CN105535011A - Applications of mushroom mycelium polysaccharides - Google Patents
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Abstract
The present invention provides applications of mushroom mycelium polysaccharides in preparation of drugs for treatment of inflammatory bowel diseases. According to the present invention, calculation is performed by adopting the colorectal mucosa tissue score of less than 5 as the cure, wherein the cure rate of the mushroom mycelium polysaccharides on the 5% DSS-induced chronic ulcerative colitis in the acute phase is 43.8%, the cure rate of the mushroom mycelium polysaccharides on the 1% DSS-induced chronic ulcerative colitis in the chronic phase is 60%, and the cure rate of the mushroom mycelium polysaccharides on the TNBS-induced Crohn's disease is 72.8%; and the provided mushroom mycelium polysaccharides can effectively treat ulcerative colitis and Crohn's disease.
Description
Technical field
The present invention relates to medicinal chemistry art, particularly, relate to the application of Lentinula edodes mycelium polysaccharide.
Background technology
Inflammatory bowel (inflammatoryboweldisease, IBD) be the colorectal mucosa chronic nonspecific inflammation that the cause of disease and pathogenesis are still not clear, with stomachache, diarrhoea, mucopurulent bloody stool and tenesmus etc. for main clinical manifestation, it is the inflammatory bowel disease that is pathological characteristic with mucous membrane of colon chronic inflammatory disease and ulcer.In recent years because social pressure increases, dietary structure and living habit change, and its sickness rate is in increasing trend rapidly.Inflammatory bowel comprises ulcerative colitis (UlcerativeColitis, UC) and Crohn disease (Crohn ' sDisease, CD).The mucosa diffusivity hyperemia that the pathological characteristic of ulcerative colitis (UlcerativeColitis, UC) is rectum, sigmoid colon position is master, edema, erosion and fester are formed, and pathological changes also may extend to proximal colonic.The pathological changes of UC mainly involves colon mucosa and tela submucosa, and scope is many from section colon far away, can drive in the wrong direction to nearly section and develop, even involve total colectomy and terminal ileum, distribute in seriality.Clinically in the chronic course of disease, its relapse rate is up to 71%, the ratio that simultaneously colon cancer occurs patient obviously increases, prognosis is poor, and there is no specificity at present and the little medicine of side effect, though clinical conventional medicine sulfasalazine, glucocorticoid, immunosuppressant etc. obtain certain curative effect, drug side effect greatly and still some patient is dissatisfied to curative effect, is refractory disease common in digestive system disease.
The pathogenesis of inflammatory bowel it be not immediately clear; but evidence show that the generation of IBD is relevant with immunity, heredity, environment and infective agent; immune factor plays an important role wherein; pro-inflammatory cytokine TNF-O, IL-6, IL-8 etc. as immunomodulator secreted by the lymphocyte activated, monokaryon one macrophage take part in the initial sum evolution of intestinal mucosa inflammatory reaction, play an important role in the reaction of normal or abnormal intestinal immune.Think that genetic predisposition, mucosal immunity and intestine microenvironment may take part in generation and the progress of its disease jointly at present.
The current treatment to this colitis is mainly based on coloclysis.Coloclysis can make medicine directly contact with diseased region and reduce the systemic side effects of medicine.Medicine at present for coloclysis mainly contains sulfasalazine, various glucocorticoid and some immunosuppressant.The concentration (20 ~ 40 μ g/ml) of its serum sulfapyridine of sulfasalazine and metabolite thereof is relevant with toxicity, and concentration is more than tool toxicity during 50 μ g/ml; Though glucocorticoid and immunosuppressant have certain curative effect drug side effect greatly and still some patient's curative effect is dissatisfied, some drugs can be absorbed into blood by the mucosa of inflammation and patient's edema, blood glucose therefore can be caused to raise.Therefore highly effective and safe is studied and the suitable medicine of price has very important significance to colitis disease.In recent years, also have many research worker to advocate treatment by Chinese herbs colitis, but treatment by Chinese herbs method is partial to disperse, its classification diagnosis, dialectically treat the criterion lacking objective, microcosmic and qualitative, quantitative, zoopery is rarely seen to conventional index and pathological observation, still owes deeply; And the composition of compound preparation is complicated, be difficult to the clear and definite main component that it plays a role.Therefore in the urgent need to the medicine of the little and cheap treatment inflammatory bowel of exploitation determined curative effect, side effect.
