the impact of alantolactone on ulcerative colitis mice colon pathology;
Fig. 6-7 has embodied alantolactone to the TNF-α of ulcerative colitis mice colon and the impact of IL-6mRNA expression;
(in above-mentioned figure: a represents Normal group, and b representation model matched group, c represent the SASP group, and d represents the alantolactone group)
The specific embodiment
The part of coming out of the present invention is based on so unexpected discovery: alantolactone can obviously improve the losing weight of ulcerative colitis mice, hemafecia, colon shortens and the symptom such as histopathology damage, and can reduce the content of the inflammatory cytokine TNF-α of colon and IL-6.Therefore, alantolactone can be used for preparation and prevents and treats medicine or the health food of enteritis, particularly ulcerative colitis.
And then, the invention provides alantolactone and prevent and treat the medicine of enteritis or the application in the health food in preparation.
The molecular formula of alantolactone of the present invention is: C
15H
20O
2, molecular weight is: 232.31, and its structural formula is as follows:
Alantolactone of the present invention can be bought from Sigma chemical company, Chengdu Man Site bio tech ltd etc. by commercial sources and obtain.Its purity all meets medicinal standard.The purity of alantolactone of the present invention with 98% the best.
Alantolactone of the present invention can use separately or use with the form of pharmaceutical composition.Pharmaceutical composition comprises alantolactone of the present invention and the pharmaceutically suitable carrier as active component.Preferably, pharmaceutical composition of the present invention contains the alantolactone of the present invention as active component of 0.1-99.9% percentage by weight." pharmaceutically suitable carrier " can not destroy the pharmaceutical active of alantolactone of the present invention, its effective dose simultaneously, and can bringing into play pharmaceutical carrier, to make the consumption of time spent nontoxic to human body.
Described pharmaceutically suitable carrier includes but not limited to: soft phospholipid, aluminium stearate, aluminium oxide, ion exchange material, self-emulsifying drug delivery system, tween or other surfactants, serum albumin, buffer substance such as phosphate, glycine, sorbic acid, water, salt, electrolyte such as sulfate protamine, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, magnesium silicate, satisfied fatty acid partial glycerol ester admixture etc.
The excipient substance that other are commonly used such as binding agent (such as microcrystalline Cellulose), filler (such as starch, glucose, Lactis Anhydrous and lactose beadlet), disintegrating agent (such as cross-linked pvp, crosslinked carboxymethyl fecula sodium, cross-linking sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose), lubricant (such as magnesium stearate) and absorption enhancer, absorption carrier, flavouring agent, sweeting agent, excipient, diluent, wetting agent etc.
Alantolactone of the present invention with and pharmaceutical composition can and can pass through intestinal or non-intestinal or topical routes by this area conventional method preparation.Oral formulations comprises capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum etc.; Non-intestinal drug delivery agent comprises injection etc.; Local administration preparation comprises cream, patch, ointment, spray etc.Be preferably oral formulations.
Alantolactone of the present invention take and the route of administration of pharmaceutical composition can be as oral, Sublingual, percutaneous, through muscle or subcutaneous, mucocutaneous, vein, urethra, vagina etc.
Except making medicament, also can in alantolactone of the present invention, add the various food additive such as antioxidant, pigment, enzyme preparation, make health food by the conventional method of this area.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment only to be used for explanation the present invention and be not used in and limit the scope of the invention.The experimental technique of unreceipted actual conditions in the following example is usually according to normal condition or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise all percent, ratio, ratio or umber by weight.
Unless otherwise defined, the same meaning that employed all specialties and scientific words and one skilled in the art are familiar with in the literary composition.In addition, any method similar or impartial to described content and material all can be applicable in the inventive method.The usefulness that better implementation method described in the literary composition and material only present a demonstration.
The above-mentioned feature that the present invention mentions, or the feature that embodiment mentions can combination in any.All features that patent specification discloses can with any composition forms and usefulness, each feature that discloses in the description can anyly provide the alternative characteristics of identical, impartial or similar purpose to replace.Therefore except special instruction is arranged, the feature that discloses only is the general example of equalization or similar features.
