CN107296815A - Application of the zymosan in protection acute radiation bone marrow injury medicine is prepared - Google Patents
Application of the zymosan in protection acute radiation bone marrow injury medicine is prepared Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
- A61K31/736—Glucomannans or galactomannans, e.g. locust bean gum, guar gum
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Abstract
The present invention relates to biomedicine field, specifically application of the zymosan (Zymosan A) or derivatives thereof in protection acute radiation bone marrow injury medicine is prepared.The present invention passes through mouse experiment in vivo, it was demonstrated that zymosan can substantially mitigate the destructiveness of medulla hematopoietic system caused by ionising radiation, promotes the recovery of hemopoietic system, suppresses peripheral white blood cells caused by ionising radiation and declines；Tested by cell in vitro, zymosan can be reduced according to rear apoptosis rate, improved and shone rear cell proliferation activity；Prove that zymosan plays Study On The Radioprotective by targeted activation TLR2 signal paths by TLR2 knock-out mices.Illustrate to use zymosan to mitigate mouse bone marrow cells hemopoietic system radioactive damage by activating TLR2 signal paths according to preceding, protect peripheral white blood cells, new foundation is provided for zymosan protection acute radiation bone marrow injury.
Description
Technical field
It is that zymosan or derivatives thereof protects acute put in preparation specifically the present invention relates to biomedicine field
Application in penetrating property bone marrow injury medicine, and the pharmaceutical composition containing zymosan or derivatives thereof.
Background technology
At present, nuclear industry, nuclear energy and nuclear radiation tech are widely used for national economy, military field and medical field,
Core and the extensive use of radiation so that employee exposed to radiation and the public are greatly increased by the probability of nuclear radiation injury, nuclear radiation
Damage and protection are also more and more paid close attention to by educational circles and the public.After by heavy dose of ionising radiation, exposed personnel can
Generation acute radiation sickness, wherein acute radiation bone marrow injury (also known as bone marrow form acute radiation sickness) are high, difficult pre- because of its incidence
It is anti-and of increasing concern.
The mechanism of acute radiation bone marrow injury is mainly：The ionization of high-energy ray by directly act on and
The effect of connecing acts on intracellular biological macromolecular such as DNA, protein etc., causes medulla hematopoietic system bleeding, blood sinus destruction, marrow
Cell death, and then cause the clinical manifestations such as leukocyte count reduction, infection, bleeding.
The protection medicine for acute radiation bone marrow injury is mainly oxygen free radical scavenger (such as W2721) at present, is resisted
Scorching medicine (such as glucocorticoid), cytokine class (such as G-CSF).But these medicines are because secondary toxic action is larger, it is special to lack
Property, limit its extensive use in clinic.Therefore, find it is a kind of it is new effectively, toxic side effect is small, high specificity core spoke
Protection medicine is penetrated, is the emphasis and difficult point of current radioecology and radiation medicine research.
Zymosan (Zymosan-A, No. CAS：Yeast fungus cell membrane 58856-93-2) is extracted from, research has shown that it is
TLR2 (Toll-like receptor 2) part is (referring to document：Li JT,Wang WQ,Wang L,Liu NN,Zhao YL,
Zhu XS,Liu QQ,Gao CF,Yang AG,Jia LT:Subanesthetic isoflurane relieves
zymosan-induced neutrophil inflammatory response by targeting NMDA glutamate
receptor and Toll-like receptor 2signaling.Oncotarget 2016；7:31772-31789.).It is right
Body immune system can be activated by being concentrated mainly on it in the research of zymosan, strengthen immunity of organisms.Previously research is found,
The activation of TLR2 signal paths can promote NF- κ B nuclear translocations, raise cytokine profiles, play it to acute radiation marrow
The protection of damage is (referring to document：Gao F,Zhang C,Zhou C,Sun W,Liu X,Zhang P,Han J,Xian L,Bai
D,Liu H,Cheng Y,Li B,Cui J,Cai J,Liu C:A critical role of toll-like receptor
2(TLR2)and its'in vivo ligands in radio-resistance.Sci Rep 2015；5:13004.)