Polysaccharide is that a class comes from higher plant, animal cell membrane and microorganism wall, the polysaccharide be polymerized by more than 10 monosaccharide, it is the natural polymer be formed by connecting by glycosides chain by aldose or ketose, its molecular weight be generally tens thousand of even reach millions of, polysaccharide is the polysaccharose substance that a class has bio-physiological activity and special health care, is one of four large base substances forming life.It not only provides framing structure and energy source for biology, also the adjustment of the various physiological process of wide participation cell.Within 1958, first Japanese scholars isolates the antitumor polysaccharide with beta glucan structure from Lentinus Edodes, after this, the research work of polysaccharide is in full swing, and people constantly obtain various polysaccharide from animal, plant, microorganism, and furthers investigate its biological activity.Research finds, the topmost biological activity of polysaccharide is embodied in the adjustment to body immune system function.Polysaccharide comprises Organism immunoregulation mechanism of action: activating macrophage and natural killer cell make it produce relevant cytokine; Activation of B lymphocytes and T lymphocyte; Activating complement system.
The effect only have the righting in differential diagnosis in tcm theory through the lentinan of water, alcohol extraction from mushroom fruiting body, consolidating, main component is glucosan peptide, and molecular weight is about 40-80 ten thousand.Lentinula edodes mycelium polysaccharide is adopt the mycelium of special mushroom strain to extract the obtained a kind of important polysaccharide biological response modifier with physiologically active after deep-layer liquid is cultivated.The main component of Lentinula edodes mycelium polysaccharide is mannatide, i.e. the mannose of α bonding and the coalition of peptide.Its saccharide portion, based on mannose, is secondly galactose, glucose, xylose and fucose; Peptide chain is then made up of 18 kinds of L-type aminoacid such as aspartic acid, serine, lysine, glutamic acid.Be mainly used in the various diseases caused because autoimmune function is low at present clinically, and for chronic viral hepatitis, liver protecting therapy; Also can as the ancillary drug of chemotherapy of tumors, determined curative effect.But, there is no relevant report about the application of Lentinula edodes mycelium polysaccharide in inflammatory bowel at present.Therefore extremely urgent at the medicine of the treatment inflammatory bowel of understanding development of new on its pathogenetic basis in depth.
Summary of the invention
For the defect that the curative effect of medication being used for the treatment of inflammatory bowel in prior art is not good, the invention provides the application of a kind of Lentinula edodes mycelium polysaccharide in the medicine of preparation treatment inflammatory bowel.
In above-mentioned application, the application of Lentinula edodes mycelium polysaccharide in the medicine of preparation treatment inflammatory bowel comprises: the application of Lentinula edodes mycelium polysaccharide in the medicine preparing treatment ulcerative colitis and Crohn disease.
In above-mentioned application, the polyoses content in Lentinula edodes mycelium polysaccharide is more than 60 % by weight.
In above-mentioned application, Lentinula edodes mycelium polysaccharide is 0.83 ~ 3.32mg/kg/d to the dose therapeutically effective of people.
In above-mentioned application, the pharmaceutical pack for the treatment of inflammatory bowel is containing Lentinula edodes mycelium polysaccharide, Lentinula edodes mycelium polysaccharide pharmaceutically acceptable salt, ester, hydrate or their combination and adjuvant.
In above-mentioned application, the medicine for the treatment of inflammatory bowel is selected from tablet, capsule, pill, suppository, aerosol, oral liquid, granule, powder, injection, syrup, medicated wine, tincture, distillate medicinal water, membrane or their combination.
In above-mentioned application, the administering mode of medicine for the treatment of inflammatory bowel comprises oral, injection, implantation, external, spraying, suction or their combination.
Different from the lentinan through water, alcohol extraction from mushroom fruiting body, Lentinula edodes mycelium polysaccharide provided by the invention has significant biological activity, its the similar a kind of host immune reinforcing agent of effect, the maturation of key cells, Differentiation and proliferation can be stimulated, improve host body balance, reach and recover or improve host cell to the reactivity effect of lymphokine, hormone and other biological active factors.The present inventor find the adjustment that the activation of NF-κ B is transcribed inflammation-related gene such as IL-8, IL6 and inflammatory bowel in close relations.This result of study shows, Lentinula edodes mycelium polysaccharide can suppress the nuclear translocation of NF-κ B in inflammatory bowel disease models mice intestinal submucosa tissue, and can reduce the content of IL-8 and IL-12 in serum, thus effectively can treat inflammatory bowel.
