Application of periplaneta americana growth factor PDGF to preparation of damage repairing medicines

A technology of Periplaneta americana and growth factors, applied in the direction of growth factors/inducing factors, platelet-derived growth factors, applications, etc., to achieve the effect of promoting the healing of mechanical injuries

Pending Publication Date: 2019-12-20
SICHUAN GOODDOCTOR PANXI PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant information about the growth factor PDGF of the American cockroach. Therefore, the relevant information of the PDGF gene is obtained through the transcriptome and expressed in the prokaryotic. After purification, the PDGF of the American cockroach is obtained.

Method used

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  • Application of periplaneta americana growth factor PDGF to preparation of damage repairing medicines
  • Application of periplaneta americana growth factor PDGF to preparation of damage repairing medicines
  • Application of periplaneta americana growth factor PDGF to preparation of damage repairing medicines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Prokaryotic expression of Periplaneta americana growth factor PDGF

[0025] 1 Experimental materials

[0026] Strains: Escherichia coli DH5α, Escherichia coli BL21

[0027] Vector: pET32a(+)

[0028] Restriction enzymes: BamH I, Not I

[0029] Culture medium: LB medium

[0030] 2 Experimental methods

[0031] (1) Screen out Periplaneta americana PDGF from transcriptome data;

[0032] (2) carry out ORF, signal peptide, codon analysis to Periplaneta americana PDGF gene;

[0033] (3) Use ORF Finder online software to find the largest ORF in the PDGF gene of Periplaneta americana; and use SignalP4.1Serverj to analyze the signal peptide of the ORF, and remove the signal peptide; send the gene that removes the signal peptide to the company for codon optimization, and synthesized;

[0034] (4) The vector pET32a(+) was double-digested with restriction endonucleases BamH I and Not I. The double-digestion system is shown in Table 1, and incubated at 37° C. for 15...

Embodiment 2

[0044] Example 2 Identification and Purification of Periplaneta Americana Growth Factor PDGF Expression Form

[0045] 1 Experimental materials and methods

[0046] 1.1 Experimental materials

[0047] Strain: Escherichia coli BL21 transformed with recombinant vector pET32a(+)-PDGF

[0048] Culture medium: LB medium

[0049] 1.2 Experimental method

[0050] (1) Inoculate 500 microliters of Escherichia coli BL21 transformed with the recombinant vector pET32a(+)-PDGF into 50 mL of LB medium, culture on a shaker at 37°C for 4 hours, then add IPTG to a final concentration of 0.5mM, and culture at 25°C 6h, to induce expression.

[0051](2) Centrifuge 50mL of the induced bacterial solution at 8000rpm for 10min, remove the supernatant, collect the precipitate, resuspend the precipitate with 5mL of 50mM pH7.4PB buffer, and crush the cells in a cell grinder, 3s plus / 7s stop, 45 %P, 30 min. The crushed cells were centrifuged at 8000rpm for 10min to obtain the supernatant and the pre...

Embodiment 3

[0064] Example 3 Periplaneta americana growth factor PDGF repairs mechanical damage in mice

[0065] 1 Materials and methods

[0066] 1.1 Experimental materials

[0067] Drugs: refolded PDGF pure product, Kangfuxin solution (Sichuan Good Doctor Panxi Pharmaceutical Co., Ltd.), recombinant bovine basic fibroblast growth factor solution for external use (recombinant bovine FGF) (Beifuji, Zhuhai Yisheng Biopharma Limited);

[0068] Experimental animals: SPF grade Yunnan mice, male, weighing 30-35 g, 8 weeks old, purchased from Chengdu Dashuo Experimental Animal Co., Ltd., certificate number SCXK (Sichuan) 2015-030.

[0069] 1.2 Experimental method

[0070] (1) Establishment of mechanical injury model in mice: firstly, depilate the back of the mouse with 8% sodium sulfide, establish the wound model after depilation, and cut out 1cm on the back with scissors 2 Left and right wounds, mechanical damage model was established.

[0071] (2) Experimental grouping: the mice were divi...

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Abstract

The invention discloses an application of a periplaneta americana growth factor PDGF to preparation of damage repairing medicines. The periplaneta americana growth factor PDGF is obtained through thesteps of leading in escherichia coli for prokaryotic expression, and then performing identification and purification on expressed products. The purified PDGF is further used for a mechanical damage model mouse for repairing activity determination, and the result shows that the periplaneta americana PDGF has high repairing activity for mechanical damage of a mouse. The repairing activity of the periplaneta americana growth factor PDGF is researched, and a new thinking is provided for development of a novel damage repairing medicine.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of a growth factor PDGF of the American cockroach in the preparation of damage repairing medicine. Background technique [0002] With the continuous development of modern molecular biology and cell biology techniques, people's understanding of wound repair has gradually deepened. The process of tissue wound repair is not only the process of various cell proliferation, differentiation, migration, apoptosis and disappearance, but also the result of a network interaction between a series of different types of cells, structural proteins, growth factors and protein kinases. It has been confirmed that growth factors play a significant role in burn wound repair and regulate various stages of wound repair. Growth factors not only promote the transformation of fibroblasts into myofibroblasts, but also initiate apoptosis signals as the wound heals to promote the re...

Claims

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Application Information

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IPC IPC(8): C07K14/49C12N15/12C12N15/70A61K38/18A61P17/02
CPCC07K14/49C12N15/70A61P17/02A61K38/00Y02A50/30
Inventor 刘越飞鲍锦库岳碧松
Owner SICHUAN GOODDOCTOR PANXI PHARMA
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