Not yet there is document report zymosan to play acute radiation bone marrow injury by activating TLR2 signal paths at present
The effect of protection.
The content of the invention
It is an object of the invention to provide a kind of new medical usage of zymosan or derivatives thereof, and contain yeast
The pharmaceutical composition of polysaccharide or derivatives thereof.
The first aspect of the present invention is preparing protection acute radiation bone marrow injury medicine there is provided zymosan or derivatives thereof
Application in thing.
Further, described protection acute radiation bone marrow injury medicine marrow hemopoiesis system caused by mitigation ionising radiation
System destructiveness, reduces and shines rear apoptosis rate, improves the medicine according to rear cell proliferation activity.Further, described mitigation
The medicine of medulla hematopoietic system destructiveness caused by ionising radiation destroys to reduce medulla hematopoietic system bleeding, bone marrow structure
Medicine.
Further, the medicine that described protection acute radiation bone marrow injury medicine is targeted activation TLR2.By preceding described
Understand, the activation of TLR2 signal paths can promote NF- κ B nuclear translocations, and up-regulation cytokine profiles play Study On The Radioprotective.
Zymosan and its derivative in the present invention refer to playing the zymosan compound of pharmacological action, are
The dextran polymer being formed by connecting by β -1,3 glycosidic bonds.
In order to verify protection effect of the zymosan for acute radiation bone marrow injury, and its whether pass through TLR2 signals
Path plays Study On The Radioprotective, and the present invention to two aspects of mouse experiment in vivo and cytological experiment in vitro by demonstrating
Zymosan is made to mouse bone marrow cells hemopoietic system and people's lymphocytic B cells AHH-1, people's intestinal epithelial cell HIEC radiation protection
With.
For experimental selection C57/BL6 in Mice Body, the male wild-type mice of 8 week old is tested, and pre-irradiation 24 is small
When and pre-irradiation give mouse peritoneal injection zymosan (25mg/kg/ times) within 2 hours.The radiation modality for protecting mouse is single
Full-body exposure, exposure dose is 7.0Gy, and close rate is 1Gy/min, and irradiation source is used60Co gamma-rays；To TLR2 knockout type mouse
Pre-irradiation 24 hours and pre-irradiation give mouse peritoneal injection zymosan (25mg/kg/ times) in 2 hours.Protect the irradiation of mouse
Mode is single full-body exposure, and exposure dose is 6.0Gy, and close rate is 1Gy/min, and irradiation source is used60Co gamma-rays；For thin
The experiment in vitro that born of the same parents learn, pre-irradiation 12 hours and shines preceding 2 hours, and 40ug/ml zymosan handles HIEC and AHH-1 cells,
The exposure dose gradient that the HIEC and AHH-1 cells of protection are received is 4.0Gy, 8.0Gy, and close rate 1Gy/min, irradiation source is adopted
With60Co gamma-rays.
It is found through experiments that, mouse can substantially mitigate Wild type mice bone marrow bleeding, blood sinus caused by irradiation according to preceding administration
Destruction, and bonemarrow nucleated cells number is protected, the apoptosis of bone marrow nucleated cell is reduced, and accelerate bone marrow injury reparation；Pass through TLR2
Knockout type mouse experiment confirms that zymosan plays Study On The Radioprotective by targeted activation TLR2；HIEC and AHH-1 is carried out
Test, irradiation energy induced apoptosis, suppression cell propagation, and can reduce cell according to preceding administration and shine rear apoptosis rate, improvement
Proliferation Ability situation.Illustrate there is core according to preceding can mitigate in chmice acute radioactivity bone marrow injury, protection marrow with zymosan
Cell, accelerates the recovery according to rear medulla hematopoietic system.
Further, described protection acute radiation bone marrow injury medicine is zymosan or derivatives thereof as unique living
Sexual element either includes the pharmaceutical composition of zymosan or derivatives thereof.
