CN107823211A - Application of the gucosamine in preparing ionising radiation and causing induced lung injury protective agents - Google Patents
Application of the gucosamine in preparing ionising radiation and causing induced lung injury protective agents Download PDFInfo
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- gucosamine
- ionising radiation
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
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Abstract
The present invention relates to new medicine use field, specifically application of the gucosamine in preparing ionising radiation and causing induced lung injury protective agents, the structural formula of the gucosamine are as follows.Gucosamine of the present invention causes induced lung injury protective agents as ionising radiation：Toxic side effect is small; it is evident in efficacy; the 3 days dosage with 100mg/kg/d concentration of pre-irradiation is administered; lung tissue can significantly be protected; suppress the conversion of lung tissue Epithelial and stromal; unique distinction of the gucosamine in protection ionising radiation causes induced lung injury is shown, medical domain has broad application prospects in China.
Description
Technical field
The present invention relates to new medicine use field, is that gucosamine is preparing ionising radiation cause lung radiation injury specifically
The application damaged in protective agents.
Background technology
Induced lung injury often betides breast tumor radiotherapy.Radiotherapy is the most important treatment of breast tumor
One of means, and it is then common tight of breast tumor radiotherapy that induced lung injury, which includes radiation pneumonitis and radiation fibrosis of lung,
Weight complication and dose limiting factor.The main clinical manifestation of induced lung injury is inflammatory infiltration, the progressive of alveolar spaces
Expiratory dyspnea, PFT deteriorate and ultimately result in respiratory failure.The generation of induced lung injury, is largely reduced
Receive the prognosis survival rate of chest irradiation patient.Therefore protection to the induced lung injury caused by breast tumor radiotherapy and control
Treatment has extremely important medical significance.At present clinically, in addition to using glucocorticoid symptomatic treatment, it there is no and effectively control
Treat medicine.There are not unified conclusion in the occurrence cause of induced lung injury, academia.It is most of to be considered due to ionising radiation
The cell factor immune imbalance of the inflammation cell of induction, fibroblast and its correlation, causes fibroblast excessively to increase
It is relevant with extracellular matrix (ECM) deposition to grow migration, so as to generate damage to body.At present, for induced lung injury still
Without effectively protection and medicine.Traditional radiation protection medicine is right while normal lung tissue plays radioprotective effect
Tumour cell also provides certain protection effect.Such medicine feature significantly reduces the effect of tumor radiotherapy, and this just makes
About its application in clinical tumor patient's chest radiotherapy.Therefore, in the clinical treatment of induced lung injury, urgently need
Find a kind of novel protective medicine evident in efficacy, toxic side effect is small, safe.Clinically mainly use sugared cortex at present
Hormone alleviates the acute inflammation stage reaction of induced lung injury, but the medicine is not alleviated for radiation fibrosis of lung at a specified future date
Effect, meanwhile, its serious side effect also limit scope and the dosage that the clinical drug uses.In animal experiment, radiation
Property protection medicine WR-2721, can effectively prevent and mitigate the acute inflammatory reaction of induced lung injury.But the medicine pair
Unobvious are acted in radiation fibrosis of lung at a specified future date and there is more strong bio-toxicity, limit its answering in clinic
With.In addition, the experimental study of induced lung injury protective agents, antioxidant, gene therapy, stem cell are concentrated mainly on
Several aspects such as treatment, but these methods are also respectively present the defects of effect is not notable, toxic side effect is big.Therefore, one is found
Kind is efficient, low toxicity, tumour cell is not protected even while radioactive protection effect is played to normal lung tissue
Possess the medicine of tumor-killing effect, be always urgent problem to be solved in induced lung injury research.
