Method for promoting generation of collagen

Abstract

The invention provides a method for promoting collagen production. The method comprises the step of promoting type III collagen production by using ergothioneine. The application finds that the low-concentration ergothioneine has the effect of promoting the generation of the type III collagen, and particularly, the enhanced effect of promoting the generation of the type III collagen is shown along with the reduction of the concentration of the ergothioneine, so that a new thought is provided for the application of the ergothioneine.

Description

Technical Field

[0001] The present application belongs to the technical field of medicines, cosmetics and medical devices, and specifically, relates to a method for promoting collagen production. Background Art

[0002] Collagen, also known as collagen, is a biological macromolecular protein synthesized by animal cells. It is widely present in the animal body, with abundant content and various types. It has a significant impact on the normal function and damage repair of cells, tissues and even organs.

[0003] The collagen in adult skin tissue is mainly type I and type III collagen. In contrast, the proportion of type III collagen in fetal skin is significantly higher than that in adults. The reduction of type III collagen is associated with skin aging, resulting in a lack of elasticity and regeneration. Supplementation of type III collagen can reduce scar hyperplasia and promote skin regeneration and wound healing. Human type III collagen can effectively promote the transformation of macrophages from type M1 to type M2, the migration ability of L929 cells, and the angiogenesis of human umbilical vein endothelial cells (HUVECs).

[0004] However, the production of type I and type III collagen is not positively correlated. Promoting the production of type I collagen does not necessarily mean that the production of type III collagen can be promoted, and vice versa. Most of the related research focuses on the production of type I collagen, and there are currently few reports on how to promote the production of type III collagen. Summary of the invention

[0005] In view of the problems existing in the prior art, the present application unexpectedly found that low concentrations of ergothioneine have the effect of promoting the production of type III collagen, especially, as the concentration of ergothioneine decreases, it shows an enhanced effect of promoting the production of type III collagen, thereby completing the present invention.

[0006] Specifically, this application involves the following aspects:

[0007] The present application provides a method for promoting the production of type III collagen.

[0008] Further, the method includes using ergothioneine to promote type III collagen production.

[0009] Further, the effective concentration of the ergothioneine is 0.001ng / mL-500ng / mL.

[0010] The present application also provides a method for promoting cells to produce type III collagen.

[0011] Further, the method comprises treating cells with ergothioneine at a concentration of 0.001 ng / mL-500 ng / mL.

[0012] The present application provides the use of ergothioneine in promoting the production of type III collagen.

[0013] Further, the effective concentration of the ergothioneine is 0.001ng / mL-500ng / mL.

[0014] The present application provides a use of ergothioneine as a type III collagen production promoter.

[0015] Further, the effective concentration of ergothioneine in the accelerator is 0.001ng / mL-500ng / mL.

[0016] The present application provides a use of ergothioneine in preparing a type III collagen production promoter.

[0017] Further, the effective concentration of ergothioneine in the accelerator is 0.001ng / mL-500ng / mL.

[0018] Further, the content of ergothioneine in the accelerator is 0.001ng / ml-500ng / ml.

[0019] Furthermore, the administration of the enhancer includes in vivo and / or in vitro.

[0020] Furthermore, the promoter is used to prevent and / or alleviate skin problems associated with decreased type III collagen.

[0021] Furthermore, the promoter includes medicines, medical devices, cosmetics or foods.

[0022] The present application provides a type III collagen production promoter, which comprises ergothioneine in a content of 0.001 ng / mL-500 ng / mL.

[0023] The present application finds that low concentrations of ergothioneine have the effect of promoting the production of type III collagen, and in particular, as the concentration of ergothioneine decreases, an enhanced effect of promoting the production of type III collagen is shown, which provides a new idea for the application of ergothioneine. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a bar graph of the relative expression of type III collagen. DETAILED DESCRIPTION

[0025] The present application is further described below in conjunction with examples. It should be understood that the examples are only used to further describe and illustrate the present application and are not used to limit the present application.

