CN101791316A - New application of ginsenoside Rd in preparation of colitis treatment medicament - Google Patents
New application of ginsenoside Rd in preparation of colitis treatment medicament Download PDFInfo
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Abstract
The invention relates to application of ginsenoside Rd in preparation of a colitis treatment medicament, in particular to the application of the ginsenoside Rd in the preparation of an ulcerative colitis treatment medicament. Tests prove the application of the ginsenoside Rd in the preparation of the colitis treatment medicament. The application of the ginsenoside Rd in the preparation of the colitis treatment medicament is that in the preparation of the ulcerative colitis treatment medicament. The application of the ginsenoside Rd in the preparation of the colitis treatment medicament is that in the preparation of nonbacterial chronic colitis treatment medicament. The application of the ginsenoside Rd in the preparation of the colitis treatment medicament is that of ginsenoside Rd oral preparation in the preparation of the ulcerative colitis treatment medicament. By utilizing the characteristics that the ginsenoside Rd is difficult to absorb and mainly enters colon and combining a strong anti-inflammatory effect of the ginsenoside Rd, the application of ginsenoside Rd in preparation of a colitis treatment medicament fully displays the colitis treatment effect of the ginsenoside Rd, discloses the treatment value of an antioxidant for colitis, and has a novel conception.
Description
Technical field:
The present invention relates generally to the purposes of a kind of ginsenoside Rd at preparation treatment colitis medicine.Relate in particular to the purposes of ginsenoside Rd at preparation treatment ulcerative colitis medicine.
Background technology:
(Ulcerative Colitis UC) is all indefinite chronic colitis disease of a kind of cause of disease and pathogenesis to ulcerative colitis, and stomachache, diarrhoea, mucus are its main clinic symptoms with hemafecia just.UC acute fulminant form case fatality rate height, chronic lasting type canceration rate height, easily outbreak repeatedly do not have specific medicament, at present so classified as one of modern difficult treatment by World Health Organization (WHO).Primary disease is higher at American-European countries's sickness rate, and prevalence is 0.4 ‰～1 ‰, does not see accurate statistical report data at home, but is the trend of obvious increase in recent years.Because numerous medical treatment and researcher are to the concern of UC, the new drug of treatment UC constantly occurs.At present, sulfasalazine (SASP) conduct is the medicine commonly used of the treatment ulcerative colitis of widespread usage clinically, curative effect is more obvious, and it and metabolite 5-aminosalicylic acid (5-ASA) thereof are found the effect with stronger removing oxidative stress reaction metabolite, but SASP may produce the anorexia relevant with dosage in some patient treatment processes, feel sick, erythra and hematotoxicity (comprise that mainly neutrophilic granulocyte reduces, agranulocytosis, hemolytic anemia, thrombocytopenia, two kinds of cytopenia, pancytopenia or aplastic anemia) etc. untoward reaction, part male patient motility of sperm may occur and descend and cause infertility.Above-mentioned untoward reaction makes the SASP medication have certain limitation.Therefore find the new drug of treatment ulcerative colitis new, that side effect is low, be still at present an emphasis of new drug research both at home and abroad.
Radix Notoginseng (Panax notoginseng) belongs to the araliaceae ginseng plant, and its main active is a dammarane type four-ring triterpenoid saponin, and is similar to constituent of ginseng, and medicinal part mostly is the root piece.Chinese medicine is theoretical to confirm that Radix Notoginseng has strong tonify deficiency and hemostatic effect, is used for the treatment of some cardiovascular disease, inflammation and various physical distress more, injures the vivo and vitro bleeding that wound causes outward.A large amount of phytochemistries and pharmaceutical research confirm that dammarane type four-ring triterpenoid saponin is its main pharmacological component, wherein the ginsenoside Rd accounts for 4.07% of total saponins, is one of four kinds of saponin that wherein mainly have remarkable pharmacological component (Rg1, Rb1, R1, Rd).The ginsenoside Rd is one of diol type ginsenoside main metabolites in human body intestinal canal, has wide biological activity, and is unique to effects such as cardiovascular and cerebrovascular vessel, nervous system, immune systems, and effect is remarkable aspect analgesia, neuroprotective.The ginsenoside Rd studies the clinical III phase of the protective effect of cerebral ischemia and finished already, is applying for new drug production certification.Our previous work also shows, Rd suppresses the inductive human T-cell's propagation of mitogen ConA, suppresses delayed hypersensitivity, suppress the inductive rat arthritis of glue fork dish glue, extremely strong antiinflammatory action is arranged, and (the research result has been applied for the patent of invention of Rd treatment of arthritis etc. by the inventor, the patent first trial is passed through, see " panaxoside-Rd aqueous solution of propylene glycol preparation and antiinflammatory thereof, immunosuppressant and anti-organ-graft refection's new purposes ", the number of accepting: 200810208119.2).But the medicine (as the aspirin class) with antiinflammatory action might not have the effect of treatment ulcerative colitis, and does not appear in the newspapers the experiment confirm of still needing both at home and abroad about the research that the ginsenoside Rd treats the rat ulcer colitis.
Ginsenoside-Rd is a kind of monomeric compound that gets through the multistep separation from the Chinese medicine panax araliaceae plant, belongs to protopanoxadiol type saponin.Its chinesization formal name used at school is 20-(S)-protopanoxadiol-3[O-β-D-Glucopyranose. (1 → 2)-β-D-pyranglucoside]-20-O-β-D-pyranglucoside, English chemistry 20 (S)-Protopanaxadiol 3-O-[β-D-glucopyranosyl (1 → 2)-β-D-glucopyranosyl by name]-20-O-β-glucopyranoside, chemical constitution is as follows:
C48H82O18·3H2O；1001.20
About from pseudo-ginseng, extracting ginsenoside Rd's method, at first extract thick total saponins, then by macroporous adsorbent resin ethanol gradient elution branch leave away great majority other components, obtain the less Rd component of impurity, again through silicagel column with " chloroform 2 methanol 2 water " solvent gradient elution, the disposable purity that obtains reaches 93.16% Rd monomer.Final purity can reach more than 95%.(Chinese crude drug the 29th is rolled up in March, 2006 the 3rd phase, " rapid batch separation and Extraction ginsenoside Rd's method from pseudo-ginseng " Liu Deyu, the pharmaceutical college of Zhongshan University that once soughed, Guangzhou, Guangdong 510080)
Summary of the invention:
The objective of the invention is to avoid the deficiencies in the prior art part and a kind of ginsenoside Rd is provided the application at preparation treatment colitis medicine.