Gucosamine (Glucosamine) is widely present in nature, generally with N- acetyl derivatives (such as chitin)
Or it is present in microorganism, the polysaccharide of animal origin and combination in the form of N- sulfuric esters and N- acetyl -3-O- lactic acid ethers (muramic acid)
In polysaccharide.It is the compound that is substituted by an amino of a hydroxyl of glucose.Its molecular formula is C6H13O5N, it is commonly called as amino
Sugar, abbreviation ammonia sugar.It is the material of synthesis in human body, is the important nutrient to form cartilage cell, is healthy articular cartilage
Natural tissues composition.With advancing age, the shortage of the Glucosamine in human body is increasingly severe, and articular cartilage is constantly moved back
Change and wear.A large amount of medical researches in the U.S., Europe and Japan show：Glucosamine can help to repair and safeguard cartilage,
And the growth of cartilage cell can be stimulated.The daily health caring of human body is widely used at present.Gucosamine has anti-inflammatory, anti-oxidant
Effect, while find that its toxic side effect is very small, show extraordinary biological safety.Recent studies have found that glucose
Amine has significant lethal effect to a variety of human body tumour cells.With reference to the characteristics of gucosamine, the medicine is in clinical radiation
There is huge potential using value in injury of lungs.
The report in terms of induced lung injury preventing and treating on gucosamine is had no in the prior art.
The content of the invention
It is an object of the invention to provide the new application of gucosamine, i.e., causes induced lung injury to prevent in preparation ionising radiation
Control the application in medicine.
To achieve these goals, the first aspect of the present invention, there is provided gucosamine is preparing ionising radiation cause radioactivity
Application in injury of lungs protective agents, the structural formula of the gucosamine is as shown in following formula I：
Further, described gucosamine reduces the damage of lung tissue cell caused by ionising radiation.
Further, described gucosamine suppresses lung tissue inflammatory reaction caused by ionising radiation.
Further, described gucosamine suppresses the lung tissue Epithelial and stromal conversion of ionization radiation induction.
Further, it is excessive to suppress caused by ionizing radiation interstitial mark α-SMA and Vimentin for described gucosamine
Expression, and suppress caused by ionizing radiation interstitial mark E-cadherin downward.
Further, it is oral formulations that described ionising radiation, which causes induced lung injury protective agents,.
Further, the dosage of gucosamine is in described ionising radiation cause induced lung injury protective agents
100mg/kg/d, and 3 days before ionising radiation are administered.
Further, described ionising radiation is60Co gamma-rays irradiates.
The second aspect of the present invention, there is provided a kind of ionising radiation causes induced lung injury protective agents, and its active component is
Gucosamine.
Further, described ionising radiation causes induced lung injury protective agents also to include pharmaceutically acceptable auxiliary
Material.
The present invention by various concentrations (<Gucosamine 15mmol/L) is applied directly in normal pulmonary epithelial cells RLE-6TN,
Processing uses CCK-8 detection cell growths and propagation after 24 and 48 hours, as a result find：Gucosamine pair less than 10mmol/L
The growth of RLE-6TN cells and propagation do not produce inhibitory action.
Meanwhile the present invention using 5mmol/L gucosamines processing RLE-6TN cells after, then give cell various dose (0,
2nd, 4,8Gy)60Co gamma-rays is irradiated, and cloning efficiency detection cell growth and propagation, knot are used after then proceeding to culture 2 weeks
Fruit shows：, can be obvious with the increase of exposure dose after gucosamine (5mmol/L) pretreatment for the concentration that has no toxic side effect
Reduce the cell mortality for the RLE-6TN cells that radiotherapy is induced.Meanwhile the present invention is using at 5mmol/L gucosamines
After managing RLE-6TN cells, then cell 8Gy 60Co gamma-rays irradiation is given, then proceed to use after cultivating 24h
Hoechst33342 is dyed to detect Apoptosis, is as a result shown：The gucosamine (5mmol/L) for the concentration that has no toxic side effect
Pretreatment can obviously reduce the apoptosis rate for the RLE-6TN cells that radiotherapy is induced, and the gucosamine of concentration used
RLE-6TN is not had an impact.These datas show that gucosamine can reduce under without obvious toxic-side effects concentration
The damage of normal lung tissue's cell.