Any formulation can be made with the auxiliary material pharmaceutically commonly used in described medicine or pharmaceutical composition, and for example it can be
Decoction, powder, pill, vina, lozenge, jelly, plaster, medicinal tea, herbal leaven, cake agent, distillate medicinal water, stylus, line agent, medicated roll, nail agent, moxibustion
Press agent, paste, sublimed preparation, microencapsulation, vein emulsion, Liposomal formulation, aerosol, precursor medicine preparation, injection, mixture, mouth
Ampulla, tablet, capsule, pill, emulsion, ointment, rubber plaster, film, sponginum, ionophore are taken, or thoroughly
Skin absorbent.
The second aspect of the present invention is more by yeast there is provided a kind of pharmaceutical composition of protection acute radiation bone marrow injury
Sugar or derivatives thereof and pharmaceutically acceptable auxiliary material composition.
Zymosan of the present invention or derivatives thereof is used to prevent acute radiation bone marrow injury, is particularly suitable for use in atomic warfare
Strive, nuclear accident, employee exposed to radiation, tumor radiotherapy treatment patient etc. with acute radiation bone marrow injury.The ferment of the present invention
Female polysaccharide or derivatives thereof is used especially for preventing acute radiation bone marrow injury, i.e., radioactivity bone may be met with by delivering medicine in advance
Marrow damage crowd, administering mode be not limited to orally, inject etc..
Beneficial effect of the present invention is：
The present invention is by mouse experiment in vivo, and zymosan can substantially mitigate medulla hematopoietic system caused by ionising radiation
Destructiveness, promote hemopoietic system recovery, suppress ionising radiation caused by peripheral white blood cells decline；Pass through cell in vitro
Experiment is learned, zymosan can reduce according to rear apoptosis rate, improve and shine rear cell proliferation activity；Pass through TLR2 knockout type mouse
Confirm that zymosan plays Study On The Radioprotective by targeted activation TLR2；Tested by cell in vitro, zymosan can
Reduce cell and shine rear apoptosis rate, improve Proliferation Ability situation, illustrating can be by targeted activation TLR2 with zymosan according to before
Mitigate chmice acute radioactivity bone marrow injury, protect bone marrow nucleated cell, protect peripheral white blood cells, reduce mouse bone marrow cells hematopoiesis
Sensitiveness of the system to radiation.Therefore, the present invention provides for zymosan or derivatives thereof protection acute radiation bone marrow injury
New foundation.
The effects such as research at present, which has proven to zymosan, has absorption pathogen, antitumor and enhancing immunologic function, and
The invention provides the new indication of zymosan, the damage brought for protection nuclear radiation provides new medicine.
Brief description of the drawings
Fig. 1 is the influence result figure of bone marrow injury after zymosan irradiates to mouse.Wherein, A is that zymosan mitigation is small
The result figure of bone marrow injury after mouse irradiation, B is the result figure of mouse bone marrow cells karyocyte, the situation for weighing bone marrow injury.
Fig. 2 is influence result figure of the zymosan to mouse peripheral blood leucocyte caused by irradiation.
Fig. 3 is influence result figure of the zymosan to mice spleen coefficient (spleen weight/body weight).
Fig. 4 is influence result figure of the zymosan to cell in vitro AHH-1 apoptosis.Wherein, A is AnnexinV/PI reagents
The result that box passes through flow cytomery；B is the quantitative statisticses result figure that zymosan influences on apoptosis rate.
Fig. 5 is the result figure that zymosan mitigates HIEC inhibited proliferations.
Fig. 6 is that zymosan influences result figure to the life cycle of TLR2 wild types and TLR2 knockout type mouse.
Embodiment
The embodiment provided with reference to embodiment and accompanying drawing the present invention elaborates.
Embodiment 1：Zymosan can mitigate bone marrow injury after mouse irradiation
First, experimental method
Choose C57/BL6, the male mice of 8 week old to be tested, mouse is divided into zymosan group and control group two
Group.Pre-irradiation 24 hours and pre-irradiation give mouse peritoneal injection zymosan (25mg/kg/ times) in 2 hours, because zymosan is used
Sterile saline is dissolved, therefore the sterile saline of same dose is injected intraperitoneally in control group mice.