Purpose of the present invention is by the application of test explanation ginsenoside Rd at preparation treatment colitis medicine.
Described ginsenoside Rd is the application of ginsenoside Rd at preparation treatment ulcerative colitis medicine in the application of preparation treatment ulcerative colitis medicine.
Described ginsenoside Rd is the application of ginsenoside Rd at preparation treatment non-bacterial chronic colitis medicine in the application of preparation treatment colitis medicine.
Described ginsenoside Rd is the application of ginsenoside Rd's oral formulations at preparation treatment ulcerative colitis medicine in the application of preparation treatment colitis medicine.
Described ginsenoside Rd is characterized in that in the application of preparation treatment colitis medicine described oral formulations is for being capsule, microcapsule, liposome, granule, tablet, oral liquid.
Described ginsenoside Rd is in the application of preparation treatment colitis medicine, and the ginsenoside Rd is the ginsenoside Rd's monomeric compound that extracts from Radix Notoginseng, and wherein ginsenoside Rd's monomer is more than 95%; Also comprise ginsenoside Rd's monomeric compound that extraction separation obtains root from araliaceae ginseng plant's Radix Ginseng, Radix Panacis Quinquefolii, stem, leaf, flower, the fruit.
Beneficial effect of the present invention:
(1) confirmed the therapeutical effect of ginsenoside Rd by experiment first to experimental colitis.
(2) ginsenoside and active metabolite thereof rat oral administration artifact utilize low, absorb few, but have the advantages that to accumulate in the colon position in a large number, in conjunction with its powerful antiinflammatory action that has, fully showed the therapeutical effect of ginsenoside Rd to colitis, disclosed the therapeutic value of the calcium ion channel blocker of the calcium pond manipulation with antioxidation to colitis, conception is novel.
(3) adopt form to combine with biochemical measurement (MPO determination of activity), qualitative, quantitative observation medicine makes the evaluation of curative effect of medication more reliable to the influence that ulcerative colitis heals.Mechanism research index the known of colitis of combining closely about pathophysiological mechanism and ginsenoside Rd's anti-oxidation characteristics, change (SOD, GSH-Px), lipid peroxide (MDA) generation as cell infiltration (MPO), oxidative damage and oxidation resistance, deeply, system, disclose treatment of ginsenoside Rd's antioxidation and the relation that promotes the ulcerative colitis healing in all directions, and then illustrate the regulating action of ginsenoside Rd in anti-damage of intestinal mucosa and repair process, the thinking uniqueness, novel.
(4) at present the immunology pathogenesis of UC particularly the cytokine pathogenesis more and more be subject to people's attention.And pro-inflammatory cytokine and press down the important pathogenesis that dysimmunity due to the dysequilibrium between the inflammatory cytokine is regarded as UC, this experiment has confirmed that also the ginsenoside Rd can significantly suppress the generation of short inflammatory cell because of IL-1 β, TNF-α, and the expression that promotes anti-inflammatory cytokines IL-10, set forth from the immunology angle that the ginsenoside Rd treats colitis and it can suppress the expression of colon's pro-inflammatory cytokine, it is closely related to promote to press down scorching factor generation and adjusting immunologic balance, its viewpoint novelty.
(5) the current research data shows, the ginsenoside Rd is a kind of novel current potential independent form membrane receptor calcium activate channel (ROCC) and calcium channel (SOC) blocker of calcium pond manipulation, can suppress receptor-operated Ca significantly by ROCC and SOC approach
2+Interior stream.Existing anti-inflammatory agent such as aspirin class, glucocorticoids and and anti-UC medicine sulfasalazine all do not have a calcium antagonism yet.Regulate inflammatory reaction with specific SOC blocker a new target spot will be provided for the control of inflammation, in conjunction with this subject study result, we predict that this novel non-potential dependent calcium ion channel blocker of ginsenoside Rd may become the novel medicine of treatment ulcerative colitis and other inflammation, and this is provided a kind of new thinking by the treatment that World Health Organization (WHO) classifies modern refractory disease as for colitis.
(6) by system, study the therapeutical effect mechanism of ginsenoside Rd to experimental colitis comprehensively, the omnibearing therapeutical effect mechanism of ginsenoside Rd to colitis that disclosed is for the Drug therapy of ulcerative colitis provides new argument and viewpoint.
The specific embodiment:
Be described in further detail below in conjunction with most preferred embodiment:
Embodiment 1: the ginsenoside Rd treats the dose-effect relationship and the mechanism of action of rat acute ulcerative colitis
1. materials and methods
1.1 medicine and animal Wistar rat, body weight 180～200g,
Available from Lanzhou University's Experimental Animal Center, the animal quality certification number: moving word SCXK (sweet) 2009-0004 of doctor.Ginsenoside Rd's (purity: 〉=95%, the preparation method reference is publication: " chemical compound (1), its extracting method and comprise the pharmaceutical composition of described chemical compound ", Granted publication number: CN1176101C): GuangDong TaiHe Biology Pharmacy Co., Ltd's lot number: 080312; The good magnificent company limited product of Hisense's friendship on the sulfasalazine, lot number: 081013.
1.2 main agents 2,4,6-trinitro-benzene-sulfonic acid (2,4,6-Trinitrobenzenesulfonic acidsolution TNBS) purchases the company in Sigma, 5% (W/V) aqueous solution, article No. P2297-10ML, lot number 20070315); Dehydrated alcohol (rich worker's company limited Tianjin, Dihua recovery in Tianjin institute, 20080327 analytical pure); Glacial acetic acid (Beijing Chemical Plant, lot number 000108, analytical pure); The GSH-PX test kit all builds up bio-engineering research institute, lot number 20080416 available from Nanjing; Myeloperoxidase (MPO) (MPO), superoxide dismutase (SOD), oxidized form of glutathione peroxidase (GSH-PX), malonaldehyde (MDA) and Coomassie brilliant blue test kit all build up bio-engineering research institute, reagent lot number 20090730 available from Nanjing.
1.3 key instrument TGL-16 type low-temperature and high-speed centrifuge, Anting Scientific Instrument Factory, Shanghai makes; DY-2 type electric driven glass refiner, NingBo XinZhi Biology Science Co., Ltd makes; 722S type uv-spectrophotometric instrument, Shanghai Precision Scientific Apparatus Co., Ltd makes KA-1000 type desk centrifuge Anting Scientific Instrument Factory, Shanghai and makes.