[0026] Unless otherwise defined, the technical and scientific terms in this specification have the same meaning as those generally understood by those skilled in the art. Although similar or identical methods and materials as described herein may be used in experiments or practical applications, materials and methods are described herein below. In the event of a conflict, the present specification including the definitions therein shall prevail. In addition, materials, methods and examples are provided for illustration only and are not restrictive. The present application is further described below in conjunction with specific embodiments, but is not intended to limit the scope of the present application.

[0027] The present application provides a method for promoting the production of type III collagen, which comprises using ergothioneine to promote the production of type III collagen.

[0028] In some embodiments, the effective concentration of the ergothioneine is 0.001ng / ml-500ng / ml, for example, 0.001ng / ml, 0.002ng / ml, 0.003ng / ml, 0.004ng / ml, 0.005ng / ml, 0.006ng / ml, 0.007ng / ml, 0.008ng / ml, 0.009ng / ml, 0.01ng / ml, 0.02ng / ml, 0.03ng / ml, 0.04ng / ml, 0.05ng / ml, 0.06ng / ml, 0.0 7ng / ml, 0.08ng / ml, 0.09ng / ml, 0.2ng / ml, 0.2ng / ml, 0.3ng / ml, 0.4ng / ml, 0.5ng / ml, 0.6ng / ml, 0.7ng / ml, 0.8ng / ml, 0.9ng / ml, 1ng / ml, 2ng / ml, 3ng / ml, 4ng / ml, 5ng / ml, 6ng / ml, 7ng / ml, 8ng / ml, 9ng / ml, 10ng / ml, 15ng / ml, 20ng / ml, 25ng / ml, 30ng / m l, 35ng / ml, 40ng / ml, 45ng / ml, 50ng / ml, 55ng / ml, 60ng / ml, 65ng / ml, 70ng / ml, 75ng / ml, 80ng / ml, 85ng / ml, 90ng / ml, 95ng / m l, 100ng / ml, 150ng / ml, 200ng / ml, 210ng / ml, 220ng / ml, 230ng / ml, 240ng / ml, 250ng / ml, 260ng / ml, 270ng / ml, 280ng / ml, 290 470ng / ml, 480ng / ml, 490ng / ml, 500ng / ml, and any range between these values.

[0029] In the present application, the effective concentration of ergothioneine refers to the effective concentration of ergothioneine that promotes the production of type III collagen, by applying ergothioneine at the effective concentration to a target object, such as a cell or a human body, that needs to produce type III collagen, thereby promoting the production of type III collagen.

[0030] In some embodiments, the administration concentration of the ergothioneine is 0.001ng / ml-500ng / ml. For example, it can be 0.001ng / ml, 0.002ng / ml, 0.003ng / ml, 0.004ng / ml, 0.005ng / ml, 0.006ng / ml, 0.007ng / ml, 0.008ng / ml, 0.009ng / ml, 0.01ng / ml, 0.02ng / ml, 0.03ng / ml, 0.04ng / ml, 0.05ng / ml, 0.06ng / ml, 0.07ng / ml, 0.08ng / ml, 0.09ng / ml, 0.2n g / ml, 0.2ng / ml, 0.3ng / ml, 0.4ng / ml, 0.5ng / ml, 0.6ng / ml, 0.7ng / ml, 0.8ng / ml, 0.9ng / ml, 1ng / ml, 2ng / ml, 3ng / ml, 4 ng / ml, 5ng / ml, 6ng / ml, 7ng / ml, 8ng / ml, 9ng / ml, 10ng / ml, 15ng / ml, 20ng / ml, 25ng / ml, 30ng / ml, 35ng / ml, 40ng / ml, 45 ng / ml, 50ng / ml, 55ng / ml, 60ng / ml, 65ng / ml, 70ng / ml, 75ng / ml, 80ng / ml, 85ng / ml, 90ng / ml, 95ng / ml, 100ng / ml, 150n g / ml, 200ng / ml, 210ng / ml, 220ng / ml, 230ng / ml, 240ng / ml, 250ng / ml, 260ng / ml, 270ng / ml, 280ng / ml, 290ng / ml, 300n 100ng / ml, 100ng / ml, 1020ng / ml, 1040ng / ml, 1060ng / ml, 1080ng / ml, 1090ng / ml, 110ng / ml, 1110ng / ml, 1120ng / ml, 1130ng / ml, 1140ng / ml, 1150ng / ml, 1160ng / ml, 1170ng / ml, 1180ng / ml, 1190ng / ml, 220ng / ml, 2210ng / ml, 2220ng / ml, 2230ng / ml, 2240ng / ml, 225