Lentinula edodes mycelium polysaccharide provided by the invention adopts special mushroom strain to ferment, be different from the righting of lentinan, consolidate effect, there is good biological activity, by regulating the immunologic function of body, make the immune state of intestinal local recover normal, and suppress inflammatory cell to discharge inflammatory factor, reduce the content of IL-8 and IL-12 in serum, suppress the inflammatory reaction of intestinal local disorders, final generation and progress of intervening the inflammatory bowel course of disease; Obviously can alleviate inflammation, the edema degree of chronic ulcerative colitis chronic phase, acute stage and Crohn disease mice Colon and rectum mucosa, and effectively can repair the human colorectal mucosa injury of diseased mice, make ulcer healing, obviously reduce the damage range of diseased mice Colon and rectum mucosa, and effectively can treat the diarrhoea of diseased mice.Therefore, Lentinula edodes mycelium polysaccharide provided by the invention can treat ulcerative colitis and Crohn disease effectively.
The advantage of inventing the treatment inflammatory bowel medicine provided is: this mechanism of drug action is clear and definite, drug effect is definite, and have good application prospect, patient is not only acceptant, and cheap, wide material sources, easily to obtain, be also convenient to understand patient to the reaction of medicine.In addition, lentinan will change the market structure of existing chronic enteritis medicine, become a kind of can the low clinical medicine of long-term taking, safety, side effect.
Accompanying drawing explanation
Fig. 1 is the schematic diagram of Lentinula edodes mycelium polysaccharide on the impact of colitis acute stage Colon and rectum length, cardinal principle and lesion tissue that DSS induces;
Figure 1A is the schematic diagram of Lentinula edodes mycelium polysaccharide to the colitis acute stage Colon and rectum effect length that DSS induces;
Figure 1B is the schematic diagram of Lentinula edodes mycelium polysaccharide on the colitis acute stage Colon and rectum cardinal principle pathological changes impact that DSS induces;
The schematic diagram of pathological changes impact under Fig. 1 C colitis acute stage colorectal carcinoma microscope that to be Lentinula edodes mycelium polysaccharide induce DSS;
Fig. 2 is the impact of Lentinula edodes mycelium polysaccharide on the colitis acute stage Colon and rectum length that DSS induces, #, compared with blank group, and P<0.05; *, compared with DSS model control group, P<0.05;
Fig. 3 colitis acute stage that to be Lentinula edodes mycelium polysaccharide induce DSS lesion score substantially impact, #, compared with blank group, P<0.05; *, compared with DSS model control group, P<0.05;
Fig. 4 is the impact of Lentinula edodes mycelium polysaccharide on the colitis acute stage lesion tissue scoring that DSS induces, #, compared with blank group, and P<0.05; *, compared with DSS model control group, P<0.05;
Fig. 5 is the schematic diagram of Lentinula edodes mycelium polysaccharide on the impact of colitis chronic phase Colon and rectum length, cardinal principle and lesion tissue that DSS induces;
Fig. 5 A is the schematic diagram of Lentinula edodes mycelium polysaccharide to the colitis chronic phase Colon and rectum effect length that DSS induces;
Fig. 5 B is the schematic diagram of Lentinula edodes mycelium polysaccharide on the colitis chronic phase Colon and rectum cardinal principle pathological changes impact that DSS induces;
The schematic diagram of pathological changes impact under Fig. 5 C colitis chronic phase colorectal carcinoma microscope that to be Lentinula edodes mycelium polysaccharide induce DSS;
Fig. 6 is the impact of Lentinula edodes mycelium polysaccharide on the colitis chronic phase Colon and rectum length that DSS induces, #, compared with blank group, and P<0.05; *, compared with DSS model control group, P<0.05;
Fig. 7 is the impact of Lentinula edodes mycelium polysaccharide on the colitis chronic phase Colon and rectum cardinal principle pathological changes that DSS induces, #, compared with blank group, and P<0.05; *, compared with DSS model control group, P<0.05;