2 methods
2.1 70 of grouping rats are divided into 6 groups at random by body weight, are respectively normal control group (8), model control group (14), positive controls (12) and ginsenoside Rd's low dose group (12), middle dosage group (12), high dose group (12).
2.2 acute ulcer colitis Preparation of model: each is organized the rat fasting and can't help water 48h, record body weight, lumbar injection (i.p) pentobarbital sodium 40mg/kg anesthesia.After the anesthesia, rat head down, anus is inverted up, slowly inserts rat colon portion (degree of depth for apart from anus 8cm place) with irritating stomach pin (No. 16), slowly injects TNBS/50% alcoholic solution (0.4ml/100g rat body weight, TNBS 100mg/kg rat body weight).Inject about 1min consuming time (preventing during injecting that rat abdomen is extruded).Proinflammatory agent is injected the back that finishes and is slowly extracted filling stomach pin out, clamps the rat anus with wood clamp, hangs the rat tail, make whole Mus body tilt about 60 °, keep about 30min, take off clip, rat put return in the cage, keeping the rat position be a low buttocks high state, treats that nature regains consciousness.Rats in normal control group is slowly injected isopyknic normal saline respectively, and all the other operations are the same.
After 2.3 administration causes scorching 24h, normal control group, model control group are irritated stomach (1ml/100g) with water for injection, the positive controls rat gives SASP (30mg/kg, the 1ml/100g rat body weight) solution is irritated stomach, by ginsenoside Rd 20,40,80mg/kg, 1ml/100g irritates stomach to the basic, normal, high dosage group of Rd rat respectively.Successive administration 7d, administration every day 1 time.
2.4 the evaluation of colitis, the collection of specimen and processing
2.4.1 survey body weight every day, observe diarrhoea and feces character, ergasia, appetite etc.
2.4.2 rat is behind administration 7d, lumbar injection pentobarbital sodium 30mg/kg anesthesia, abdominal aortic blood, take out anus to distal colon (ileocecal valve level, do not contain caecum and vermiform appendix) whole colon and rectum section, cut off enteric cavity along the mesentery edge, wash down intestinal contents with cold saline, be tiled in (intestinal mucosa faces up) on the plastic plate after wiping away totally with filter paper, claim colon weight in wet base, the amount intestinal segment length of getting after marking substantially by the listed standards of grading of table 1.Get a fritter diseased region colon, be tiled on the cardboard and then use 4% formaldehyde fixed, paraffin embedding, HE dyeing is pressed the listed standards of grading of table 2 (with reference to the standards of grading of propositions such as Dieleman, and making an amendment slightly) and is carried out histological score.Getting part diseased region colon more accurately weighs, shred with eye scissors, add cold saline, make 10% tissue homogenate under the low temperature, get the not centrifugal homogenate tissue of part in order to surveying MPO, remainder with the centrifugal 10min of 3000r/min, get supernatant,-20 ℃ of refrigerators are preserved every biochemical indicators such as MDA to be measured, SOD, GSH-PX.Behind 4 ℃ of placements of the blood that takes out 2h, with the centrifugal 10min of 3000r/min, separation of serum ,-20 ℃ of refrigerators are preserved biochemical indicator to be measured.
Table 1 colon cardinal principle and Histological injury's standards of grading
2.4.3 detect the mensuration of index
1) myeloperoxidase (MPO) (MPO) vitality test in the tissue: use spectrophotometry.This enzyme is the marrow peroxidase, and this enzyme has the ability that makes hydrogen-peroxide reduction, utilizes the vigor that these characteristics can enzyme analysis, and the number of quantitative assay neutrophilic leukocyte.Get not centrifugal 10% colon homogenate 90 μ l, operate, generate yellow compound after connecting the anisidine hydrogen supply by hydrogen donor is adjacent by the test kit description, at the 460nm place by the colorimetric determination absorbance, thereby extrapolate the MPO enzyme activity unit.Enzyme activity unit is defined as in the tissue: every gram tissue sheet H in 37 ℃ reaction system that wets
2O
2The 1 μ mol that is decomposed is an enzyme activity unit.
MPO vitality test in the serum: measuring principle is the same, gets serum 90 μ l, operate by the test kit description, at the 460nm place by the colorimetric determination absorbance, thereby extrapolate the MPO enzyme activity unit.Enzyme activity unit is defined as in the serum: every liter of serum H in 37 ℃ reaction system
2O
2The 1 μ mol that is decomposed is 1 enzyme activity unit.
2) Rd is to the influence of UC rat colon tissue oxidizing damage and oxidation resistance:
Malonaldehyde in the tissue (MDA) assay: MDA is a lipid peroxide, with thiobarbituricacid (TBA) colorimetric method for determining.Lipid peroxide catabolite malonaldehyde can with the thiobarbituricacid condensation, form red product, at the 532nm place maximum absorption band is arranged.Get 10% colon homogenate supernatant, 200 μ l and measure, represent with every milligram of histone nanomole number (nmol/mgprot).
Superoxide dismutase in the tissue (SOD) vitality test: detect with xanthine-xanthine oxidase.Produce ultra-oxygen anion free radical by xanthine-xanthine oxidase response system, latter's oxidation azanol forms nitrite, presents aubergine under the effect of developer, measures its absorbance at the 550nm place with visible spectrophotometer.When containing SOD in the sample, then superoxide anion there is the transitivity inhibitory action, the nitrite of formation is reduced, the absorbance of measuring pipe during colorimetric is lower than the absorbance of control tube, calculates the SOD vigor that can obtain in the sample by formula.Getting 1% colon's supernatant, 70 μ l operates by the test kit description.Total SOD unit of activity (U) is defined as in the tissue homogenate: it is a SOD unit of activity that every milligram of histone SOD suppression ratio in the 1ml reactant liquor reaches 50% o'clock pairing SOD amount.
Glutathion peroxidase in the tissue (GSH-PX) assay: GSH-Px can promote hydrogen peroxide (H
2O
2) generate H with reduced form GSH reaction
2O
2And oxidized form of glutathione (GSSG), the vigor of glutathion peroxidase can be represented with the speed of its enzymatic reaction, measures the consumption of reduced glutathion in this enzymatic reaction, then can obtain the vigor of enzyme.The vigor of GSH-Px is represented with the response speed of catalysis GSH, because these two substrates are not having under the condition of enzyme, also can carry out redox reaction (claiming non-enzymatic reaction), so must deduct the part that the caused GSH of non-enzymatic reaction reduces when calculating this enzyme activity at last.Get 10% colon homogenate supernatant, 60 μ l, press the operation of test kit description.