Embodiment 1:
1, experiment material
1.1 medical material
Alantolactone (CAS:546-43-0, MW 232.31, HPLC purity 〉=98%, Chengdu Man Site bio tech ltd); Dextran sulfate sodium (MW 36000-50000 is produced by U.S. MP Biomedicals company for DSS, CAS:9011-18-1); Sulfasalazine (SASP, CAS:599-79-1, molecular weight 398.39, HPLC purity 〉=98% is produced by U.S. Sigma-Aldrich company).
1.2 laboratory animal
The C57BL/6 female mice of body weight 20 ± 2g (8 week) is provided by Shanghai Univ. of Traditional Chinese Medicine's Experimental Animal Center, the animal quality certification number: SYXK(Shanghai) 2009-0069.Place conventional feeding environment, ad lib and drinking-water.
2, experimental technique
2.1 the foundation of Ulcerative Colitis Model
The C57BL/6 female mice of selective body weight average one (20 ± 2g), adopt international ulcerative colitis modeling method (Gastroenterology, 1990,98:694-702; DigestiveDisease and Science, 1993,38:1722-34; And U.S. Patent No. 5,869,048), in the research incipient stage, make mice ad lib and drinking-water, after the adaptive phase, change water into 5%(w/v in the early stage) DSS, freely drank 5 days, to cause ulcerative colitis.
2.2 grouping and medication
Normal group: 15, give normal diet.
Model control group: 15, give 5%DSS solution and freely drank 5 days, changed normal drinking water into since the 5th day, gavage gave continuous normal saline 7 days simultaneously.
The SASP group: 15, give 5%DSS solution and freely drank 5 days, changed normal drinking water into since the 5th day, gavage gives SASP(350mg/kg simultaneously) continuous 7 days.
The alantolactone group: 15, give 5%DSS solution and freely drank 5 days, changed normal drinking water into since the 5th day, gavage gave alantolactone (50mg/kg) continuous 7 days simultaneously.
Experimental session, every day, fresh preparation dosage particles with the dissolving of 0.5% sodium carboxymethyl cellulose, and used in 1 hour in preparation.
2.3 the assessment of enteritis
Diarrhoea, hemafecia, histopathological analysis etc. are all with reference to early-stage Study (PLoS One.2012,7:e36075) the described method that publishes thesis.Experimental session records body weight change, diarrhoea and hemafecia symptom every day.Mice was put to death in the dislocation of anesthesia and cervical vertebra after experiment finished, and opened the abdominal cavity, took out colon, measured the colon length from the caecum to the rectum.And get DC and make tissue slice, carry out H﹠amp; E dyeing is to H﹠amp; E dyeing colon specimen is carried out pathological score: (1) inflammatory cell ooze out standards of grading: the minute quantity inflammatory cell is arranged in the 0 minute-mucosa lamina propria, there is the inflammatory cell in more inflammatory cell or the mucosa lamina propria to increase in the 1 minute-mucosa lamina propria, 2 minutes-inflammatory cell diffuses to Submucosa, and 3 minutes-holostrome all has inflammatory cell to ooze out; (2) tissue damage standards of grading: 0 minute-without mucosa injury, 1 minute-discrete mucosal epithelium was damaged, 2 minutes-top layer mucosal erosion, 3 minutes-mucosa is damaged and expand to the intestinal wall deep layer widely.Oozing out of inflammatory cell scored and tissue damage score addition, calculate histopathology scoring (1-6 divides).
2.4 fluorescence quantitative RT-RCR detects the TNF-α of colon and the mrna expression of IL-6
Effect experiment is got colon after finishing in the body, extracts the total RNA of colon fragment also quantitatively with Trizol reagent, gets 3 μ g RNA and prepares cDNA with the M-MLV reverse transcription.Take cDNA as template, increase at the ABI7300 quantitative real time PCR Instrument.PCR reaction system: 2 * SYBR Premix Ex Taq(TakaRa, cat:DRR420A) 5 μ L, Forward Primer 0.4 μ L, Reverse Primer0.4 μ L, 50 * ROX Reference Dye, 0.4 μ L, cDNA 1 μ L, dH
2O 2.8 μ L, cumulative volume are 10 μ L.PCR response parameter: 95 ℃, 30s denaturation, 95 ℃, 5s degeneration, 60 ℃, 30s annealing, totally 40 circulations.Each amplification arranges β-actin internal reference.Primer sequence is as follows:
MTNF-α: positive-sense strand 5 '-CGTGGAACTGGCAGAAGAGG-3 ',
Antisense strand 5 '-AGACAGAAGAGCGTGGTGGC-3 ';
MIL-6: positive-sense strand 5 '-ACCACGGCCTTCCCTACTTC-3 ',
Antisense strand 5 '-CATTTCCACGATTTCCCAGA-3 ';
β-Actin: positive-sense strand 5 '-CAGCCTTCCTTCTTGGGTAT-3 ',
Antisense strand 5 '-GGTCTTTACGGATGTCAACG-3 '.