Fig. 8 is the impact of Lentinula edodes mycelium polysaccharide on the colitis chronic phase colorectal carcinoma pathological changes that DSS induces, #, compared with blank group, and P<0.05; *, compared with DSS model control group, P<0.05;
Fig. 9 is the schematic diagram of Lentinula edodes mycelium polysaccharide on the Crohn disease hemafecia impact that TNBS induces;
Figure 10 is the impact of Lentinula edodes mycelium polysaccharide on the Crohn disease model mice body weight that TNBS induces, and *, compares with positive control drug SASP group with TNBS model group, P<0.05;
Figure 11 is the schematic diagram of Lentinula edodes mycelium polysaccharide to the Crohn disease Colon and rectum effect length that TNBS induces;
Figure 12 is the impact of Lentinula edodes mycelium polysaccharide on the Crohn disease Colon and rectum length that TNBS induces, #, and compared with 50% ethanol group of non-modeling, P<0.05, *, compare with positive control drug SASP group with TNBS model group, P<0.05;
Figure 13 is the impact of Lentinula edodes mycelium polysaccharide on the Crohn disease Colon and rectum cardinal principle pathological changes that TNBS induces; And
Figure 14 is the impact of Lentinula edodes mycelium polysaccharide on the Crohn disease colorectal carcinoma pathological changes that TNBS induces.
Detailed description of the invention
Below in conjunction with the accompanying drawing in the embodiment of the present invention, be clearly and completely described the technical scheme in the embodiment of the present invention, obviously, described embodiment is only the present invention's part embodiment, instead of whole embodiments.Based on the embodiment in the present invention, the every other embodiment that those of ordinary skill in the art obtain, all belongs to the scope of protection of the invention.
Embodiment 1 Lentinula edodes mycelium polysaccharide is to the therapeutical effect of the chronic ulcerative colitis acute stage that 5%DSS induces
Experiment material:
SPF level C57 mice 18 ~ 22g;
Dextran sulfate (DSS) is purchased from AMRESCO company;
Lentinula edodes mycelium polysaccharide, purchased from Kaifeng Pharmaceutical (Group) Co., Ltd., is lark powder, is provided by Kaifeng Pharmaceutical (Group) Co., Ltd., lot number: 20130914, total sugar content is 64.3%, and polyoses content is 60.0%, meets Chinese Pharmacopoeia version in 2010 two.
Experimental technique:
1. experimental mouse adapts to 7 days in standardization Animal House environment, gives standard animal feedstuff, drinks autoclaving water, and changes bedding and padding (weekly 2-3 time) in time.
2. configure 5%DSS solution: use normal saline according to w/v preparation 5%DSS solution, the DSS solution prepared is deposited in 4 DEG C of refrigerators.
3. after experiment starts, calculate by the amount of drinking water 6ml of every mice every day, each DSS solution pouring amount of drinking water every day in mice water bottle into gives mice and freely drinks (blank group mice drinks normal distilled water), supplements the DSS solution Summer Solstice or the Winter Solstice amount of drinking water next day.DSS solution gives 7 days altogether.
4. after last gives DSS24 hour, grouping gastric infusion is carried out to mice, be divided into 3 groups: blank group, model group and Lentinula edodes mycelium polysaccharide group, often organize 10.Blank group mice and model group mice give normal saline (NS) 0.1mL/ pcs/day every day, and Lentinula edodes mycelium polysaccharide group mice gives Lentinula edodes mycelium polysaccharide (20mg/kg), dosage period 7 days.
5. start modeling and record dietary amount every day of mice simultaneously, and weigh in, observe the general animation of mice and with or without hemafecia.
6. administration is plucked eyeball to every mice after 7 days and is got blood, the change of each inflammatory factor in test serum.Get the straight colon of mice and measure length, eye mesentery direction descending colon are longitudinally cut off, and cleaning feces, is fixed in 10% neutral formalin.