3) Rd influence that proinflammatory cytokine in the UC rat colon tissue is discharged:
TNF-α measures in the tissue: adopt the double-antibody sandwich elisa method to detect.
Operating procedure:
1. the dilution of standard substance: (360ng/L) is diluted to 240ng/L respectively with standard substance, 160ng/L, 80ng/L, 40ng/L, 5 concentration of 20ng/L.
2. application of sample: establish blank well (the blank hole does not add sample and enzyme marking reagent, and all the other each step operations are identical), testing sample hole respectively.Testing sample adds sample diluent 40 μ l in the air earlier on the enzyme mark wraps by plate, and then adds testing sample 10 μ l (the final diluted concentration of sample is 5 times).During application of sample sample is added on bottom, ELISA Plate hole, does not touch hole wall as far as possible, rock mixing gently.
3. incubation: with rearmounted 37 ℃ of incubations of shrouding film shrouding 30 minutes.
4. dosing: with standby after 30 times of dilutions of 30 times of concentrated cleaning solutions usefulness distilled water.
5. washing: carefully take the shrouding film off, discard liquid, dry, cleaning mixture is filled it up with in every hole, leaves standstill after 30 seconds to discard, and so repeats 5 times, pats dry.
6. enzyme-added: every hole adds enzyme marking reagent 50 μ l, except the blank well.
7. incubation: operation is with 3.
8. washing: operation is with 5.
9. colour developing: every hole adds developer A50 μ l earlier, adds developer B50 μ l again, the light shaking mixing, and 37 ℃ of lucifuges developed the color 15 minutes.
10. stop: every hole adds stop buffer 50 μ l, cessation reaction (this moment, blue upright commentaries on classics was yellow).
11. measure: with the blank well zeroing, the 450nm wavelength is measured the absorbance (OD value) in each hole in regular turn.Mensuration should be after adding stop buffer be carried out with interior in 15 minutes.
The mensuration of IL-1 β in the tissue: adopt the double-antibody sandwich elisa method to detect.
Operating procedure:
1. the dilution of standard substance: (45ng/L) is diluted to 30ng/L respectively with standard substance, 20ng/L, 10ng/L, 5ng/L, 5 concentration of 2.5ng/L.
2. application of sample: establish blank well (the blank hole does not add sample and enzyme marking reagent, and all the other each step operations are identical), testing sample hole respectively.Testing sample adds sample diluent 40 μ l in the air earlier on the enzyme mark wraps by plate, and then adds testing sample 10 μ l (the final diluted concentration of sample is 5 times).During application of sample sample is added on bottom, ELISA Plate hole, does not touch hole wall as far as possible, rock mixing gently.
3. incubation: with rearmounted 37 ℃ of incubations of shrouding film shrouding 30 minutes.
4. dosing: with standby after 30 times of dilutions of 30 times of concentrated cleaning solutions usefulness distilled water.
5. washing: carefully take the shrouding film off, discard liquid, dry, cleaning mixture is filled it up with in every hole, leaves standstill after 30 seconds to discard, and so repeats 5 times, pats dry.
6. enzyme-added: every hole adds enzyme marking reagent 50 μ l, except the blank well.
7. incubation: operation is with 3.
8. washing: operation is with 5.
9. colour developing: every hole adds developer A50 μ l earlier, adds developer B50 μ l again, the light shaking mixing, and 37 ℃ of lucifuges developed the color 15 minutes.
10. stop: every hole adds stop buffer 50 μ l, cessation reaction (this moment, blue upright commentaries on classics was yellow).
11. measure: with the blank well zeroing, the 450nm wavelength is measured the absorbance (OD value) in each hole in regular turn.Mensuration should be after adding stop buffer be carried out with interior in 15 minutes.
3) Rd is to pressing down the influence that inflammatory cytokines IL-10 discharges in the UC rat colon tissue:
Adopt the double-antibody sandwich elisa method to detect.
Operating procedure:
1. the dilution of standard substance: (72ng/L) is diluted to 48ng/L respectively with standard substance, 32ng/L, 16ng/L, 8ng/L, 5 concentration of 4ng/L.
2. application of sample: establish blank well (the blank hole does not add sample and enzyme marking reagent, and all the other each step operations are identical), testing sample hole respectively.Testing sample adds sample diluent 40 μ l in the air earlier on the enzyme mark wraps by plate, and then adds testing sample 10 μ l (the final diluted concentration of sample is 5 times).During application of sample sample is added on bottom, ELISA Plate hole, does not touch hole wall as far as possible, rock mixing gently.
3. incubation: with rearmounted 37 ℃ of incubations of shrouding film shrouding 30 minutes.
4. dosing: with standby after 30 times of dilutions of 30 times of concentrated cleaning solutions usefulness distilled water.
5. washing: carefully take the shrouding film off, discard liquid, dry, cleaning mixture is filled it up with in every hole, leaves standstill after 30 seconds to discard, and so repeats 5 times, pats dry.
6. enzyme-added: every hole adds enzyme marking reagent 50 μ l, except the blank well.
7. incubation: operation is with 3.
8. washing: operation is with 5.
9. colour developing: every hole adds developer A50 μ l earlier, adds developer B50 μ l again, the light shaking mixing, and 37 ℃ of lucifuges developed the color 15 minutes.
10. stop: every hole adds stop buffer 50 μ l, cessation reaction (this moment, blue upright commentaries on classics was yellow).
11. measure: with the blank well zeroing, the 450nm wavelength is measured the absorbance (OD value) in each hole in regular turn.Mensuration should be after adding stop buffer be carried out with interior in 15 minutes.
2.5 statistical procedures
Parametric test Student ' s t check, all (expression P＜0.01 of x ± s), P＜0.05 is for difference has statistical significance, and non parametric tests is with Mann-Whitney U check with mean ± standard deviation.
3. result
3.1 rat outward appearance sign overview is set up the rat ulcer colitis with TNBS-50% ethanol, animal in modeling 2d loose stool occurs, become thin, loss of appetite, roll up, hair not gloss, movable reduce etc.Each administration treated animal outward appearance symptom all is lighter than model control group.