[0031] In some embodiments, the ergothioneine can be applied to cells or human bodies. The mode of application applied to human bodies includes internal and / or external administration. The external administration can be smearing, external application, etc., and the internal administration can be oral administration, injection, etc.

[0032] The present application also provides a method for promoting cells to produce type III collagen.

[0033] In some embodiments, the method comprises treating cells with ergothioneine at a concentration of 0.001 ng / ml-500 ng / ml, for example, 0.001 ng / ml, 0.002 ng / ml, 0.003 ng / ml, 0.004 ng / ml, 0.005 ng / ml, 0.006 ng / ml, 0.007 ng / ml, 0.008 ng / ml, 0.009 ng / ml, 0.01 ng / ml, 0.02 ng / ml, 0.03 ng / ml, 0.04 ng / ml, 0.05 ng / ml, 0.06 ng / ml ml, 0.07ng / ml, 0.08ng / ml, 0.09ng / ml, 0.2ng / ml, 0.2ng / ml, 0.3ng / ml, 0.4ng / ml, 0.5ng / ml, 0.6ng / ml, 0.7ng / ml, 0.8ng / ml, 0.9ng / ml, 1ng / ml, 2ng / ml, 3ng / ml, 4ng / ml, 5ng / ml, 6ng / ml, 7ng / ml, 8ng / ml, 9ng / ml, 10ng / ml, 15ng / ml, 20ng / ml, 25ng / ml, 30 ng / ml, 35ng / ml, 40ng / ml, 45ng / ml, 50ng / ml, 55ng / ml, 60ng / ml, 65ng / ml, 70ng / ml, 75ng / ml, 80ng / ml, 85ng / ml, 90ng / ml, 95n g / ml, 100ng / ml, 150ng / ml, 200ng / ml, 210ng / ml, 220ng / ml, 230ng / ml, 240ng / ml, 250ng / ml, 260ng / ml, 270ng / ml, 280ng / ml, 2 470ng / ml, 480ng / ml, 490ng / ml, 500ng / ml, and any range between these values.

[0034] The cells include, but are not limited to, fibroblasts, fibrocytes, mesenchymal cells, tenocytes, and epithelial cells.

[0035] The present application also provides a use of ergothioneine to promote the production of type III collagen, wherein the effective concentration of the ergothioneine is 0.001 ng / mL-500 ng / mL.

[0036] The present application also provides a use of ergothioneine as a type III collagen production promoter.

[0037] The present application also provides a use of ergothioneine in preparing a type III collagen production promoter.

[0038] Those skilled in the art will appreciate that the collagen production promoter may use ergothioneine as the sole active ingredient to promote the production of type III collagen, or may contain other active ingredients that jointly promote the production of type III collagen with ergothioneine.

[0039] In some embodiments, the effective concentration of ergothioneine in the collagen production promoter is 0.001ng / ml-10ng / ml, for example, 0.001ng / ml, 0.002ng / ml, 0.003ng / ml, 0.004ng / ml, 0.005ng / ml, 0.006ng / ml, 0.007ng / ml, 0.008ng / ml, 0.009ng / ml, 0.01ng / ml, 0.02ng / ml, 0.03ng / ml, 0.04ng / ml, 0.05ng / ml, 0.06ng / ml, 0.07ng / ml, 0.08ng / ml, 0.09ng / ml, 0.2ng / ml, 0.2ng / ml, 0.3ng / ml, 0.4ng / ml, 0.5ng / ml, 0.6ng / ml, 0.7ng / ml, 0.8ng / ml, 0.9ng / ml, 1ng / ml, 2ng / ml, 3ng / ml, 4ng / ml, 5ng / ml, 6ng / ml, 7ng / ml, 8ng / ml, 9ng / ml, 10ng / ml, 15ng / ml, 20ng / ml, 25ng / ml, 3 0ng / ml, 35ng / ml, 40ng / ml, 45ng / ml, 50ng / ml, 55ng / ml, 60ng / ml, 65ng / ml, 70ng / ml, 75ng / ml, 80ng / ml, 85ng / ml, 90ng / ml, 95 ng / ml, 100ng / ml, 150ng / ml, 200ng / ml, 210ng / ml, 220ng / ml, 230ng / ml, 240ng / ml, 250ng / ml, 260ng / ml, 270ng / ml, 280ng / ml, 2 470ng / ml, 480ng / ml, 490ng / ml, 500ng / ml, and any range between these values.