7. running water after fixing, carry out successively dewatering, paraffin embedding, section, HE dyeing, basis of microscopic observation pathological section is also marked.Standards of grading are as follows:
By 5% sodium dextran sulfate (DSS) inducing acute phase mouse models of colitis, observe the general state, defecation situation etc. of mice.Result shows, blank group mice is very active, and fur is smooth; Model control group mice lethargy, lazy dynamic, rough coat is matt, and Lentinula edodes mycelium polysaccharide group mice general state is good, and comparatively active, fur is smooth, and hair color is shinny, and appetite takes a turn for the better.Within the 3rd day after giving DSS, find, model control group mice has soft stool situation to occur, within the 4th day, starts to occur diarrhoea, and visual hemafecia.Lentinula edodes mycelium polysaccharide group mice bloody purulent stool and Mucous Stool disappear, stool molding.Administration is cut off after 7 days and is taken Colon and rectum and measure Colon and rectum length, visible lentinan group Colon and rectum length is obviously longer than model control group (with reference to Figure 1A, Fig. 2) cut open along mesentery stringer by Colon and rectum, perusal is respectively organized mouse intestinal general form and is changed.In DSS acute colitis animal model, based on distal colon pathological changes, can relate to total colectomy, show as mice pathological changes colon seriality edema, intestinal transmural is downright bad.Model group mice mucous hyperemia, edema, erosion and ulcer, ulcer surface is large, boundary clear, and inflammation is obvious, and part intestinal wall thickens, some animals colon and surrounding tissue adhesion (with reference to Figure 1B, Fig. 3).Lentinula edodes mycelium polysaccharide group murine colonic inflammation pathological changes is obviously lighter than model group, and mild inflammation is congested, edema, have no obviously rotten to the corn or ulcer (with reference to Figure 1B, Fig. 4), and group difference has significance (P<0.05).Get pathological changes colon site, fix through 10% neutral formalin, dewater, paraffin embedding, tissue slice, HE dyeing, tissues observed morphological change, and marking.Basis of microscopic observation mouse Colon mucosal tissue, normal group mouse Colon organizational structure rule, body of gland marshalling, a small amount of inflammatory cell infiltration.Model group mouse Colon mucosa suffers heavy damage, and part has defect, congested, edema in colon, and mucous epithelium has a large amount of neutrophilic granulocyte, lymphocyte and plasmocyte infiltrating as seen, based on neutrophil infiltration; Body of gland arrangement disorder, distortion in mucosa.Lentinula edodes mycelium polysaccharide group mice pathological changes alleviates, mucosa is complete, ulcer is all shown in healing, ulcer stove surface is covered by the epithelial cell of new life, inflammatory cell infiltration comparatively model group obviously reduces (with reference to figure 1C), score value is starkly lower than model control group, and difference has statistical significance (P<0.05, with reference to figure 4).
Be considered as curing with the tissue of Colon and rectum mucosa scoring <5 and calculate, Lentinula edodes mycelium polysaccharide is in the therapeutical effect of the chronic ulcerative colitis acute stage that 5%DSS induces, and cure rate is 43.8%.
Embodiment 2 Lentinula edodes mycelium polysaccharide is to the therapeutical effect of the chronic ulcerative colitis chronic phase that 1%DSS induces
Experiment material:
SPF level C57 mice 18 ~ 22g;
Dextran sulfate (DSS) is purchased from AMRESCO company;
Lentinula edodes mycelium polysaccharide, purchased from Kaifeng Pharmaceutical (Group) Co., Ltd., is lark powder, is provided by Kaifeng Pharmaceutical (Group) Co., Ltd., lot number: 20130914, total sugar content is 64.3%, and polyoses content is 60.0%, meets Chinese Pharmacopoeia version in 2010 two.
Experimental technique:
1. experimental mouse adapts to 7 days in standardization Animal House environment, gives standard animal feedstuff, drinks autoclaving water, and changes bedding and padding (weekly 2-3 time) in time.
2. configure 1%DSS solution: use normal saline according to w/v preparation 1%DSS solution, the DSS solution prepared is deposited in 4 DEG C of refrigerators.
3. after experiment starts, calculate by the amount of drinking water 6ml of every mice every day, each DSS solution pouring amount of drinking water every day in mice water bottle into gives mice and freely drinks (naive mice normally drinks distilled water), supplements the DSS solution Summer Solstice or the Winter Solstice amount of drinking water next day.DSS solution gives 20 days altogether.