3.2 the influence that the ginsenoside Rd changes rat body weight is at experimental session, normal control group body weight is increase trend, on average increases by 18.4%; The model control group body weight subtracts afterwards earlier and increases, but increasess slowly, and on average increases by 9.4%; SASP group, the basic, normal, high dosage group of GSPE body weight subtract afterwards earlier and increase, but it is fast than model control group to gather way, average respectively increase by 14.7%, 12.2%, 13.8% and 12.3%.Each group rat is put to death precursor recast statistical analysis, and the result shows that the model control group rat body weight is starkly lower than normal control group and each administration group, and difference has statistical significance (P＜0.01 or P＜0.05), the results are shown in Table 2
The influence that table 2 ginsenoside Rd changes the UC rat body weight (x ± s, n=8-14)
Model control group and normal control group compare:
*P＜0.01; Administration group and model control group compare:
#P＜0.05,
##P＜0.01
3.3 the ginsenoside Rd compares with the normal control group the influence that the rat colon weight in wet base changes, each administration group rat colon weight in wet base index all increases to some extent, model control group colon weight in wet base index (colon weight/colon length wherein, mg/mm) increase maximum, statistical significance (P＜0.01) is arranged with normal control group comparing difference; Each administration group and model control group relatively colon weight in wet base index reduce, and difference has statistical significance (P＜0.01 or P＜0.05), sees Table 3.
Table 3 ginsenoside Rd to the influence of UC rat colon weight in wet base index variation (x ± s, n=8-14)
Model control group and normal control group compare:
*P＜0.01; Each administration group and model control group compare:
#P＜0.05,
##P＜0.01
3.4 gross examination of skeletal muscle appraisal result
Compare with the normal control group, the damage of model control group colon obviously increases the weight of, and difference has statistical significance (P＜0.01), compares with model control group, and each administration group colon damage obviously alleviates, and difference has statistical significance (P＜0.01).Result's demonstration sees Table 4.
Table 4 ginsenoside Rd treats back UC rat colon anatomical lesion appraisal result (n=8-12)
Model control group and normal control group compare:
*P＜0.01; Each administration group and model control group compare:
##P＜0.01
3.5 histopathology appraisal result
Light microscopic is observed down, visible a large amount of inflammatory exudates and tissue necrosis in the model control group mucous membrane of colon tissue; Visible a large amount of re-epithelialize tissues of each dosage group ulcer surface of ginsenoside Rd and newborn body of gland; Compare with the normal control group, the damage of model control group colon is obvious, and difference has statistical significance (P＜0.01), compares with model control group, and each administration group rat colon degree of tissue damage alleviates, and difference has statistical significance (P＜0.01).The results are shown in Table 5.
Table 5 ginsenoside Rd treats back UC rat colon Histological injury's appraisal result (n=8-12)
Model control group and normal control group compare:
*P＜0.01; Each administration group and model control group compare:
##P＜0.01
3.6Rd influence to the leukocyte infiltration degree:
The MPO content value sees Table 6 in the rat blood serum 3.6.1Rd the active influence of MPO in rat blood serum experiment respectively organized, and by table as seen: with the comparison of normal control group, model control group MPO vigor obviously increases, and difference has statistical significance (P＜0.01); SASP administration group, MPO and model control group comparison vigor obviously reduce in basic, normal, high each dosage of Rd, and difference has statistical significance (P＜0.01 or P＜0.05).
The MPO content value sees Table 6 in the rat colon tissue 3.6.2Rd the active influence of MPO in rat colon tissue experiment respectively organized, and by table as seen: with the comparison of normal control group, model control group MPO vigor obviously increases, and difference has statistical significance (P＜0.01); SASP administration group, MPO vigor and the apparent in view reduction of model control group in basic, normal, high each dosage of Rd, difference has statistical significance (P＜0.01 or P＜0.05).
Table 6 ginsenoside Rd to the influence of MPO vigor in TNBS inductive UC rat colon tissue and the serum (x ± s, n=8-12)
Model control group and normal control group compare:
*P＜0.01,
*P＜0.05; Each administration group and model control group compare:
##P＜0.01,
#P＜0.05
3.7Rd influence to UC rat colon tissue oxidizing damage and oxidation resistance:
3.7.1Rd the influence to MDA content in the rat colon tissue: experiment organizes respectively that MDA assay value sees Table 7 in the rat colon tissue, by table as seen: compare with the normal control group, model control group MDA content obviously increases, and difference has statistical significance (P＜0.01); Compare with model control group, MDA content obviously reduces in each administration group colon, and difference has statistical significance (P＜0.01).
3.7.2Rd to the active influence of rat colon tissue GSH-PX: experiment organizes respectively that GSH-PX determination of activity value sees Table 7 in the rat colon tissue, by table as seen: compare with the normal control group, model control group GSH-PX is active obviously to be reduced, and difference has statistical significance (P＜0.01); SASP administration group, the GSH-PX activity obviously increases than model control group in the basic, normal, high dosage of Rd, and difference has statistical significance (P＜0.01).
3.7.3Rd to the active influence of rat colon tissue SOD: experiment organizes respectively that SOD determination of activity value sees Table 7 in the rat colon tissue, by table as seen: with the normal control group relatively, the active obvious reduction difference of model control group SOD has statistical significance (P＜0.05); SASP administration group, the GSH-PX activity obviously increases than model control group in the basic, normal, high dosage of Rd, and difference has statistical significance (P＜0.01).
Table 7 ginsenoside Rd to the influence of the inductive rat UC of the TNBS MDA of colon content, GSH-Px and SOD vigor (x ± s, n=8-12)
3.8Rd influence to proinflammatory cytokine expression in the UC rat colon tissue
TNF-alpha expression level determination value sees Table 8 in the rat colon tissue 3.8.1Rd the influence of rat colon tissue T NF-alpha expression level experiment respectively organized, by table as seen: compare with the normal control group, model control group TNF-alpha expression level significantly increases (P＜0.01); SASP administration group, TNF-alpha expression level obviously reduces (P＜0.01) than model control group in Rd low dosage, the middle and high dosage.
3.8.2Rd rat colon is organized the influence of IL-1 β expression: experiment organizes respectively that IL-1 β level determination value sees Table 8 in the rat colon tissue, by table as seen: compare with the normal control group, model control group IL-1 β expression obviously increases, and difference has statistical significance (P＜0.01); SASP administration group, IL-1 β expression obviously reduces than model control group in the basic, normal, high dosage of Rd, and difference has statistical significance (P＜0.01).3.9Rd to pressing down the influence of inflammatory cytokines IL-10 level in the UC rat colon tissue: experiment organizes respectively that IL-10 level determination value sees Table 8 in the rat colon tissue, by table as seen: compare with the normal control group, model control group IL-10 expression significantly reduces (P＜0.01); SASP administration group, the IL-10 expression obviously increases (P＜0.05) than model control group in Rd low dose group and the high dose.Among the Rd dosage group IL-10 expression value compare with model control group increase trend but difference does not have statistical significance.