In addition, the female C57BL/6 mouse for 8 week old that the present invention obtains Chinese Academy of Sciences's Experimental Animal Center are used to move
Thing is tested.Mouse is fed in the cage of 25 ± 1 DEG C of daily replacing bedding and padding.Mouse is randomly divided into 4 groups：1st group, do not irradiate
+ saline control group；2nd group, irradiation+physiological saline group；3rd group, irradiation+WR2721 groups；4th group, irradiation+gucosamine
Group.3 days before the full lung of local irradiation, gucosamine (100mg/kg/d) and WR2721 (50mg/kg/d) pass through intraperitoneal injection to phase
The group answered.Then proceed to nursing mouse and mouse is put to death after 1 week, 8 weeks, take out lung tissue, fixed, wax embedding, carried out after section
HE is dyed.As a result show, it is normal not irradiate group alveolar wall construction.According to latter 1 week, simple irradiation group mouse was that visible alveolar septum increases
Thickness, and have substantial amounts of inflammatory cell infiltration.8 weeks after to photograph, it is seen that alveolar wall thickening aggravates, and alveolar collapses in various degree, into fibre
Dimension cell proliferation is obvious, and different degrees of fibrosis occurs in alveolar wall.Contrasted by irradiation group merely and gucosamine treatment group,
It is apparent that gucosamine lung tissue by according to early stage, that is, show good inhibition of inflammation.Meanwhile late
In the contrast of degree of inflammation, gucosamine treatment group has considerably lighter inflammatory reaction.In WR2721 treatment groups, mouse is shone
Lung tissue equally shows lighter inflammatory reaction.
Meanwhile the present invention will carry out α-SMA, Vimentin and E-cadherin SABCs after embedded block sections
Dyeing.As a result show, gucosamine suppresses caused by ionizing radiation interstitial mark α-SMA and Vimentin overexpression, and presses down
Caused by ionizing radiation interstitial mark E-cadherin downward is made.WR2721 treatment effects are markedly less than gucosamine group.
Show that lung tissue Epithelial and stromal conversion of the gucosamine for ionization radiation induction has significant inhibitory action.
Therefore, claimed gucosamine answering in preparing ionising radiation and causing induced lung injury protective agents
With.
Gucosamine provided by the present invention has following as preparation ionising radiation cause induced lung injury protective agents
Advantage：
1st, toxic side effect is small, is widely applied at present using gucosamine as arthritis field, document report Portugal
Grapes glucosamine has application as health treatment in the U.S., Japan and Europe；
2nd, evident in efficacy, the 3 days dosage with 100mg/kg/d concentration of pre-irradiation is administered, and can significantly protect lung tissue, presses down
Lung tissue Epithelial and stromal conversion processed.Above-mentioned performance shows gucosamine in protection ionising radiation causes induced lung injury
Unique distinction, in China, medical domain has broad application prospects.
Brief description of the drawings
Fig. 1 be embodiment 1 in various concentrations (<15 μm of ol/L) gucosamine processing 24 hours (A) and 48 hours (B) it is right
The influence of RLE-6TN cell growths and propagation；
Fig. 2 is that gucosamine pre-processes growth and propagation after RLE-6TN cells shine under different exposure doses in embodiment 2
Change；
Fig. 3 is that gucosamine pre-processes change of the RLE-6TN cells according to rear apoptosis in embodiment 3；
Fig. 4 is HE section contrasts after the full lung irradiation of gucosamine pretreatment of mice in embodiment 4.
Fig. 5 (A-D) is α-SMA, Vimentin and E- after the full lung irradiation of gucosamine pretreatment of mice in embodiment 5
The section contrast of cadherin immunohistochemical stainings.
Embodiment
Embodiment provided by the invention is elaborated with reference to embodiment.