4. after last gives DSS24 hour, grouping gastric infusion is carried out to mice, be divided into 3 groups: blank group, model control group and Lentinula edodes mycelium polysaccharide group, often organize 10 mices.Blank group mice gives normal saline (NS) 0.1mL/ pcs/day, and Lentinula edodes mycelium polysaccharide group mice gives 3mg/mL Lentinula edodes mycelium polysaccharide 0.1mL/ pcs/day, dosage period 20 days.
5. start modeling and should record dietary amount every day of mice simultaneously, and weigh in, observe the general animation of mice and with or without hemafecia.
6. administration is plucked eyeball to every mice after 20 days and is got blood, detects the change of each inflammatory factor in serum.Get the straight colon of mice and measure length, eye mesentery direction descending colon are longitudinally cut off, and cleaning feces, is fixed in 10% neutral formalin.
7. running water after fixing, carry out successively dewatering, paraffin embedding, section, HE dyeing, basis of microscopic observation pathological section is also marked.Standards of grading are as follows:
By 1% sodium dextran sulfate (DSS) inducing chronic phase mouse models of colitis, observe the general state, defecation situation etc. of mice.Result shows, blank group mice is very active, and fur is smooth; Model control group mice mobility is few, and rough coat is matt, and lentinan group mice general state is good, and comparatively active, fur is smooth, and hair color is shinny, and appetite takes a turn for the better.Within the 10th day after giving DSS, find, model control group mice has soft stool situation to occur, some animals visual hemafecia.Lentinula edodes mycelium polysaccharide group has no diarrhoea situation and occurs, stool molding.Within after modeling 20 days, take Colon and rectum and measure Colon and rectum length, visible lentinan group Colon and rectum length is obviously longer than model control group (with reference to figure 5A, Fig. 6).Cut open along mesentery stringer by Colon and rectum, perusal is respectively organized mouse intestinal general form and is changed.In the colitis chronic phase model that low concentration DSS induces, colonic pathological change is based on distal colon, total colectomy can be related to, model group mice mucous hyperemia, edema, erosion and ulcer, inflammation is obvious, part intestinal wall thickens, some animals colon and surrounding tissue adhesion (with reference to figure 5B, Fig. 7).Lentinula edodes mycelium polysaccharide group murine colonic inflammation pathological changes is obviously lighter than model group, mild inflammation is congested, edema, have no obviously rotten to the corn or ulcer (with reference to figure 5B, Fig. 7), and group difference has significance (P<0.05, with reference to figure 7).Get pathological changes colon site, fix through 10% neutral formalin, dewater, paraffin embedding, tissue slice, HE dyeing, tissues observed morphological change, and marking.Basis of microscopic observation mouse Colon mucosal tissue, normal group mouse Colon organizational structure rule, body of gland marshalling, a small amount of inflammatory cell infiltration.Model group mouse Colon mucosa injury, to come off, congested, edema in colon, the visible neutrophilic granulocyte of mucous epithelium, lymphocyte and plasmocyte infiltrating, based on neutrophil infiltration; Body of gland arrangement disorder, distortion in mucosa.Lentinula edodes mycelium polysaccharide group mice pathological changes alleviates, mucosa is complete, ulcer is all shown in healing, ulcer stove surface is covered by the epithelial cell of new life, inflammatory cell infiltration comparatively model group obviously reduces (with reference to figure 5C, Fig. 8), score value is starkly lower than model control group, and difference has statistical significance (P<0.05, with reference to figure 8).
Lentinula edodes mycelium polysaccharide is in the therapeutical effect of the chronic ulcerative colitis chronic phase that 1%DSS induces, and cure rate is 60%.
Embodiment 3 Lentinula edodes mycelium polysaccharide is to the therapeutical effect of the Crohn disease that TNBS induces
Experiment material:
SPF level C57 mice 18 ~ 22g, purchased from Beijing Military Medical Science Institute;
Trinitro-benzene-sulfonic acid (TNBS) is purchased from AMRESCO company;
Lentinula edodes mycelium polysaccharide, purchased from Kaifeng Pharmaceutical (Group) Co., Ltd., is lark powder, is provided by Kaifeng Pharmaceutical (Group) Co., Ltd., lot number: 20130914, total sugar content is 64.3%, and polyoses content is 60.0%, meets Chinese Pharmacopoeia version in 2010 two.