Table 8 ginsenoside Rd to the influence of the inductive rat UC of the TNBS IL-1 β of colon, TNF-α, IL-10 expression (x ± s, n=6-12)
Model control group and normal control group compare:
*P＜0.01,
*P＜0.05; Each administration group and model control group compare:
##P＜0.01,
#P＜0.05
4. conclusion
Recently Chinese scholars has been carried out big quantity research for the ginsenoside, discover that these saponins active component have blood system, cardiovascular system, nervous system, immune system, substance metabolism system, and various pharmacological activities such as antiinflammatory, defying age, antitumor.This subject study to as if one of its main active ginsenoside Rd, studied its therapeutical effect, and its antiinflammatory mechanism carried out preliminary study for the alcohol induced rat acute ulcerative colitis of TNBS-50%.Rat colon weight in wet base index (colon weight/length) is considered to indicate the comparatively stable of the inductive experimental rat ulcerative colitis of the TNBS-50% order of severity and one of index reliably, and this result of study shows the significantly reduction of comparing with model control group of various dose ginsenoside Rd administration group rat colon weight in wet base index.The ginsenoside Rd can also prevent that the inductive UC rat body weight of TNBS-50% from descending; From substantially and histological scores can see that also the ginsenoside Rd can alleviate general form and learn and the pathology pathological changes, significantly reduced tissue necrosis and ulcer formation, alleviate inflammatory cell infiltration, the reparation of promotion epithelial cell and ulcer surface.
The generation of the result of study metabolite that confirmed inflammatory mediator and active oxygen and inflammatory bowel both at home and abroad and develop relevant.A large amount of zooperies confirms that also the metabolite of active oxygen may be inseparable with the generation of inflammation, and result of study shows that the metabolite of active oxygens excessive, a large amount of in the inflammatory enteritis patient mucosa may be relevant with the morbidity of this type of disease simultaneously.These metabolites may be one of inducements that causes the patients of ulcerative colitis mucosa injury.Phagocyte is its main source, and a large amount of infiltration intestinal mucosa of these cells between the inflammation period of disease stress takes place discharge a large amount of active oxygens, and causes the oxidative damage of intestinal mucosa.Although intestinal mucosa has the defense mechanism of good anti-oxidative damage, to compare ability lower with other organ of body such as liver and lung.The excessive release of oxidative stress product may destroy the mucosal defense mechanisms of intestinal tract itself.At present, the increase of excessive active oxygen that stress produce owing to phagocyte in the intestinal mucosa and lipid peroxide is as the index of describing inflammatory enteritis patient's biopsy specimen (IBD).Indivedual researcheres find that also the ginsenoside Rd has the immunological adjuvant activity, and can excite and balance Th1 and Th2 immune response by regulating production of cytokines and expressing.Because cytokine has critical role in the immune system regulation and control, the immunology pathogenesis of present UC particularly cytokine pathogenesis more and more is subject to people's attention.And pro-inflammatory cytokine and press down the important pathogenesis that dysimmunity due to the dysequilibrium between the inflammatory cytokine is regarded as UC.The present invention has confirmed that also the ginsenoside Rd is removing oxygen-derived free radicals, antioxidation, the effect of immunity regulatin remedy, and we can further inquire into and sum up from the following aspects for the mechanism of action of rat ulcer colitis for it.
One, Rd is to the influence of leukocyte infiltration degree
1. marrow peroxidase (MPO): be the higher a kind of enzyme of content in the neutrophilic granulocyte, increasing of its content can reflect that neutrophilic granulocyte soaks in a certain tissue, the existence of reflection inflammation in tissue indirectly, and the MPO level can reflect that the infiltration number and the activity of neutrophilic granulocyte, MPO have been used as one of index of estimating the enteritis order of severity in the inflammation group
[21,22]In this experiment in rats in normal control group colon and the serum sample MPO vigor minimum in each group, model control group MPO activity is the highest in each group, this has illustrated that also the damage of model control group neutrophil infiltration and enteritis is the most serious.Compare with model control group, the SASP group, the MPO in the basic, normal, high dosage group of the ginsenoside Rd UC rat colon tissue is active obviously to be reduced (being dose-dependence), and [18] MPO testing result conforms to pathology tissue detection result.Illustrate that Rd can alleviate neutrophil infiltration degree, amelioration of inflammation.
Two, Rd is to the influence of UC rat colon tissue oxidizing damage and oxidation resistance
1. malonaldehyde (MDA) and superoxide dismutase (SOD): body is by enzyme system and non-enzyme system generation oxygen-derived free radicals, the latter can attack the polyunsaturated fatty acid (PUFA) in the biomembrane, cause lipid peroxidation, therefore and form lipid peroxide, as malonaldehyde (MDA), hydroxyl, carbonyl, ketone group, hydroperoxy or interior peroxy, and new oxygen-derived free radicals etc.Oxygen-derived free radicals not only causes cell injury by the peroxidating of polyunsaturated fatty acid in the biomembrane, and can also cause cell injury by the catabolite of fat hydroperoxides.Thereby the amount of test MDA can reflect the snperoxiaized degree of body inner lipid, the indirect cells injury degree that reflects; And MDA and SOD have close ties, the height indirect reaction of SOD vigor body remove the ability of oxygen-derived free radicals, and the MDA body cell order of severity that attacked by free radical that got the height indirect reaction.In this experiment, the model control group rat colon organizes the normal matched group of MDA content obviously to raise, and the SOD vigor obviously reduces, and the MDA content that each medication group is compared with model control group in the colon obviously reduces, and the SOD vigor significantly raises and is dose-dependence.This result shows that Rd has stronger antioxidation in treatment ulcerative colitis damage, and its antiinflammatory action may be with to alleviate lipid peroxidation relevant.
2. glutathion peroxidase (GSH-Px): be a kind of important peroxide breakdown enzyme that extensively exists in the body, its physiological function mainly is that catalysis GSH participates in peroxidization, peroxide that removing produces in the Cellular respiration metabolic process and hydroxy radical, thus the peroxidation of cell membrane polyunsaturated fatty acid alleviated.Thereby the structure of protection cell membrane and interference and the infringement that function is not subjected to peroxide.Therefore the active variation of GSH-Px also can reflect the variation of antioxidant ability of organism.In this experiment, the normal matched group of model control group rat colon tissue GSH-PX vigor significantly reduces, the GSH-PX vigor obviously raises and SASP organizes and the basic, normal, high dosage of ginsenoside Rd is compared with model control group, difference has statistical significance, and this result has further confirmed the oxidation resistance that the ginsenoside Rd is stronger.
