CN109771637A - A kind of albumen of anti-atopic dermatitis - Google Patents
A kind of albumen of anti-atopic dermatitis Download PDFInfo
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- CN109771637A CN109771637A CN201910089815.4A CN201910089815A CN109771637A CN 109771637 A CN109771637 A CN 109771637A CN 201910089815 A CN201910089815 A CN 201910089815A CN 109771637 A CN109771637 A CN 109771637A
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- atopic dermatitis
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Abstract
The present invention provides a kind of purposes of the anti-atopic dermatitis of albumen, also disclose the pharmaceutical composition of corresponding anti-atopic dermatitis, can contain pharmaceutically acceptable carrier, and dosage form can be injection.Contribution of the invention is, provides a kind of completely new anti-atopic dermatitis medicine.
Description
Technical field
The invention belongs to field of biotechnology.
Background technique
Atopic dermatitis (atopic dermatitis, abbreviation AD) is a kind of chronic skin disease, it is characterised in that dry
It is dry, the selectivity accumulation of itch and erythematous eczema and inflammatory cell.The AD state of an illness easily repeatedly, is not easy to cure, and leads to the body of patient
Body Health and Living quality is all severely impacted.The morbidity of AD be gene genetic, environmental factor, skin barrier function defect and
Up to the present the coefficient result of many aspects such as dysimmunity not yet illustrates its pathogenesis completely.In recent years, AD
Disease incidence increase year by year, the treatment of AD becomes the important topic that is concerned.The treatment of current AD uses antihistamine and class more
The hormone medicines such as steroid hormone, long-time service can generate the side effects such as drug resistance and atrophoderma.
The virulence factor of helicobacter pylori has helicobacter pylori-neutrophil activating protein (Helicobacter
Pylori neutrophil-activating protein, Hp-NAP), CagA, CagPAI, VacA, OipA, BabA etc.,
It can cause inflammatory reaction, study the important virulence factor that discovery Hp-NAP is helicobacter pylori through this seminar.This laboratory exists
First, China has submitted the coding gene sequence (Genebank accession number AY366361) of Hp-NAP, also utilizes genetic engineering skill
For art by its clonal expression, affinity chromatography obtains helicobacter pylori-neutrophil activating protein (Hp-NAP).Currently, Hp-NAP
Treatment of atopic dermatitis is acted on there is not yet correlative study.
Summary of the invention
The invention discloses Hp-NAP helicobacter pylori-neutrophil activating protein anti-atopic dermatitis purposes,
Sequence can inquire accession number AY366361 in Genebank and obtain；The present invention also correspondence discloses the anti-idiocrasy skin containing the albumen
Scorching pharmaceutical composition, the pharmaceutical composition contain pharmaceutically acceptable carrier, and dosage form can be injection, such as powder-injection or injection
Liquid, administration mode can be intraperitoneal injection.
The applicant has found that Hp-NAP can effectively treat AD, and have studied its effect by the AD model that oxazolone induces
Mechanism provides a kind of completely new drug for AD therapy field.
Detailed description of the invention
Figure 1A is the analysis that each experimental group influences AD mouse ear thickness；
Figure 1B is the analysis that each experimental group takes pictures to AD mouse systemic and ear, and arrow indicates the amplification to ear part；
Fig. 2A is the statistical analysis of each experimental mice otic tissues skin layer thickness；
Fig. 2 B is the statistical analysis of each experimental mice ear Number of Mast cells；
The unified meaning for the diagram that above each figure may relate to: * indicates that p < 0.05, * * indicate that p < 0.01, * * * indicate P <
0.001。
Specific embodiment
Therapeutic effect experimental method of the Hp-NAP albumen to the OXA atopic dermatitis model mouse induced:
(1) AD mouse model is administered
A. the BALB/C mice for buying 7 week old female, laboratory rearing after a week, mouse is randomly divided into 4 groups ---
Blank control group, sensitization group, HP-NAP protein medicine-feeding group, dexamethasone administration group (every group each 6), are placed on independent ventilation cage box
Middle raising, and mark.