Experimental technique:
1. test C57 mice and adapt to 7 days in standardization Animal House environment, give standard animal feedstuff, drink autoclaving water, and change bedding and padding (weekly 2-3 time) in time.
2.C57 mice is divided into into blank group, 50% ethanol control group, trinitro-benzene-sulfonic acid (TNBS) model group, Lentinula edodes mycelium polysaccharide group, positive control drug SASP (sulfasalazine) group at random, often organizes 10.
The fasting of 3.C57 mice, can't help drink after 24 hours, etherization mice, 3.5F conduit inserts the dark about 5.5cm of intestinal from anus, every mice perfusion enema 100u1, blank group gives weekly normal saline 100ul coloclysis, and 50% ethanol control group gives 50% ethanol 100ul coloclysis.TNBS model group, positive control drug SASP group and Lentinula edodes mycelium polysaccharide group adopt 1 week TNBS2.0mg/50% ethanol enema induction C57 mice Crohn disease model, afterwards mice are inverted 60 seconds, then put back in cage.
4. after coloclysis 24h, grouping gastric infusion is carried out to mice: blank group, 50% ethanol control group and TNBS model group mice give normal saline 0.1mL/ pcs/day, lentinan group mice gives 3mg/mL Lentinula edodes mycelium polysaccharide 0.1mL/ pcs/day, positive control drug SASP group gives 40mg/mL sulfasalazine 0.1mL/ pcs/day, dosage period 7 days.
5. start modeling and should record dietary amount every day of mice simultaneously, and weigh in, observe the general animation of mice and with or without hemafecia.
6. administration is plucked eyeball to every mice after 7 days and is got blood, detects the change of each inflammatory factor in serum.Get the straight colon of mice and measure length, longitudinally being cut off by colon along mesentery direction, cleaning feces, is fixed in 10% neutral formalin.
7. running water after fixing, carry out successively dewatering, paraffin embedding, section, HE dyeing, basis of microscopic observation pathological section is also marked.Standards of grading are as follows:
Namely there is the situation such as hemafecia, loose stool, few dynamic, hair is matt, health is rolled up, appetite reduces, weight loss after 24 hours in the C57 mice modeling in TNBS colitis model group, blank group mice then reacts flexibly, appetite normally, body weight increase.1d after modeling, all there is diarrhoea in the rat of model group 70%.Lentinula edodes mycelium polysaccharide group diarrhea of mouse, hemafecia incidence rate and TNBS model group obviously reduce (with reference to figure 9), and Lentinula edodes mycelium polysaccharide group Mouse Weight comparatively model group obviously increase (with reference to Figure 10), mice general state is good, comparatively active.Sulfasalazine group mice is movable normal, and diarrhoea, hemafecia incidence rate obviously reduce than TNBS model group, but recovery extent is inferior to lentinan (with reference to figure 9).Administration takes Colon and rectum after 7 days and measures Colon and rectum length, visible sulfasalazine group and lentinan group Colon and rectum length are all obviously longer than model control group (with reference to Figure 11,12) mucous membrane of colon general form observes display (with reference to Figure 13), blank group mouse Colon has no rotten to the corn and ulcer, and colon wrinkle wall is neat; TNBS model group mouse Colon visible significantly ulcer companion is congested, edema, and notably occur pitchy slough, and intestinal wall thickens, the stenosis of far-end intestinal tube is narrow; The visible mouse Colon ulcer of Lentinula edodes mycelium polysaccharide group and necrosis area obviously reduce, and congested, edematous condition also obviously improves, and intestinal wall has no and obviously thickens.Model group mice substantially colon morphology is marked apparently higher than other groups (P<0.05).Mirror lower intestinal tract HE coloration result shows: the mucous layer of blank group (with reference to Figure 14) mouse Colon, tela submucosa and muscle layer and placenta percreta all complete, each Rotating fields of colon is clear; TNBS model group (with reference to Figure 14): visible intestinal mucosa epithelium damage, body of gland distortion, goblet cell lacks, at mucous layer and the visible a large amount of lymphocytic infiltration of tela submucosa; Lentinula edodes mycelium polysaccharide group mucous layer structural integrity, a small amount of glandular tube is misaligned, and goblet cell enriches, and has no inflammatory cell infiltration (with reference to Figure 14).