Carry software with the pcr amplification instrument and carry out quantitative fluorescence analysis, and draw the Ct value.Draw and respectively organize gene with respect to the expression of β-Actin by calculating 2-△ Δ CT.
2.5ELISA method detects the content of the TNF-α of colon and IL-6
Mice was put to death in the dislocation of anesthesia and cervical vertebra after experiment finished, and opened the abdominal cavity, took out colon, the rear homogenate of weighing.Homogenate is in 4 ℃, turns 2000/centrifugal 10 minutes of min.Get 10 μ L supernatant, with ELISA test kit (R﹠amp; D systems, Minneapolis, MN, USA) measure the content of TNF-α and IL-6, the equal reference reagent box of concrete operations description.Measure protein content with the BCA method.The result is expressed as pg/mg protein.
2.6 statistical analysis
All experimental datas all repeat 3 times, and the result represents with mean+SD, adopt the SPSS16.0 statistical software that experimental data is adopted one way analysis of variance (One-way ANOVA) and LSD check, and there is significance P＜0.05 for difference statistically.
3, result
3.1 alantolactone is to the therapeutical effect of ulcerative colitis
3.1.1 the impact on the ulcerative colitis Mouse Weight
Fig. 1 has embodied the impact of alantolactone on the ulcerative colitis Mouse Weight.(amount to 12 days, comprise 5 days of processing with DSS and 7 days of treated with medicaments subsequently) in whole experimentation, the body weight change of Normal group mice is steady, and (19.8 ± 0.2g) have increased by 1.4% to the 12nd day body weight; The body weight continuous decrease of model group mice, (17.3 ± 0.2g) to alleviate rate be 11.3% to the 12nd day body weight; SASP group (18.4 ± 0.3g) and alantolactone group (the body weight continuous decrease to the of 18.6 ± 0.2g) mices 7 days, fall is less than model group, tend to be steady subsequently and slightly raise, the 12nd day the rate of losing weight is respectively 5.5% and 4.4%, and comparing with model group all has significant difference (P＜0.01 and P＜0.001).Illustrate that SASP and alantolactone all can obviously improve the symptom that loses weight of the ulcerative colitis mice that DSS causes.
3.1.2 the impact on ulcerative colitis mice hemafecia incidence rate
Fig. 2 has embodied the impact of alantolactone on ulcerative colitis mice hemafecia incidence rate.(amount to 12 days, comprise 5 days of processing with DSS and 7 days of treated with medicaments subsequently) in whole experimentation, mice occurs without hemafecia Normal group.Since the 4th day, 3 groups of mices of all the other except Normal group begin to occur the hemafecia symptom, and were the most serious with the model group mice.The model group hemafecia continues to occur to the 10th day, tends to be steady subsequently, and the 12nd day hemafecia incidence rate is 96.7%; SASP group and alantolactone group mice hemafecia continue to occur to the 6th day, tend to be steady subsequently and decline is slightly arranged, and the 12nd day hemafecia incidence rate is respectively 46.7% and 33.3%, and comparing with model group all has significant difference (P＜0.001).Illustrate that SASP and alantolactone all can obviously improve the hemafecia symptom of the ulcerative colitis mice that DSS causes.