B. by the BALB/C mice back shaving about 2cm X 2cm area of 8 week old, with 5% oxazolone after 24 hours
(Oxazolone) 20 μ L of sensitization liquid smears sensitization in BALB/c mouse back, and 20 μ L acetone are smeared at blank control group mouse back
Olive oil solution.
C. after a week, 0.3% 20 μ L of oxazolone solution is applied on the inside of mouse ear, excites (blank pair for ear
According to group with acetone: olive oil 4:1 is replaced), three-times-weekly.
D.HP-NAP protein medicine-feeding scheme: beginning intraperitoneal administration from the 0th day totally 7 times, is given every time 3 times a week after modeling
Medicine 200ug/0.2ml.At 0-14 days, after each ear excites one hour, each group mouse is administered respectively: sensitization group is infused in abdominal cavity
Penetrate 200 μ L PBS solutions；HP-NAP protein medicine-feeding group is in 200 μ g/0.2mL HP-NAP protein solutions of intraperitoneal injection；Dexamethasone
Administration group is in 200 μ g/0.2mL dexamethasone (DEX) solution of intraperitoneal injection.It is administered three-times-weekly, the 16th day sacrifice mouse.
(2) AD mouse the skin being grievously injured degree
At -7-16 days, Mice Auricle part thickness is measured with feeler daily, measurement is completed before sensitization or excitation,
Each experimental mice skin lesion situation of observation AD model is simultaneously taken pictures.
(3) the Histopathology detection of AD mouse
A. at the mouse invasion of AD model the 16th day, anesthetic sacrifices mouse, cuts the ear of BALB/c mouse morbidity
Tissue, and place it in 4% paraformaldehyde fixer and fix more than for 24 hours.
B. after carrying out paraffin embedding processing to mouse ear tissue, 6 μm of slices are cut into, otic tissues carry out
H&E dyeing after H&E slice is taken pictures, and analyzes photo using ImageJ software, counts ear's epidermal thickness.
C. after carrying out paraffin embedding processing to mouse ear tissue, 6 μm of slices are cut into, otic tissues carry out
Toluidine blue staining after slice is taken pictures, and analyzes photo using ImageJ software, counts the infiltration of inflammatory cell in otic tissues
Situation.
Experimental result:
1, improvement of the HP-NAP albumen to atopic dermatitis mouse symptom
Sensitization group: since the 7th day, mouse ear swelling degree was obviously increased, and ear's redness degree is serious, until the
14-16 days, incrustation is gradually formed, lichenification occurs.
HP-NAP treatment group: at 14-16 days, compared with sensitization group, the atopic dermatitis ear of mouse did not formed a scab
And red swollen phenomenon is obviously inhibited, ear thickness mitigates.
Wherein, Figure 1A is shown in the data statistics of ear thickness, and ear photo is as shown in Figure 1B before sacrificing mouse, it is clear that HP-NAP albumen
Group has notable difference compared with sensitization group.
2, the Histopathology detection of AD mouse
H&E staining analysis shows that sensitization group keratoderma occurs and destroyed, and skin corium thickens, a large amount of inflammatory cells
The phenomenon that infiltration, blood vessel dilatation；And there is no the exudation of apparent inflammatory cell in HP-NAP administration group, epidermis increases in slight
Thickness, cuticula is complete, and blood vessel is more significant (being detailed in Fig. 2A) without obvious expansion, i.e. HP-NAP protein medicine-feeding group therapeutic effect.
Toluidine blue staining is carried out to otic tissues, observes mast cells infiltration situation in sensitization group otic tissues.As a result
Show that sensitization group ear epidermis nearby has and is largely dyed to purple or purplish red coloured particles, i.e. mast cell.And HP-NAP is administered
Group ear's Toluidine blue staining analysis shows, nearby Mast Cells are reduced ear's epidermis, and the phenomenon that mast cells infiltration subtracts
Gently (it is detailed in Fig. 2 B).
Claims (5)
1. the purposes that albumen Hp-NAP is used to prepare anti-atopic dermatitis drug.
2. the pharmaceutical composition of anti-atopic dermatitis, which includes albumen Hp-NAP and pharmaceutically acceptable load
Body.
