Application of protein SPINK7 in preparation of medicine for promoting healing of excessive-inflammation wounded area

A wound healing and protein technology, applied in the field of biomedicine, can solve the problems of incompletely clear and unclear mechanisms, and achieve the effect of promoting diabetic wound healing, promoting healing, and reducing inflammatory reactions

Active Publication Date: 2021-02-26
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Studies have found that SPINK7 plays an important role in regulating cell proliferation and apoptosis, migration and invasion, etc., but its mechanism of action is not yet fully understood
[0004] However, the role and mechanism of SPINK7 protein in skin wound healing and chronic refractory wounds represented by diabetic wounds have not been reported so far. Whether SPINK7 can be used as an intervention target to promote excessive inflammation wound healing is still unclear

Method used

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  • Application of protein SPINK7 in preparation of medicine for promoting healing of excessive-inflammation wounded area
  • Application of protein SPINK7 in preparation of medicine for promoting healing of excessive-inflammation wounded area
  • Application of protein SPINK7 in preparation of medicine for promoting healing of excessive-inflammation wounded area

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: SPINK7 inhibits the inflammatory response of keratinocytes in vitro

[0038] 3.1 Experimental method:

[0039] Cells: Human keratinocyte strain HaCaT cells (purchased from ATCC) were cultured in DMEM high glucose medium (Gibco) containing 10% fetal bovine serum.

[0040] SPINK7 lentivirus infection stable clone screening:

[0041] HaCaT cells were plated overnight, and then the cells were infected with lentiviral empty vectors lenti-CMV and lenti-SPINK7 (GenScript Biotechnology Co., Ltd.) at an MOI (multiplicity of infection) of 200. After 24 hours, replace with fresh medium, and continue culturing for 48 hours. Then use puromycin (Beiyuntian Company) to screen the monoclones, and pick the monoclonals after 7 days to expand and culture them for subsequent experiments.

[0042] Western blot detection of SPINK7 expression in HaCaT cells:

[0043] After establishing HaCaT cells stably infected with SPINK7 lentivirus, extract the total protein from them and con...

Embodiment 2

[0051] Example 2: Knockout of SPINK7 leads to excessive inflammatory response and inhibits wound healing in mice

[0052] 2.1 Experimental method:

[0053] Animals: SPINK7 knockout mice and corresponding control wild-type mice, 8-10 in each group. SPINK7 knockout mice were introduced from Saiye Biotechnology Company and bred in the Experimental Animal Center of the Army Medical University of the Chinese People's Liberation Army.

[0054] Establishment of mouse full-thickness skin defect wound model:

[0055] Mice were anesthetized by intraperitoneal injection of 1% pentobarbital sodium, the back was shaved and the skin was prepared with 75% alcohol and povidone iodine, and two circular full-thickness defects with a diameter of 6 mm were prepared on the back skin with a skin biopsy device. Wound.

[0056] Quantitative analysis of wound healing:

[0057] After establishing the wound model, take photos and record the wound at time points such as 0, 1, 3, 5, 7, and 10 days, ca...

Embodiment 3

[0061] Example 3: Local wound injection of SPINK7 eukaryotic expression plasmid inhibits wound inflammation in diabetic mice and promotes diabetic wound healing

[0062] 3.1 Experimental method:

[0063] Plasmid: The eukaryotic expression plasmid of SPINK7 and the control CMV empty plasmid were purchased from Origene Company, amplified and purified according to conventional methods (Molecular Cloning Experiment Guide, October 2013, fourth edition).

[0064] Diabetic mice: C57BL / 6 mice were continuously injected with 50 mg / kg streptozotocin (STZ, Roche) according to body weight for 5 days. After 10 days of stabilization, the blood glucose of the mice was detected, and if the blood glucose value was >20mmol / l, the diabetic mice were successfully modeled.

[0065] Local plasmid injection on the wound surface: Immediately inject 10ug / 100ul of the purified plasmid at the 3 points of the wound edge after the diabetic mouse prepared a circular full-thickness skin defect wound with a...

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Abstract

The invention relates to application of a protein SPINK7 in preparation of a medicine for promoting healing of an excessive-inflammation wounded area. According to the application, an excessive inflammatory reaction of a wounded area of skin can be effectively inhibited, healing of wounded areas, particularly a diabetes wounded area is promoted, a way is provided for promoting healing of chronic inflammatory wounded areas, particularly the diabetes wounded area, and thus, the application has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to the application of SPINK7 protein in the preparation of medicines for promoting the healing of excessive inflammation wounds. Background technique [0002] Inflammation is the body's defense response to injury, which is usually beneficial, but excessive inflammation can lead to impaired skin wound healing. Skin wound healing is a complex and orderly dynamic repair process, which is mainly composed of three distinct and overlapping stages, namely, the inflammatory response stage, the proliferation stage and the remodeling stage. Inflammation is the initiating factor for the smooth healing of skin wounds, but excessive and continuous inflammatory responses will make wounds difficult to heal and form chronic non-healing wounds. The difficulty in healing chronic inflammatory wounds, especially diabetic wounds, is a thorny worldwide medical problem. At present, there ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K48/00A61P17/02A61P29/00A61K8/64A61Q19/00C07K14/81C12N15/12
CPCA61K38/1709A61K48/005A61P17/02A61P29/00A61K8/64A61Q19/00C07K14/8135Y02A50/30
Inventor 王涛粟永萍王军平赵娜龙爽汪国建
Owner ARMY MEDICAL UNIV
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