Material：Cell line and cell culture：The normal pulmonary epithelial cells RLE-6TN of rat (American Cell collecting center) is existed
The cultures of RMPI 1640 containing 10% hyclone are based on 37 DEG C, 5%CO2Cultivated in incubator.Medicine and main agents：Medicine
Thing gucosamine, is dissolved in PBS, is made into mother liquor and is preserved for 1mol/L in 4 degree of refrigerators.The culture mediums of RMPI 1640, hyclone,
Pancreatin and WR2721 are purchased from Gibco companies；CCK-8 reagents are purchased from Japanese colleague's chemistry institute；Crystal violet, Hoechst dye liquors,
Propidium iodide (PI) is purchased from the green skies biotechnology research institute in Jiangsu.Mouse：8 weeks of Chinese Academy of Sciences's Experimental Animal Center acquisition
The female C57BL/6 mouse in age.
Embodiment 1：
(1) cell culture：By RLE-6TN cell culture in the RMPI culture mediums containing 10% hyclone.All cells
It is placed in 37 DEG C, 5%CO2Cultivated in incubator, per 2-3 days, once, growth period cell of taking the logarithm was used to test for passage.
(2) CCK-8 colorimetric methods：24h is by exponential phase cell (5*10 before drug-treated4/ mL) it is seeded to 96 orifice plates
In, every kind of concentration sets 6 parallel holes.The gucosamine for adding various concentrations continues culture 24 and 48 hours.In cell culture fluid
10 μ L CCK-8 reagents are added to continue to cultivate 1-4 hours.Absorbance at 450nm is determined using ELIASA.Finally using thin
The formula of born of the same parents' survival rate=dosing class value/control class value × 100% calculates cell survival rate.
Acquired results are as shown in Figure 1, the results showed that growth of the gucosamine to RLE-6TN cells less than 10mmol/L and
Propagation does not produce inhibitory action.The RLE-6TN cells LC50 (IC 50) of gucosamine processing in 48 hours is respectively
14.9mmol/L。
Embodiment 2：
(1) cell culture is the same as embodiment 1；
(2) cell clonal formation method：Take the logarithm growth period RLE-6TN cell, and be inoculated with by different exposure dose requirements
The cell of varying number (0,2,4,8Gy dose inoculation cell quantity is respectively 200,400,800 and 1600) into six orifice plates.
After 24 hours, gucosamine adds 5mmol/L gucosamines with radiation joint group and handled 1 hour, and control group is made using equivalent PBS
For negative control.Two groups of cells receive various dose (0-8Gy) simultaneously60Co gamma-rays irradiates, and continues culture 24 hours according to after,
Two groups change ordinary culture medium into, and continue until occurring terminating culture during obvious macroscopic cell clone in culture dish.
Culture medium is abandoned, PBS is washed twice, and fixes 30 minutes using absolute methanol, and Gimsa dye liquors dye 30 minutes, and flowing water dries in the air after rinsing
It is dry.Clone's colony more than 50 cells is counted under the microscope, and each stoichiometric point at least sets 3 Duplicate Samples, averaged, and calculates
Go out cell survival rate.Acquired results as shown in Fig. 2 the concentration that has no toxic side effect gucosamine (5mmol/L) pretreatment after, with
The increase of exposure dose, it can substantially reduce the cell mortality for the RLE-6TN cells that radiotherapy is induced.
Embodiment 3：
(1) cell culture is the same as embodiment 1；
(2) Hoechst33342 dyeing detection Apoptosis method：Take the logarithm growth period RLE-6TN cell (1*105/ mL) connect
Kind overnight, after adding the processing of 5mmol/L gucosamines, then gives cell 8Gy 60Co gamma-rays irradiation, then into six orifice plates
Continue after cultivating 24h, PBS is washed three times, and paraformaldehyde is fixed.Cell is detected after being dyed using Hoechst33342 and PI to wither
Die.Acquired results are as shown in figure 3, gucosamine (5mmol/L) pretreatment for the concentration that has no toxic side effect can obviously reduce radiation and control
The apoptosis rate of induced RLE-6TN cells is treated, and the gucosamine of concentration used does not have an impact to RLE-6TN.