3. pharmaceutical composition as claimed in claim 2, characterized in that the dosage form of described pharmaceutical composition is injection.
4. pharmaceutical composition as claimed in claim 2 or claim 3, characterized in that the dosage form of described pharmaceutical composition be powder-injection or
Injection.
5. the pharmaceutical composition as described in any first claim, characterized in that the administration mode of described pharmaceutical composition is
Intraperitoneal injection.
Priority Applications (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910089815.4A CN109771637B (en) | 2019-01-30 | 2019-01-30 | Protein for resisting atopic dermatitis |
| US17/427,109 US20220096595A1 (en) | 2019-01-30 | 2020-03-30 | Anti-atopic dermatitis protein |
| PCT/CN2020/082001 WO2020156593A1 (en) | 2019-01-30 | 2020-03-30 | Anti-atopic dermatitis protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910089815.4A CN109771637B (en) | 2019-01-30 | 2019-01-30 | Protein for resisting atopic dermatitis |
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| Publication Number | Publication Date |
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| CN109771637A true CN109771637A (en) | 2019-05-21 |
| CN109771637B CN109771637B (en) | 2019-12-06 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201910089815.4A Active CN109771637B (en) | 2019-01-30 | 2019-01-30 | Protein for resisting atopic dermatitis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020156593A1 (en) * | 2019-01-30 | 2020-08-06 | 郑州大学 | Anti-atopic dermatitis protein |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102002106A (en) * | 2010-05-20 | 2011-04-06 | 郑州大学 | Fusion protein MBP-NAP and preparation method and application thereof |
| CN103558386A (en) * | 2013-08-22 | 2014-02-05 | 郑州大学 | Method for detecting rMBP-NAP of fusion protein through ELISA |
| CN103923935A (en) * | 2014-04-22 | 2014-07-16 | 郑州大学 | sIL-4R-NAP (soluble interleukin-4 receptor-neutrophil-activating protein) fusion gene |
| CN107875375A (en) * | 2017-10-10 | 2018-04-06 | 郑州大学 | RMBP NAP regulate and control macrophage and direct anti-tumor application |
| CN108728473A (en) * | 2017-11-30 | 2018-11-02 | 新乡医学院 | A kind of expression recombinant vector of helicobacter pylori NapA albumen, recombinant bacterial strain and preparation method thereof, application |
-
2019
- 2019-01-30 CN CN201910089815.4A patent/CN109771637B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102002106A (en) * | 2010-05-20 | 2011-04-06 | 郑州大学 | Fusion protein MBP-NAP and preparation method and application thereof |
| CN103558386A (en) * | 2013-08-22 | 2014-02-05 | 郑州大学 | Method for detecting rMBP-NAP of fusion protein through ELISA |
| CN103923935A (en) * | 2014-04-22 | 2014-07-16 | 郑州大学 | sIL-4R-NAP (soluble interleukin-4 receptor-neutrophil-activating protein) fusion gene |
| CN107875375A (en) * | 2017-10-10 | 2018-04-06 | 郑州大学 | RMBP NAP regulate and control macrophage and direct anti-tumor application |
| CN108728473A (en) * | 2017-11-30 | 2018-11-02 | 新乡医学院 | A kind of expression recombinant vector of helicobacter pylori NapA albumen, recombinant bacterial strain and preparation method thereof, application |
Non-Patent Citations (3)
| Title |
|---|
| D"ELIOS MM等: "The neutrophil-activating protein of Helicobacter pylori (HP-NAP) as an immune modulating agent", 《FEMS IMMUNOL MED MICROBIOL》 * |
| HATANO等: "Murine atopic dermatitis responds to peroxisome proliferator-activated receptors a and b/d (but not g)and liver X receptor activators", 《J ALLERGY CLIN IMMUNOL》 * |
| 孙博强等: "DNCB诱导昆明小鼠特应性皮炎模型的建立", 《生物技术世界》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020156593A1 (en) * | 2019-01-30 | 2020-08-06 | 郑州大学 | Anti-atopic dermatitis protein |
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