Application of human hepatocyte growth factor gene in eczema treatment and microneedle medical instrument

A technology of hepatocyte growth factor and gene, which is applied in the application of human hepatocyte growth factor gene in the treatment of eczema and in the field of microneedle medicine and equipment, can solve the problems of eczema prone to repeated attacks and serious side effects, and achieve anti-inflammatory, itching, and inhibition of skin inflammation. The effect of itching and good application prospects

Active Publication Date: 2021-10-22
ACADEMY OF MILITARY MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because eczema is prone to recurrent attacks, patients need to re

Method used

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  • Application of human hepatocyte growth factor gene in eczema treatment and microneedle medical instrument
  • Application of human hepatocyte growth factor gene in eczema treatment and microneedle medical instrument
  • Application of human hepatocyte growth factor gene in eczema treatment and microneedle medical instrument

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Obtaining of recombinant plasmid vector carrying human hepatocyte growth factor gene

[0055] 1. Cell preparation

[0056] Using the mammalian cell expression vector pVAX1 (Invitrogen, catalog number V260-20, full length 2999bp) as the backbone, the plasmid pHGF carrying the human hepatocyte growth factor gene was constructed by itself.

[0057] The vector pVAX1 contains CMV promoter, BGH polyA, kanamycin resistance gene and pUC ori, among them, there is a multiple cloning site between the CMV promoter and BGH polyA, including T7 promoter and sequentially connected restriction enzyme sites point.

[0058] The CMV promoter is located at 137-724bp; the T7 promoter is located at 664-683bp; the multiple cloning site: 696-811bp;

[0059] BGH poly A: 829-1053bp; kanamycin resistance gene: 1226-2020bp; pUC ori: 2320-2993bp.

[0060] Human hepatocyte growth factor gene (full length 2.3Kb) is placed between restriction site BamH I and XbaI, thereby obtaining recombinant plasm...

Embodiment 2

[0068] Embodiment 2: Preparation of pHGF microneedle patch

[0069] The microtemplate method is one of the most commonly used preparation methods for microneedle casting. The general procedure of this method is to first rely on microelectromechanical system (MEMS) technology to make a microneedle positive mold with specific parameters, then use polydimethylsiloxane (PDMS) to copy the microneedle positive mold to obtain a microneedle template, and finally The required solution is dropped into the microneedle template, and the air in the micropores and the solution is removed by vacuuming or heating, so that the solution can better fill the pinhole groove and ensure its mechanical strength. Soluble microneedles.

[0070] Formula screening: Through research, it is found that when the concentration of hyaluronic acid increases from low to high, the hardness of the microneedles gradually increases. When the concentration reaches 250mg / mL, the hardness is good, and the hardness dec...

Embodiment 3

[0073] Embodiment 3: The therapeutic effect of human hepatocyte growth factor gene on skin eczema of BABL / c mice

[0074] Animal model: male BABL / c mice, after adaptive feeding for 3 days, were randomly divided into 3 groups: eczema model group, pHGF microneedle group and pHGF subcutaneous injection group. Remove rat hair 2cm×2cm. On d1, d2, and d3, apply 0.5% DNCB acetone solution to the skin of the shaved area on the back of the mouse for sensitization; then apply 1% DNCB acetone solution to the skin of the shaved area on the back of the mouse twice a week for stimulation, and stop after 28 days.

[0075] Administration method: Microneedle administration (only PBS solution) in the shaved area of ​​mice in the model group, once a week; pHGF microneedles of 0.1 mg (dissolved in 100 μL of PBS) in the shaved area of ​​mice in the pHGF microneedle group Administration, 1 time a week; 0.1 mg of pHGF subcutaneously injected into the shaved area of ​​the mice in the pHGF subcutaneous...

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Abstract

The invention provides application of a human hepatocyte growth factor gene in eczema treatment and a microneedle medical instrument. The human hepatocyte growth factor gene is constructed into an eukaryotic expression vector, a virus vector and a gene expression cassette or minicircle DNA (Deoxyribose Nucleic Acid), and can obviously play a role in resisting eczema and related symptoms after being locally introduced subcutaneously by adopting a microneedle or injection mode and the like. Therefore, the human hepatocyte growth factor gene has an extremely good application prospect in the aspect of eczema treatment, and especially in the form of a microneedle instrument, not only can the eczema symptom be inhibited, but also the side effect is small.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to the application of a human hepatocyte growth factor gene in the treatment of eczema and the microneedle medical device. Background technique [0002] Eczema is a skin inflammatory reaction caused by various internal and external factors, often accompanied by severe itching. The vicious cycle of "itching-scratching" further aggravates eczema, making it difficult to heal and easy to relapse. At present, the medicines for treating eczema are mainly aimed at its pruritus symptoms, adopting a multi-drug combination mode, such as local application of anxiolytic drug doxepin and antiallergic drug glucocorticoid ointment. In addition, there are also traditional Chinese medicines for clearing away heat and dampness, nourishing blood and moistening dryness, and acupuncture to treat eczema, and have achieved certain therapeutic effects. However, because eczema is prone to recurrent attacks, pati...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K38/18A61K9/00A61K47/36A61M37/00A61P17/00A61P17/04A61P29/00
CPCA61K48/005A61K48/0008A61K38/1833A61P17/00A61P29/00A61P17/04A61K9/0021A61K47/36A61M37/0015A61M2037/0053A61M2037/0061
Inventor 张庆林杜丽胡申辉卢育新程晓晨
Owner ACADEMY OF MILITARY MEDICAL SCI
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