阅读说明：本技术 僵蚕水提物及其应用 (Bombyx Batryticatus water extract and application thereof ) 是由 夏庆友 赵萍 孙悦婷 李晓鸿 于 2020-06-08 设计创作，主要内容包括：本发明公开了僵蚕水提物及其应用,通过以水作为提取溶剂提取制得,提取得到的产物具有安全性好,具有较好的自由基清除活性,同时具有酪氨酸酶抑制活性,可以降低LPS刺激后巨噬细胞分泌的TNF-α、IL-6的水平和表达量,同时降低炎性介质COX-2的表达量,具有较好的抗炎的潜力,因此可以作为化妆品原料在抗氧化、美白、抗炎等方面应用,对拓僵蚕的使用价值具有重要意义。(The invention discloses a stiff silkworm aqueous extract and application thereof, wherein the stiff silkworm aqueous extract is prepared by extracting with water as an extraction solvent, and the extracted product has high safety, high free radical scavenging activity and tyrosinase inhibitory activity, can reduce the levels and expression levels of TNF- α and I L-6 secreted by macrophages after L PS stimulation, can reduce the expression level of inflammatory medium COX-2, and has high anti-inflammatory potential, so that the stiff silkworm aqueous extract can be used as a cosmetic raw material to be applied to the aspects of oxidation resistance, whitening, inflammation resistance and the like, and has important significance for developing the use value of stiff silkworms.)

1. Bombyx Batryticatus aqueous extract, characterized by that: the mass fraction of the protein in the water extract is 55-65%, the mass fraction of the sugar is 15-20%, and the mass fraction of the ash is 25-30%.

2. The Bombyx Batryticatus aqueous extract of claim 1, which is prepared by the following method: extracting dried Bombyx Batryticatus powder with water as extraction solvent, collecting extractive solution, concentrating, and drying to obtain silkworm aqueous extract.

3. The aqueous extract of Bombyx Batryticatus of claim 1, wherein: the adding amount of the water is as follows: adding the material to liquid ratio of 30; the stiff silkworm powder is powder with the particle size of less than 50 meshes; the extraction is carried out at 90 ℃ for 1 h.

4. Use of an aqueous extract of Bombyx Batryticatus according to any one of claims 1-3 for preparing an antioxidant material.

5. Use according to claim 4, characterized in that: the antioxidant is DPPH free radical scavenging, ABTS free radical scavenging or hydroxyl free radical scavenging.

6. Use of the aqueous extract of Bombyx Batryticatus of any one of claims 1-4 for preparing tyrosinase monophenolase or tyrosinase diphenolase inhibitor.

7. Use of the Bombyx Batryticatus aqueous extract of any one of claims 1-4 for the preparation of a medicament for reducing the levels of TNF- α and I L-6 secreted by macrophages after L PS stimulation.

8. The use of the Bombyx Batryticatus aqueous extract of any one of claims 1-4 for the preparation of a medicament for reducing the expression level of inflammatory mediators COX-2.

9. Use of the Bombyx Batryticatus aqueous extract of any one of claims 1 to 4 for preparing whitening or anti-inflammatory cosmetic raw materials.

Technical Field

The invention relates to the field of traditional Chinese medicine extraction, in particular to a stiff silkworm aqueous extract and application of the stiff silkworm aqueous extract.

Background

Silkworm (Bombyx mori) is an important economic insect, which is often infected with fungi, bacteria, viruses, etc. during the breeding process. Among fungal diseases, white muscardine caused by beauveria bassiana is common, and the larva after death of beauveria bassiana (balls.) is muscardine silkworm, also called batryticated silkworm. The stiff silkworm serving as a traditional Chinese medicine has a long medicinal history and has the effects of calming endogenous wind, relieving spasm, dispelling wind, relieving pain, reducing phlegm, resolving masses and the like. It contains multiple components, and modern pharmacological studies show that it has a wide range of pharmacological effects. At present, Stiff silkworm is mostly researched on the aspects of the traditional Chinese medicine effects of resisting convulsion, reducing blood sugar and the like, and other aspects are rarely researched.

Therefore, the research on the safety and the efficacy of the stiff silkworm extract is necessary, which is beneficial to developing the use value of the stiff silkworm and provides a thought and theoretical basis for the research and the development and the popularization of an efficacy stiff silkworm product.

Disclosure of Invention

In view of this, the IC of the product obtained by extraction with water as the extraction solvent₅₀295.77 + -127.46 mu g/m L, 0.5% BBE has no eye irritation by chick embryo chorioallantoic membrane test, indicating high safety of BBW.

In order to achieve the above purpose, the invention provides the following technical scheme:

the weight percentage of protein, sugar and ash of the stiff silkworm aqueous extract is 55-65%, 15-20% and 25-30%

Preferably, the preparation method comprises the following steps: extracting dried Bombyx Batryticatus powder with water as extraction solvent, collecting extractive solution, concentrating, and drying to obtain silkworm aqueous extract.