Lentinula edodes mycelium polysaccharide is used in the therapeutical effect of the Crohn disease of TNBS induction, and cure rate is 72.8%.
To sum up, Lentinula edodes mycelium polysaccharide obviously can alleviate Colon and rectum inflammation, the edema degree of chronic ulcerative colitis chronic phase, acute stage and Crohn disease mice, effectively can repair the mucosa injury of diseased mice, make ulcer healing, the obviously damage range of less diseased mice, and effectively can treat the diarrhoea of diseased mice.Visible, Lentinula edodes mycelium polysaccharide provided by the invention can treat ulcerative colitis and Crohn disease effectively.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, within the spirit and principles in the present invention all, any amendment done, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (7)
1. the application of Lentinula edodes mycelium polysaccharide in the medicine of preparation treatment inflammatory bowel.
2. application according to claim 1, is characterized in that, the application of described Lentinula edodes mycelium polysaccharide in the medicine of preparation treatment inflammatory bowel comprises: the application of described Lentinula edodes mycelium polysaccharide in the medicine preparing treatment ulcerative colitis and Crohn disease.
3. application according to claim 1 and 2, is characterized in that, the polyoses content in described Lentinula edodes mycelium polysaccharide is more than 60 % by weight.
4. application according to claim 3, is characterized in that, described Lentinula edodes mycelium polysaccharide is 0.83 ~ 3.32mg/kg/d to the dose therapeutically effective of people.
5. application according to claim 1, is characterized in that, the pharmaceutical pack of described treatment inflammatory bowel is containing Lentinula edodes mycelium polysaccharide, Lentinula edodes mycelium polysaccharide pharmaceutically acceptable salt, ester, hydrate or their combination and adjuvant.
6. application according to claim 5, it is characterized in that, the medicine of described treatment inflammatory bowel is selected from tablet, capsule, pill, suppository, aerosol, oral liquid, granule, powder, injection, syrup, medicated wine, tincture, distillate medicinal water, membrane or their combination.
7. application according to claim 5, is characterized in that, the administering mode of the medicine of described treatment inflammatory bowel comprises oral, injection, implantation, external, spraying, suction or their combination.
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| CN201410592300.3A Pending CN105535011A (en) | 2014-10-28 | 2014-10-28 | Applications of mushroom mycelium polysaccharides |
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| CN (1) | CN105535011A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108142923A (en) * | 2018-03-14 | 2018-06-12 | 广东省微生物研究所（广东省微生物分析检测中心） | Purposes of the mushroom dietary fiber extract in the preparation for preparing treatment and/or prevention intestinal bacilli illness relevant disease |
| CN110251515A (en) * | 2019-07-22 | 2019-09-20 | 天津国际生物医药联合研究院有限公司 | A kind of preparation method and application of Rho kinase inhibitor Y27632 load Lentinan compound |
| CN111329997A (en) * | 2020-03-04 | 2020-06-26 | 山东信得科技股份有限公司 | Soft chewable tablet for treating cat inflammatory intestinal diseases and preparation method thereof |
-
2014
- 2014-10-28 CN CN201410592300.3A patent/CN105535011A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| YOSUKE NISHITANI等: "Intestinal Anti-Inflammatory Activity of Lentinan: Influence on IL-8 and TNFR1 Expression in Intestinal Epithelial Cells", 《PLOS ONE》 * |
| 韩伟东等: "香菇多糖对溃疡性结肠炎大鼠肠道微生态失调的调整作用研究", 《中国微生态学杂志》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108142923A (en) * | 2018-03-14 | 2018-06-12 | 广东省微生物研究所（广东省微生物分析检测中心） | Purposes of the mushroom dietary fiber extract in the preparation for preparing treatment and/or prevention intestinal bacilli illness relevant disease |
| CN110251515A (en) * | 2019-07-22 | 2019-09-20 | 天津国际生物医药联合研究院有限公司 | A kind of preparation method and application of Rho kinase inhibitor Y27632 load Lentinan compound |
| CN111329997A (en) * | 2020-03-04 | 2020-06-26 | 山东信得科技股份有限公司 | Soft chewable tablet for treating cat inflammatory intestinal diseases and preparation method thereof |
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