3.1.3 the impact on ulcerative colitis mice colon length
Fig. 3 has embodied the impact of alantolactone on ulcerative colitis mice colon length.(8.8 ± 0.3cm) compare, and (colon of 5.5 ± 0.5cm) mices obviously shortens model group, and LVFS is 37.7% with Normal group; SASP group (6.8 ± 0.5cm) with the alantolactone group (the colon LVFS of 7.3 ± 0.4cm) mices is respectively 22.6% and 17.7%, and comparing with model group all has significant difference (P＜0.05 and P＜0.01).Illustrate that the colon that SASP and alantolactone all can obviously improve the ulcerative colitis mice that DSS causes shortens symptom.
3.1.4 the impact on ulcerative colitis mice colon pathology
Fig. 4 has embodied the injury repairing effect of alantolactone to ulcerative colitis mice colon.The colon of model group mice is typical inflammation mucosa performance, visible mucosa, Submucosa even flesh layer massive inflammatory cells infiltrated, disorderly, the fracture of organizational structure, hemorrhage, the edema of Submucosa, telangiectasis; The visible pathological changes of colon of SASP group mice and alantolactone group mice alleviates, ulcer healing, and Submucosa is hemorrhage to be alleviated.
Fig. 5 has embodied colon specimen pathological score result.According to 1-6 minute standards of grading that represent the inflammatory tissue damage, the score of Normal group mice is 0; Model group mice score (5.5 ± 0.5 minutes) significantly increases; SASP group (2.5 ± 0.5 minutes) and the score of alantolactone group (2.0 ± 0.4 minutes) mice all reduce, and comparing with model group all has significant difference (P＜0.001).
To sum up colon's pathological analysis result shows, SASP and alantolactone all can significantly improve the damage of colonic mucosa histopathology and the inflammatory cell infiltration symptom of the ulcerative colitis mice that DSS causes.
3.1.5 on the TNF-α of ulcerative colitis mice colon and the impact of IL-6 expression
Fig. 6 has embodied the impact of alantolactone on the TNF-alpha expression of ulcerative colitis mice colon.Model group (255.7 ± 10.2) TNF-of mice colon alpha expression is apparently higher than Normal group (1.0 ± 0.1); After gavage gave SASP (89.7 ± 5.5) or alantolactone (41.2 ± 5.8), the TNF-alpha expression of mice colon descended, and the two is compared with model group all significant difference (P＜0.001).Show that SASP and alantolactone all can significantly reduce the expression of TNF-α in the inflammatory reaction that DSS causes.
Fig. 7 has embodied the impact of alantolactone on the IL-6 expression of ulcerative colitis mice colon.Model group (196.4 ± 14.7) IL-6 of mice colon expresses apparently higher than Normal group (1.0 ± 0.56); After gavage gave SASP (64 ± 8.9) or alantolactone (44.9 ± 5.8), the IL-6 of mice colon expressed decline, and the two is compared with model group all significant difference (P＜0.001).Show that SASP and alantolactone all can significantly reduce the expression of IL-6 gene in the inflammatory reaction that DSS causes.
3.1.6 the impact on the TNF-α of ulcerative colitis mice colon and IL-6 content
Get the rear homogenate of weighing of colon specimen, the content with TNF-α and IL-6 in the ELISA kit measurement homogenate supernatant the results are shown in Table 1.
Table 1. alantolactone is on the impact of the TNF-α of ulcerative colitis mice colon and IL-6 content
###P＜0.001, model group is compared with matched group;
* *P＜0.001, treatment group is compared with matched group.
Table 1 has embodied the impact of alantolactone on the TNF-α of ulcerative colitis mice colon and IL-6 content.Compare with Normal group (14.6 ± 1.7), model group (255.4 ± 13.5) TNF-of mice colon alpha content is apparently higher than Normal group (P＜0.001); Gavage gives SASP(84.6 ± 8.5) or alantolactone (53.4 ± 2.9) after, the TNF-alpha content of mice colon descends, and compares with model group, and significant difference (P＜0.001) is all arranged.Show that SASP and alantolactone all can significantly reduce the tissue content of TNF-α in the inflammatory reaction that DSS causes.
In addition, compare with Normal group (28.9 ± 2.3), model group (214.3 ± 17.9) IL-6 of mice colon content apparently higher than Normal group (
* *P＜0.001); After gavage gave SASP (93.3 ± 7.9) or alantolactone (47.6 ± 3.5), the IL-6 content decrease of mice colon was compared with model group, and significant difference (P＜0.001) is all arranged.Show that SASP and alantolactone all can significantly reduce the tissue content of IL-6 in the inflammatory reaction that DSS causes.