Zymosan group and control group mice irradiation use single full-body exposure, and dosage is 7.0Gy, and close rate is 1Gy/
Min, irradiation source is used60Co gamma-rays.
Mouse bone marrow cells were won respectively at the 1st, 5,10,15,30 day carry out pathological examination after irradiation.Pathological examination contaminates for HE
Color, display myeloid tissue structure and lesion situation.Except pathological examination, we also measure frangments smear, periphery
Leukocyte Counts and spleen index, spleen index are to calculate the spleen tissue and body weight ratio newly taken out, are indicated by the ratio
The degree of impairment of peripheral blood system, so as to speculate the degree of injury of medulla hematopoietic system indirectly.General hemopoietic system damage is got over
Seriously, spleen index is lower.
2nd, experimental result
Above experimental result is shown by Fig. 1-Fig. 3：
(1) Sections of Bone Marrow HE dyeing indicates the bone marrow of mice difference of the 1st day control group and administration group after irradiation not
Substantially；The 5th day and the 10th day after irradiation, control group is obvious compared with administration group damage to be aggravated, and bonemarrow nucleated cells number is significantly reduced；
According to latter 20th day, hemopoietic system started to recover, and administration group is recovered to accelerate (Figure 1A) compared with control group.Pass through flow cytometer detection mouse bone marrow cells
Number of nucleated cells also points out administration group substantially to mitigate (Figure 1B) compared with control group damage.Therefore, zymosan alleviates mouse irradiation
The situation of bone marrow injury afterwards.
(2) leukocyte counts are shown, administration group significantly improves peripheral white blood cell (Fig. 2).
(3) spleen coefficient results are shown, irradiation causes spleen coefficient to lower, and zymosan can increase spleen coefficient, illustrate ferment
Female polysaccharide has played hemopoietic system protective effect (Fig. 3).
Embodiment 2：Zymosan mitigates the apoptosis after AHH-1 shines
First, experimental method：
In order to verify influence of the zymosan to Apoptosis after irradiation, Apoptosis inspection is carried out to the AHH-1 after irradiation
Survey.
Apoptosis detection is using the double dyes of AnnexinV/PI (being purchased from Invitrogen companies) flow cytometer detection method, its principle
It is：Apoptosis is the process of a procedural startup, can be divided into early apoptosis, two processes of late apoptic.In early apoptosis
When, phosphatidylserine (PS) can be changed into turning up from the state originally on the inside of cellular phospholipid film, now Annexin V can with it is outer
The PS turned over is combined, and is indicated as a detection of early apoptosis.Cell late apoptosis stage, permeability of cell membrane increase, this
When propidium iodide (Propidium Iodide, PI) can then be combined as a kind of nucleic acid dye into cell interior with nucleic acid, carefully
Karyon is dyed to red.Fluorescein on Annexin V marks, with PI conjunctive uses just can by the analysis of flow cytometer come
Identify Level of Apoptosis (including early apoptosis and late apoptic).
Concrete operation step is：Ferment was given in 2 hours in pre-irradiation 12 hours and pre-irradiation to AHH-1 (being purchased from ATCC) cell
Female polysaccharide (40ug/ml) or sterile saline, are then irradiated to cell, and dosage is 4.0Gy and 8.0Gy, close rate
For 1Gy/min.Vitellophag and carry out carrying out fluidic cell point after Annexin V/PI dyeing, 20min after 24 hours after irradiation
Analysis.
2nd, experimental result
As shown in figure 4, irradiation+zymosan group compared with irradiation+NS group apoptosis rate substantially reduce, show as upper right and bottom right as
Limit cell proportion and reduce (Fig. 4 A).Apoptosis rate is that coordinate diagram right upper quadrant adds right lower quadrant cell proportion sum, and statistics is fixed
Amount display, zymosan substantially reduces cell according to rear apoptosis rate (Fig. 4 B).