Three, the equilibrated influence of ginsenoside Rd's pair cell factor
1. suppress the expression of proinflammatory factor
IL-1 β is a kind of proinflammatory factor, and it and cytokines such as TNF-α, IL-6, IL-8 play an important role in inducing the mucous membrane of colon inflammation, and participates in continuing and amplification of inflammation.It increases the weight of inflammatory reaction by the immune cell activated cascade reaction.Diarrhoea is the cardinal symptom of enteritis, and the IL-1 β main stimulus object of diarrheal seemingly.High dose IL-1 β causes epithelial cell necrosis, edema, neutrophil infiltration and goblet cell disappearance.Think that at present its morbidity effect in UC has aspect 4: activatory macrophage can discharge IL-1 β in (1) UC proper mucous membrane, activates dendritic cell, engulfs, digests exotic antigen, and the released antigen fragment is also presented to T lymphocyte generation immunoreation; (2) IL-1 β can increase cytokine such as IL-6, TNF-α and the IL-8 by macrophage produced, make neutrophilic granulocyte assemble to inflammation part, enter the intestinal diseased region, thereby cause a series of intestinal pathological changes, as the damage of colonic epithelium, polyangitis, crypt abscess etc., finally cause the morbidity of UC; (3) the unbalance of IL-1 β and IL-1 receptor is the important step that UC takes place; (4) IL-1 β can discharge H by inducing
2O
2, the Ca of influence
2+The switch of the signal transduction pathway of release and NK-A, thus UC patient's colonic smooth muscle contractile function disorder caused.In this experiment in the model control group rat colon tissue the normal matched group of IL-1 β expression significantly raise, and each dosage group of ginsenoside Rd can significantly reduce IL-1 β expression in the ulcerative colitis rat colon tissue, illustrates that it is to reduce the IL-1 β expression of diseased region that the ginsenoside Rd alleviates one of main mechanism of UC tissue injury.
Tumor necrosis factor (TNF-α) is a kind of important short inflammatory factor and immunoregulatory factor, a kind of cytokine that participates in the UC morbidity by activatory macrophage and T lymphocytic emiocytosis, TNF-α causes that the mechanism of intestinal mucosa damage comprises the release platelet activating factor, generates leukotriene and oxygen-derived free radicals, interior inducible nitric oxide synthase (iNOS) mRNA of inducing cell expresses and the generation of the interior tetrahydrobiopterin of cell (the short factor of giving birth to of a kind of NO), makes the NO generation increase and cause cells injury; The hypertrophy of TNF-α scalable neutrophilic granulocyte and macrophage, maturation and activation, and promote its adhesion, migration and take off granule, strengthen their killing ability of engulfing, promote them to discharge inflammatory protein and inflammatory mediator, participate in the damage of intestinal mucosa directly; Vascular endothelial cell adhesion molecule that TNF-α raises and the expression of chemotactic cytokine IL-8 and increase the gathering of inflammatory cell; TNF-α also can induce the generation of IL-1 β and cause the damage of intestinal mucosa; TNF-α too much also can cause intestinal endolymph apoptosis by activating Caspase (Caspases), reduces the immunologic function of intestinal mucosa.TNF-α is bringing into play important effect in the inductive colitis pathogenesis of TNBS, TNF-α is one and has remarkable chemotactic activity cytokine, under the synergism of IL-1 β, relevant enzyme is activated (comprise nitricoxide synthase, phospholipase A, Cycloxygenase and protease etc.), thereby induce adhesion molecule and other inflammatory cytokines to express, directly or indirectly cause mucosa injury, these cytokines are the reaction important biological albumen of regulating and control inflammation, and are closely related with the pathogenesis of ulcerative colitis (UC).The normal matched group of model control group rat colon tissue T NF-alpha content significantly raises in this experiment, and each administration group of ginsenoside Rd can significantly reduce TNF-alpha expression level in the ulcerative colitis rat colon tissue.This result shows that the ginsenoside Rd treats the inductive rat acute ulcerative colitis of TNBS and has notable therapeutic effect and its can reduce in the tissue pro-inflammatory cytokine TNF-alpha expression and to regulate immunologic balance closely related.
2. to pressing down the regulating and controlling effect of the scorching factor
Interleukin-11 0 (IL-10) is that discoveries such as Fiorentino in 1989 have the inflammation SC factor of multi-efficiency, the reaction of participation panimmunity, its main biologic activity is an immunosuppressive action, short inflammatory factor of generations such as mononuclear cell, neutrophilic granulocyte, eosinophilic granulocyte and chemotactic factor be can suppress and IL-1 β, L-6, L-8, TNF-α, the generation of granulocyte-macrophage colony stimutaing factor (GM-CSF) etc. suppressed; The propagation that stops antigen-specific T cell, suppress the expression of histocompatibility complex (MHC)-II quasi-molecule and some stimulating factor such as the B7 on antigen presenting cell (APC) surface, reduce the antigen presentation ability of APC, the expression of downward modulation effector lymphocyte's IL-2mRNA, suppress the generation of IL-2, suppress the synthetic of inflammatory mediator nitric oxide, oxygen-derived free radicals, prostaglandin etc.; Suppress the inflammation relevant enzyme iNOS (inducible nitric oxide synthase) of macrophage and the expression of COX-2 (cyclooxygenase); By suppressing the generation suppression of natural killer NK cell activity of IFN-γ, suppress the Th0 cell to the Th1 transformation, the migration of inflammation-inhibiting cell.Can also suppress expression by the inductive tissue factor of LPS, thus anticoagulant system activity.IL-10 also has immunostimulation in addition, can promote the propagation and the differentiation of B cell, produce a large amount of IgG, IgA, IgM, also can increase the survival rate of static B cell, in the startup of tissue specificity autoimmunity damage and development, play key regulating action.
1993, Kuhn, Lohler, Rennick, the mice of researcheres such as Rajewsky and Muller discovery IL-10 gene knockout can be brought out enteritis, and the variation of IL-10 is consistent with ulcerative colitis change of illness state degree, and is also consistent with therapeutic effect.The model control group rat colon organizes the normal matched group of IL-10 content significantly to reduce in this experiment, the IL-10 content that ginsenoside Rd's low dose group, high dose are compared with model control group in the colon significantly increases, and this result shows that Rd can promote to press down the expression of scorching factor IL-10 significantly.