To sum up result of study shows, alantolactone can obviously improve the losing weight of ulcerative colitis mice, hemafecia, colon shortens and the symptom such as inflammatory tissue damage, and significantly reduces expression and the tissue content of inflammatory cytokine TNF-α and IL-6.
Many aspects involved in the present invention have been done as above and have been set forth.Yet, it should be understood that before not departing from spirit of the present invention to put that those skilled in the art can be equal to change and modification to it, described change and modification fall into the coverage of the application's claims equally.
Claims (2)
1. alantolactone is prevented and treated the medicine of enteritis or the application in the food in preparation.
2. application according to claim 1 is characterized in that, described enteritis is ulcerative colitis.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104666295A (en) * | 2015-02-04 | 2015-06-03 | 广东省农业科学院动物卫生研究所 | Application of alantolactone in preparation of medicines for resisting cryptosporidium parvum |
| CN104771430A (en) * | 2015-04-10 | 2015-07-15 | 上海交通大学 | Medicine for treating irritable bowel syndrome |
| CN106083785A (en) * | 2015-01-22 | 2016-11-09 | 浙江大学宁波理工学院 | A kind of Preparation method and use of eudesmane type natural product derivant 13 benzylamine alantolactone |
| CN106474109A (en) * | 2016-10-09 | 2017-03-08 | 天承南运(天津)科技有限公司 | The application in preparation treatment inflammatory bowel medicine of isoalantolactone derivant and its salt |
| CN106491593A (en) * | 2016-10-09 | 2017-03-15 | 天承南运(天津)科技有限公司 | The application of alantolactone derivant and its salt in treatment inflammatory bowel medicine is prepared |
| CN107157983A (en) * | 2017-05-19 | 2017-09-15 | 苏州大学附属第医院 | Application of the alantolactone in preventing and treating hepar damnification medicine is prepared |
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| WO2004066912A2 (en) * | 2003-01-31 | 2004-08-12 | Technion Research & Development Foundation Ltd. | Anti-inflammatory compositions and uses thereof |
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2012
- 2012-12-19 CN CN201210553437.9A patent/CN102973554B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2004066912A2 (en) * | 2003-01-31 | 2004-08-12 | Technion Research & Development Foundation Ltd. | Anti-inflammatory compositions and uses thereof |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106083785A (en) * | 2015-01-22 | 2016-11-09 | 浙江大学宁波理工学院 | A kind of Preparation method and use of eudesmane type natural product derivant 13 benzylamine alantolactone |
| CN106083785B (en) * | 2015-01-22 | 2018-03-09 | 浙江大学宁波理工学院 | A kind of Preparation method and use of the benzylamine alantolactone of eudesmane type natural products derivative 13 |
| CN104666295A (en) * | 2015-02-04 | 2015-06-03 | 广东省农业科学院动物卫生研究所 | Application of alantolactone in preparation of medicines for resisting cryptosporidium parvum |
| CN104771430A (en) * | 2015-04-10 | 2015-07-15 | 上海交通大学 | Medicine for treating irritable bowel syndrome |
| CN104771430B (en) * | 2015-04-10 | 2018-05-08 | 上海交通大学 | For treating the medicine of intestinal irritable syndrome |
| CN106474109A (en) * | 2016-10-09 | 2017-03-08 | 天承南运(天津)科技有限公司 | The application in preparation treatment inflammatory bowel medicine of isoalantolactone derivant and its salt |
| CN106491593A (en) * | 2016-10-09 | 2017-03-15 | 天承南运(天津)科技有限公司 | The application of alantolactone derivant and its salt in treatment inflammatory bowel medicine is prepared |
| CN106491593B (en) * | 2016-10-09 | 2019-12-17 | 天承南运(天津)科技有限公司 | Application of alantolactone derivatives and salts thereof in preparation of medicines for treating inflammatory bowel diseases |
| CN107157983A (en) * | 2017-05-19 | 2017-09-15 | 苏州大学附属第医院 | Application of the alantolactone in preventing and treating hepar damnification medicine is prepared |
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