Embodiment 4：
(1) mouse is fed：Mouse is placed in the cage of 25 ± 1 DEG C of daily replacing bedding and padding, ensures that moisture and food are sufficient.
(2) initially set up radiation and cause induced lung injury mouse model, from 6-8 week old female C57BL/6 mouse, at random
It is divided into three groups：Irradiation group (15Gy) 8 and pre-irradiation 3 days gucosamine administration groups 8, WR2721 administration groups 8 only reach control group 8
Only；With60Co gamma-rays carries out single chest irradiation to mouse, and mouse absorbed dose of radiation is 15Gy.By intragastric administration by glucose
Amine (100mg/kg/d) and WR2721 (50mg/kg/d) are delivered to respective sets.Every morning and observation at night after radioactive exposure
And record mouse.Put to death mouse within 1 week and 8 weeks after photograph respectively, take out lung tissue, fixed, wax embedding, HE dyes are carried out after section
Color.Acquired results such as Fig. 4 is shown, it is normal not irradiate group alveolar wall construction.According to latter 1 week, simple irradiation group mouse was visible alveolar
Septal thickening, and have substantial amounts of inflammatory cell infiltration.8 weeks after to photograph, it is seen that alveolar wall thickening aggravates, and alveolar collapses in various degree
Fall into, fibroblast increment is obvious, and different degrees of fibrosis occurs in alveolar wall.Handled by irradiation group merely and gucosamine
Group contrast, it is apparent that, gucosamine by according to early stage, that is, shows good inhibition of inflammation in lung tissue.Meanwhile
Late in the contrast of degree of inflammation, gucosamine treatment group has considerably lighter inflammatory reaction.It is small in WR2721 treatment groups
Mouse is equally shown lighter inflammatory reaction by according to lung tissue.
Embodiment 5：
(1) mouse is fed：Mouse is placed in the cage of 25 ± 1 DEG C of daily replacing bedding and padding, ensures that moisture and food are sufficient.
(2) radiation causes induced lung injury mouse model with embodiment 4.
(3) each group lung tissue wax stone of Example 4 carries out α-SMA, Vimentin and E-cadherin immunohistochemical stainings.
Section is taken pictures and counted.Acquired results such as Fig. 5 shows that gucosamine suppresses caused by ionizing radiation interstitial mark α-SMA
Over-expressed with Vimentin, and inhibit caused by ionizing radiation interstitial mark E-cadherin downward.WR2721 processing
Positive effect is weaker than gucosamine group.It is aobvious to show that lung tissue Epithelial and stromal conversion of the gucosamine for ionization radiation induction has
The inhibitory action of work.
Wherein, condition is irradiated：Radiation center (naval medicine institute of The 2nd Army Medical College, Chinese Shanghai)60Co gamma-rays shines
Penetrate.After being anaesthetized with 10% chloral hydrate (350mg/kg), full lung irradiation is carried out to mouse.All Radiatas receive single agent
15Gy, close rate 1Gy/min are measured, and is monitored after irradiation to 1 week.With 8Gy gamma-rays exposure dose rate 1Gy/min processing
Cell.
Statistical procedures：All experiments of above-described embodiment are repeated 3 times the above, as a result useRepresent.Using
SAS statistical softwares carry out t inspections to related data, with P<0.05 is to have significant difference.
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 (10)
1. application of the gucosamine in preparing ionising radiation and causing induced lung injury protective agents, the structure of the gucosamine
Formula is shown in formula I：
2. gucosamine according to claim 1 answering in preparing ionising radiation and causing induced lung injury protective agents
With, it is characterised in that described gucosamine reduces the damage of lung tissue cell caused by ionising radiation.