Embodiment 3：Zymosan mitigates inhibited proliferation of the irradiation to HIEC
First, experimental method：
The detection of ability of cell proliferation is tested using CCK-8, and its principle is：WST -8 in CCK-8 reagents can be by cell
Dehydrogenase in mitochondria is reduced to the yellow formazan product with high water soluble, the quantity and living cells of generation formazan things
Quantity be directly proportional.Its absorbance value is determined at 450nm wavelength with enzyme-linked immunosorbent assay instrument, viable count can be reflected indirectly
Amount.
Concrete operation step is：Pre-irradiation 24 hours, by HIEC plating cells in 96 orifice plates (5000/hole), treats cell
After adherent, agent-feeding treatment is carried out according to first 12 hours and according to first 2 hours, gives zymosan (40ug/ml) and NS.Exposure dose point
Not Wei 8.0Gy, close rate is 1Gy/min, according to after change culture medium continue cultivate, cell was examined with enzyme linked immunological in 24 hours according to after
Survey instrument and its absorbance value is determined at 450nm wavelength.
2nd, experimental result
As shown in figure 5, compared to control, administration group cell absorbance substantially increases, illustrate that zymosan mitigates irradiation to thin
The inhibitory action of born of the same parents, promotes the proliferation activity according to rear cell.
Embodiment 4：TLR2 knocks out the Study On The Radioprotective for having reversed zymosan to mouse
First, test method
Choose TLR2 knockouts type, the male mice of 8 week old to be tested, mouse is divided into zymosan group and control group
Two groups.Pre-irradiation 24 hours and pre-irradiation give mouse peritoneal injection zymosan (25mg/kg/ times) in 2 hours, because of zymosan
Dissolved with sterile saline, therefore the sterile saline of same dose is injected intraperitoneally in control group mice.
Zymosan group and control group mice irradiation use single full-body exposure, and dosage is 6.0Gy, and close rate is 1Gy/
Min, irradiation source is used60Co gamma-rays.
Mouse mean survival time and life cycle are observed after irradiation.
2nd, experimental result
As shown in fig. 6, administration group is compared with control group, life cycle and no significant difference, zymosan to TLR2 knock-out mices simultaneously
Radiationless protective action, it was demonstrated that zymosan plays Study On The Radioprotective and mainly passes through TLR2 signal paths.
Above-described embodiment explores Acute Meyloid damage after zymosan irradiates to mouse, to AHH- by taking zymosan as an example
1 acts on according to rear apoptosis and to the mitigation of HIEC Proliferation Ability, but the invention is not restricted to zymosan, can play same medicine
The zymosan derivative of reason effect falls within protection scope of the present invention.
The preferred embodiment to the invention is illustrated above, but the invention be not limited to it is described
Embodiment, those skilled in the art can also make a variety of equivalent on the premise of without prejudice to the invention spirit
Modification or replacement, these equivalent modifications or replacement are all contained in the application claim limited range.
Claims (7)
1. application of the zymosan or derivatives thereof in protection acute radiation bone marrow injury medicine is prepared.
2. zymosan according to claim 1 or derivatives thereof is in protection acute radiation bone marrow injury medicine is prepared
Application, it is characterised in that described protection acute radiation bone marrow injury medicine for mitigate ionising radiation caused by marrow make
Blood system destruction degree, reduces and shines rear apoptosis rate, improves the medicine according to rear cell proliferation activity.
3. zymosan according to claim 2 or derivatives thereof is in protection acute radiation bone marrow injury medicine is prepared
Application, it is characterised in that the described medicine for mitigating medulla hematopoietic system destructiveness caused by ionising radiation is reduces bone
The bleeding of marrow hemopoietic system, the medicine of bone marrow structure destruction.
4. zymosan according to claim 1 or derivatives thereof is in protection acute radiation bone marrow injury medicine is prepared
Application, it is characterised in that the medicine that described protection acute radiation bone marrow injury medicine is targeted activation TLR2.