By The above results as can be known, the ginsenoside Rd treats the inductive rat acute ulcerative colitis of TNBS/50% has one of its dominant mechanism of obvious curative effects to be: can suppress the generation of proinflammatory cytokine IL-1 β, TNF-α significantly, and the expression that promotes anti-inflammatory cytokines IL-10.
Four, with the relation of calcium channel
The current research result confirms that the ginsenoside Rd is a novel non-potential dependent calcium ion channel blocker, and current potential dependent/non-dependent calcium channel is by irritation cell membrane receptor (membrane receptor calcium activate channel, ROCC) or the sarcoplasmic reticulum calcium ion storage pool decay (calcium channel that the calcium pond is handled, SOC) finish the adjusting function of intracellular calcium, and for various kinds of cell epithelial cell for example, the lymphocytic function of T plays very important regulating and controlling effect.Inflammatory cell belongs to non-excitatory cells, and SOC is the main thoroughfare of stream in the outer calcium of these cells born of the same parents.The rising of calcium concentration can make inflammatory cell in the Cytoplasm, comprises neutrophil activation or plays a role, and can make relevant enzyme and inflammatory mediator activate, discharge.Regulate Ca in the neutrophilic granulocyte kytoplasm
2+The intensity of concentration scalable inflammatory reaction.Calcium antagonist has stable mast cell membrane and stops the generation of taking off granule, suppressing lymphocyte transformation and IL-2, reduces the effect of epidermal langerhan cell number, and calcium channel blocker such as nifedipine, verapamil can be blocked calcium channel, reduce Ca in the born of the same parents
2+Concentration suppresses mast cell degranulation, suppresses releasing and activity of inflammatory cytokines and release.But this type of medicine such as nifedipine block voltage dependent channel (PDC), and do not influence SOC.Because inflammatory cell belongs to non-excitatory cells, the SOC inhibitor is more special than the PDC blocade to the calcium channel on the inflammatory cell, should be able to more effectively hinder the rising of calcium concentration in the inflammatory cell and inflammation is produced inhibitory action.The SOC blocker of having found has cation, cytochrome P-450 inhibitor, arachidonic acid metabolic enzyme inhibitor etc., but they lack ideal specificity mostly, or only is the instrument medicine.As previously mentioned, existing anti-inflammatory agent such as aspirin class, glucocorticoids and and anti-UC medicine sulfasalazine all do not have a calcium antagonism yet.Regulate inflammatory reaction with specific SOC blocker a new target spot will be provided for the control of inflammation, the research that searching can be treated the specific SOC blocker of UC inflammation has important theory to be worth and using value.
Embodiment 2: the ginsenoside Rd treats the best route of administration of rat acute ulcerative colitis
Method: set up rat acute ulcerative colitis model with inflammation-induced agent trinitro-benzene-sulfonic acid (TNBS)/50% alcoholic solution revulsion.Rat is divided into model control group, Rd gastric infusion group, Rd intramuscular injection group at random.The Rd dosage is 30mg/kg, administration every day 1 time, successive administration 7 days.The concrete detection method of every index is seen embodiment 1.
The result: 1. with the model control group ratio, Rd gastric infusion group and Rd intramuscular injection group rat colon weight in wet base index all obviously alleviate, and difference has statistical significance (P＜0.01 or P＜0.05), and wherein gastric infusion group rat colon weight in wet base index is minimum.
2. compare with model control group, Rd gastric infusion and Rd intramuscular injection can obviously reduce scoring of UC rat colon gross examination of skeletal muscle and histopathology scoring, and pathology damage (P＜0.01) difference that alleviates rat colon has statistical significance.
Conclusion: the ginsenoside Rd has the obvious treatment effect to the inductive rat ulcer colitis of TNBS, and the gastric infusion mode is the route of administration of treatment ulcerative colitis the best.
Claims (7)
1. the ginsenoside Rd is in the application of preparation treatment colitis medicine.
2. ginsenoside Rd as claimed in claim 1 is characterized in that the application of ginsenoside Rd at preparation treatment ulcerative colitis medicine in the application of preparation treatment colitis medicine.
3. ginsenoside Rd as claimed in claim 1 is characterized in that the application of ginsenoside Rd at preparation treatment non-bacterial chronic colitis medicine in the application of preparation treatment colitis medicine.
4. preparing the application for the treatment of the colitis medicine as claim 1 or 2 or 3 described ginsenoside Rds, it is characterized in that of the application of ginsenoside Rd's oral formulations at preparation treatment ulcerative colitis medicine.
5. ginsenoside Rd as claimed in claim 4 is characterized in that in the application of preparation treatment colitis medicine described oral formulations is capsule, microcapsule, liposome, granule, tablet, oral liquid.
6. as claim 1-3,5 one of any described ginsenoside Rds application at preparation treatment colitis medicine, it is characterized in that the ginsenoside Rd is the ginsenoside Rd's monomeric compound that extracts from Radix Notoginseng, wherein ginsenoside Rd's monomer is more than 95%; Also comprise ginsenoside Rd's monomeric compound that extraction separation obtains root from araliaceae ginseng plant's Radix Ginseng, Radix Panacis Quinquefolii, stem, leaf, flower, the fruit.
7. ginsenoside Rd as claimed in claim 4 is characterized in that ginsenoside Rd's monomeric compound that the ginsenoside Rd extracts in the application of preparation treatment colitis medicine from Radix Notoginseng, wherein ginsenoside Rd's monomer is more than 95%; Also comprise ginsenoside Rd's monomeric compound that extraction separation obtains root from araliaceae ginseng plant's Radix Ginseng, Radix Panacis Quinquefolii, stem, leaf, flower, the fruit.
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| WO2015021922A1 (en) * | 2013-08-13 | 2015-02-19 | 广东泰禾医药科技有限公司 | Ginsenoside rd or enteric formulation thereof and use |
| CN104706646A (en) * | 2015-02-10 | 2015-06-17 | 甘肃省人民医院 | Application of ginsenoside Rg3 in preparation of drugs for ulcerative colitis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2015021922A1 (en) * | 2013-08-13 | 2015-02-19 | 广东泰禾医药科技有限公司 | Ginsenoside rd or enteric formulation thereof and use |
| CN104367585A (en) * | 2013-08-13 | 2015-02-25 | 广东泰禾医药科技有限公司 | Ginsenoside Rd/ginsenoside Rd enteric preparation and applications thereof |
| CN104706646A (en) * | 2015-02-10 | 2015-06-17 | 甘肃省人民医院 | Application of ginsenoside Rg3 in preparation of drugs for ulcerative colitis |
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