Preferably, the water is added in an amount of 1: adding the material to liquid ratio of 30; the stiff silkworm powder is powder with the particle size of less than 50 meshes; the extraction is carried out at 90 ℃ for 1 h.

The result of DPPH free radical scavenging experiment of the prepared product shows that when the BBW concentration is 0.625mg/m L, DPPH clearance reaches (83 +/-1)%, the concentration is increased, and the clearance is not obviously increased.

Therefore, the invention provides the following technical scheme:

the application of the stiff silkworm aqueous extract in preparing an antioxidant raw material.

Preferably, the oxidation resistance is DPPH radical scavenging, ABTS radical scavenging or hydroxyl radical scavenging.

The whitening effect of BBW is detected through a tyrosinase inhibition experiment. The result shows that BBW has better inhibition effect on tyrosinase monophenolase and tyrosinase diphenolase and has concentration correlation. Semi-inhibitory concentration IC₅₀1.463mg/m L and 3.02mg/m L, respectively.

Therefore, the invention provides the following technical scheme:

the application of the stiff silkworm aqueous extract in preparing tyrosinase monophenolase or tyrosinase diphenolase inhibitor.

E L ISA experiment and real-time fluorescence quantitative PCR experiment show that BBW can significantly reduce the level and expression level of TNF- α and I L-6 secreted by macrophage after L PS stimulation, and has certain concentration correlation.

In order to achieve the purpose, the invention provides the following technical scheme:

the application of the stiff silkworm aqueous extract in preparing the medicine for reducing the levels of TNF- α and I L-6 secreted by macrophages after L PS stimulation.

The application of the stiff silkworm aqueous extract in preparing the medicine for reducing the expression level of inflammatory medium COX-2.

The antibacterial activity of the BBW is detected by using escherichia coli and staphylococcus aureus as strains to be detected and using a nutrient broth dilution method, and the result shows that the BBW extracted by the method in a safe concentration range does not have the capability of inhibiting the growth of the escherichia coli and the staphylococcus aureus. The above results indicate that BBW may have antioxidant, whitening, and anti-inflammatory effects.

In order to achieve the purpose, the invention provides the following technical scheme:

the application of the stiff silkworm aqueous extract in preparing whitening or anti-inflammatory cosmetic raw materials.

The method has the beneficial effects that the batryticated silkworm is extracted by water, the extract contains protein, sugar and ash, the extract has tyrosinase monophenol enzyme or tyrosinase diphenolase inhibiting effect under the combined action of all the components, free radicals are removed, the levels of TNF- α and I L-6 secreted by macrophages after L PS stimulation is reduced, the expression quantity of inflammatory medium COX-2 is reduced, the safety price of the bated silkworm extract is carried out, and the effect screening is carried out on the bated silkworm extract, so that the use value of the bated silkworm is expected to be developed, and thought and theoretical basis are provided for research and popularization of the effect type traditional Chinese medicine cosmetics.

Drawings

In order to make the object, technical scheme and beneficial effect of the invention more clear, the invention provides the following drawings for explanation:

FIG. 1 is a viability curve of BBW versus HaCat cells.

FIG. 2 shows the viability of BBW versus HaCat cells.

FIG. 3 is an observation of the stimulation of the CAM by BBW (A: negative control; B: positive control; C: 0.5% stimulation of the CAM by BBW).

FIG. 4 shows the DPPH & clearance of BBW.

FIG. 5 shows the ABTS-clearance of BBW.

FIG. 6 shows the OH clearance of BBW.

FIG. 7 shows the total antioxidant capacity of BBW (A: antioxidant capacity of BBW at different concentrations; B: standard curve).

FIG. 8 shows the inhibitory activity of BBW on tyrosinase monophenolase activity (A: the content of melanin produced by BBW at different concentrations with increasing reaction time; B: the inhibitory rate of BBW at different concentrations on tyrosinase monophenolase activity).

FIG. 9 shows the inhibitory activity of BBW on tyrosinase-diphenolase activity (A: the content of melanin produced by BBW at different concentrations with increasing reaction time; B: the inhibitory rate of BBW at different concentrations on tyrosinase-diphenolase activity).

FIG. 10 shows the effect of BBW on TNF- α and I L-6 secretion of RAW264.7 macrophage (A: detecting the content of TNF- α by E L ISA method; B: detecting the content of I L-6 by E L ISA method; C: the effect of BBW on the expression level of TNF- α gene; D: the effect of BBW on the expression level of I L-6 gene).

FIG. 11 shows the inhibitory activity of BBW on COX-2 (A: the inhibitory rate of BBW on COX-2; B: the effect of BBW on the expression level of COX-2 gene).

FIG. 12 shows the antibacterial activity of BBW (A: inhibition of E.coli by BBW; B: inhibition of S.aureus by BBW).

Detailed Description

The present invention is further described with reference to the following drawings and specific examples so that those skilled in the art can better understand the present invention and can practice the present invention, but the examples are not intended to limit the present invention.