5. protection acute radiation bone marrow injury is being prepared according to any described zymosans of claim 1-4 or derivatives thereof
Application in medicine, it is characterised in that described protection acute radiation bone marrow injury medicine is zymosan or derivatives thereof
The pharmaceutical composition of zymosan or derivatives thereof is either included as sole active composition.
6. zymosan according to claim 1 or derivatives thereof is in protection acute radiation bone marrow injury medicine is prepared
Application, it is characterised in that described protection acute radiation bone marrow injury medicine be decoction, powder, pill, vina, lozenge,
Agent is pressed in jelly, plaster, medicinal tea, herbal leaven, cake agent, distillate medicinal water, stylus, line agent, medicated roll, nail agent, moxibustion, paste, sublimed preparation, microencapsulation,
Vein emulsion, Liposomal formulation, aerosol, precursor medicine preparation, injection, mixture, peroral ampule agent, tablet, capsule, dripping pill
Agent, emulsion, ointment, rubber plaster, film, sponginum, ionophore, or cutaneous permeable agent.
7. a kind of pharmaceutical composition of protection acute radiation bone marrow injury, it is characterised in that by zymosan or derivatives thereof
And pharmaceutically acceptable auxiliary material composition.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110075269A (en) * | 2019-04-19 | 2019-08-02 | 中国人民解放军第二军医大学 | Murabutide causes to apply in marrow, small intestine and splenic injury protective agents in preparation ionising radiation |
| CN114732912A (en) * | 2022-05-06 | 2022-07-12 | 中国人民解放军海军军医大学 | Application of Zym-Dep in preparation of anti-radiation medicine or medicine for treating ionizing radiation injury |
| CN114886912A (en) * | 2022-03-29 | 2022-08-12 | 中国人民解放军海军军医大学 | Application of Zymosan-A in intestinal radiation injury protection |
| EP3886916A4 (en) * | 2018-11-26 | 2023-03-29 | Duke University | Compositions and methods for inducing scarring by peri-tumoral cells |
| CN116370495A (en) * | 2023-05-05 | 2023-07-04 | 中国人民解放军海军军医大学第一附属医院 | Application of Zymosan-A in preparation of ovarian anti-radiation drugs or drugs for treating ovarian ionizing radiation injury |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP3886916A4 (en) * | 2018-11-26 | 2023-03-29 | Duke University | Compositions and methods for inducing scarring by peri-tumoral cells |
| US11998614B2 (en) | 2018-11-26 | 2024-06-04 | Duke University | Compositions and methods for inducing scarring by peri-tumoral cells |
| CN110075269A (en) * | 2019-04-19 | 2019-08-02 | 中国人民解放军第二军医大学 | Murabutide causes to apply in marrow, small intestine and splenic injury protective agents in preparation ionising radiation |
| CN110075269B (en) * | 2019-04-19 | 2021-06-29 | 中国人民解放军第二军医大学 | Application of Murabutide in preparation of medicine for preventing and treating bone marrow, small intestine and spleen injuries caused by ionizing radiation |
| CN114886912A (en) * | 2022-03-29 | 2022-08-12 | 中国人民解放军海军军医大学 | Application of Zymosan-A in intestinal radiation injury protection |
| CN114732912A (en) * | 2022-05-06 | 2022-07-12 | 中国人民解放军海军军医大学 | Application of Zym-Dep in preparation of anti-radiation medicine or medicine for treating ionizing radiation injury |
| CN116370495A (en) * | 2023-05-05 | 2023-07-04 | 中国人民解放军海军军医大学第一附属医院 | Application of Zymosan-A in preparation of ovarian anti-radiation drugs or drugs for treating ovarian ionizing radiation injury |
| CN116370495B (en) * | 2023-05-05 | 2024-08-27 | 中国人民解放军海军军医大学第一附属医院 | Use of Zymosan-A in preparation of ovary anti-radiation medicine or medicine for treating ovary ionizing radiation injury |
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Application publication date: 20171027 |