3. gucosamine according to claim 1 answering in preparing ionising radiation and causing induced lung injury protective agents
With, it is characterised in that described gucosamine suppresses lung tissue inflammatory reaction caused by ionising radiation.
4. gucosamine according to claim 1 answering in preparing ionising radiation and causing induced lung injury protective agents
With, it is characterised in that described gucosamine suppresses the lung tissue Epithelial and stromal conversion of ionization radiation induction.
5. gucosamine according to claim 4 answering in preparing ionising radiation and causing induced lung injury protective agents
With, it is characterised in that described gucosamine suppresses the excessive table of caused by ionizing radiation interstitial mark α-SMA and Vimentin
Reach, and suppress caused by ionizing radiation interstitial mark E-cadherin downward.
6. gucosamine according to claim 1 answering in preparing ionising radiation and causing induced lung injury protective agents
With, it is characterised in that it is oral formulations that described ionising radiation, which causes induced lung injury protective agents,.
7. gucosamine according to claim 1 answering in preparing ionising radiation and causing induced lung injury protective agents
With, it is characterised in that the dosage of gucosamine is in described ionising radiation cause induced lung injury protective agents
100mg/kg/d, and 3 days before ionising radiation are administered.
8. gucosamine according to claim 1 answering in preparing ionising radiation and causing induced lung injury protective agents
It is with, it is characterised in that described ionising radiation60Co gamma-rays irradiates.
9. a kind of ionising radiation causes induced lung injury protective agents, it is characterised in that described ionising radiation causes lung radiation injury
The active component for damaging protective agents is gucosamine.
10. ionising radiation according to claim 9 causes induced lung injury protective agents, it is characterised in that described electricity
Induced lung injury protective agents are caused also to include pharmaceutically acceptable auxiliary material from radiation.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109303764A (en) * | 2018-10-26 | 2019-02-05 | 山东贝诺医药生物科技有限公司 | A kind of chitosan cream and its preparation process preventing and treating radioactive skin reaction |
| CN111679071A (en) * | 2020-06-17 | 2020-09-18 | 南京医科大学 | Use of heme oxygenase-1 for diagnosis and treatment of radiation-induced lung injury |
| CN113476461A (en) * | 2021-08-23 | 2021-10-08 | 苏州大学 | Medicine for improving chemotaxis of inflammation-inhibiting type macrophages and application thereof |
| CN114948965A (en) * | 2022-07-18 | 2022-08-30 | 四川大学华西第二医院 | Application of compound in preparation of medicine for preventing and treating tissue injury |
| WO2023023994A1 (en) * | 2021-08-25 | 2023-03-02 | 苏州大学 | Drug for improving chemotaxis of anti-inflammatory macrophages and use thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109303764A (en) * | 2018-10-26 | 2019-02-05 | 山东贝诺医药生物科技有限公司 | A kind of chitosan cream and its preparation process preventing and treating radioactive skin reaction |
| CN111679071A (en) * | 2020-06-17 | 2020-09-18 | 南京医科大学 | Use of heme oxygenase-1 for diagnosis and treatment of radiation-induced lung injury |
| CN113476461A (en) * | 2021-08-23 | 2021-10-08 | 苏州大学 | Medicine for improving chemotaxis of inflammation-inhibiting type macrophages and application thereof |
| WO2023023994A1 (en) * | 2021-08-25 | 2023-03-02 | 苏州大学 | Drug for improving chemotaxis of anti-inflammatory macrophages and use thereof |
| CN114948965A (en) * | 2022-07-18 | 2022-08-30 | 四川大学华西第二医院 | Application of compound in preparation of medicine for preventing and treating tissue injury |
| CN114948965B (en) * | 2022-07-18 | 2023-11-10 | 四川大学华西第二医院 | Application of compound in preparation of medicine for preventing and treating tissue injury |
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