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Preparation method and application of male silk moth exosomes

A technology of male silkworm moth and exosomes, which is applied in the field of exosomes to achieve the best market prospects and clinical value

Active Publication Date: 2020-09-11
HUBEI WEILAIJIAYUAN HIGH TECH AGRI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These kits do not require special equipment. With the continuous upgrading of products, the extraction efficiency and purification effect are gradually improved. However, there is currently a lack of kits for insect exosome extraction.

Method used

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  • Preparation method and application of male silk moth exosomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Extraction of male silkworm moth exosomes and preparation of freeze-dried powder

[0023] Take healthy living male silkworm moths, soak them in normal saline, break them with a tissue homogenizer to make homogenate, and carry out the following steps:

[0024] (1) Centrifuge the homogenate at 3000g for 30min, collect the supernatant, and use the precipitated part for other purposes;

[0025] (2) Centrifuge the 10000g supernatant of step (1) for 100 minutes, collect the supernatant, and use the precipitated part for other purposes;

[0026] (3) Centrifuge the 100000 g supernatant of step (2) for 100 minutes, discard the supernatant, and collect the precipitate;

[0027] (4) After adding the precipitate of step (3) to PBS for remixing, centrifuge at 120,000 g for 120 min, discard the supernatant, and collect the precipitate; the precipitate is suspended in PBS;

[0028] (5) Add mannitol to the PBS suspension of step (4) to a final concentration of 10%, and after mixing, pas...

Embodiment 2

[0031] Example 2: Preparation of a medicine containing male silkworm moth exosomes in the form of an aqueous solution to treat injuries

[0032] The main components (w / v) of the water preparation are: 1-5% glycerin, 1-10% propylene glycol, thickening agent; 1-2% Tween, 0.1% ethyl p-hydroxybenzoate, and 60ml PBS buffer Dissolve, then add 10-50μg of male silkworm moth exosomes freeze-dried powder, add 5ml 0.01M phosphate buffer, mix and make up PBS to a final volume of 100ml, filter with 0.45μm filter to sterilize and insolubles, complete the water preparation form The preparation process of the product containing male silkworm moth exosomes.

[0033] The prepared water containing male silkworm moth exosomes can be used as a spray, lotion, etc., and can be sprayed or applied.

Embodiment 3

[0034] Example 3: Preparation of a product in the form of an emulsion containing male silkworm moth exosomes

[0035] The emulsified form of the product can be prepared by different methods of oil-in-water or water-in-oil, such as 10% glycerol, 2% Tween 80, 2% cholesterol, 5% lecithin, 0.2% ethyl paraben and 70ml PBS buffer Mix and heat to homogenize and emulsify. After cooling to 37-45℃, add 10-50μg of male silkworm moth exosomes freeze-dried powder, add 5ml 0.01M phosphate buffer, mix and add water to a final volume of 100ml, homogenize and filter by 0.45μm Membrane filtration eliminates bacteria and insolubles, and completes the preparation process of a product containing male silkworm moth exosomes in the form of an emulsion preparation.

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Abstract

The invention discloses a preparation method and application of a male silkworm moth exosome, which belong to the field of a biotechnology. The method comprises the steps of preparing a healthy male silkworm moth living body into a homogenate, carrying out differential centrifugation, and purifying with super speed at low temperature to obtain the exosome with high purity. An animal skin injury experiment curative effect is favorable, and the exosome can be prepared into freeze-dried powder so as to be stored for a long time, and is matched with different ingredients to form different preparation products so as to have an excellent market prospect and clinic value.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to the extraction and application of biologically active exosomes from healthy living male silkworm moths. Background technique [0002] As early as the Tang and Song Dynasties, the male silkworm moth was used by the emperors as a precious common tonic for invigorating the kidney, anti-aging, and prolonging life. Li Shizhen, a well-known medical scientist in the Ming Dynasty, called the male silkworm moth a national treasure of divine insects in the Compendium of Materia Medica. Insects have obvious effects on invigorating the liver and kidney, impoting yang, anti-aging, preventing Alzheimer's disease, preventing male prostate hyperplasia, and regulating female endocrine disorders. There are a dazzling array of male silkworm moth kidney and aphrodisiac products on the market, but no products for treating wounds, ulcers and scalds. The reason may be that the effective ingredients f...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K35/64A61P17/00A61P17/02A61K8/98A61Q19/00A61Q19/02
CPCA61K8/987A61K35/63A61K2800/84A61Q19/00A61Q19/02
Inventor 尹维坊敖明章赵新红
Owner HUBEI WEILAIJIAYUAN HIGH TECH AGRI CO LTD
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