[0040] In some embodiments, the content of ergothioneine in the collagen production promoter is 0.001ng / ml-500ng / ml. For example, it can be 0.001ng / ml, 0.002ng / ml, 0.003ng / ml, 0.004ng / ml, 0.005ng / ml, 0.006ng / ml, 0.007ng / ml, 0.008ng / ml, 0.009ng / ml, 0.01ng / ml, 0.02ng / ml, 0.03ng / ml, 0.04ng / ml, 0.05ng / ml, 0.06ng / ml, 0.07ng / ml, 0.08ng / ml, 0.09ng / ml, 0.2n g / ml, 0.2ng / ml, 0.3ng / ml, 0.4ng / ml, 0.5ng / ml, 0.6ng / ml, 0.7ng / ml, 0.8ng / ml, 0.9ng / ml, 1ng / ml, 2ng / ml, 3ng / ml, 4 ng / ml, 5ng / ml, 6ng / ml, 7ng / ml, 8ng / ml, 9ng / ml, 10ng / ml, 15ng / ml, 20ng / ml, 25ng / ml, 30ng / ml, 35ng / ml, 40ng / ml, 45 ng / ml, 50ng / ml, 55ng / ml, 60ng / ml, 65ng / ml, 70ng / ml, 75ng / ml, 80ng / ml, 85ng / ml, 90ng / ml, 95ng / ml, 100ng / ml, 150n g / ml, 200ng / ml, 210ng / ml, 220ng / ml, 230ng / ml, 240ng / ml, 250ng / ml, 260ng / ml, 270ng / ml, 280ng / ml, 290ng / ml, 300n 100ng / ml, 100ng / ml, 1020ng / ml, 1040ng / ml, 1060ng / ml, 1080ng / ml, 1090ng / ml, 110ng / ml, 1110ng / ml, 1120ng / ml, 1130ng / ml, 1140ng / ml, 1150ng / ml, 1160ng / ml, 1170ng / ml, 1180ng / ml, 1190ng / ml, 220ng / ml, 2210ng / ml, 2220ng / ml, 2230ng / ml, 2240ng / ml, 225

[0041] Therefore, the present application also provides a type III collagen production promoter, which comprises ergothioneine in an amount of 0.001 ng / mL-500 ng / mL.

[0042] In some embodiments, the collagen production promoter can be applied to cells or to the human body. The administration method for the human body includes in vivo and / or in vitro administration. The in vitro administration can be applied by smearing, external application, etc., and the in vivo administration can be oral administration, injection, etc.

[0043] In some embodiments, the collagen production promoter can be made into cosmetics, medical devices, medicines, foods, etc. For example, the collagen production promoter can be formulated into cosmetics, medical devices, medicines, foods, etc. alone or together with other active ingredients to promote the production of type III collagen.

[0044] In some embodiments, the above-mentioned collagen production promoter can be used to prevent or alleviate skin problems associated with reduced type III collagen. Among them, the skin problems associated with reduced type III collagen can be various types of skin problems caused by reduced type III collagen known in the art, such as reduced skin elasticity, loss of skin elasticity, increased skin wrinkles, sagging skin, slow skin barrier repair, rough skin, etc.

[0045] Example

[0046] The relevant information of the experimental materials and experimental equipment used in the examples is shown in Table 1.

[0047] Table 1

[0048] Experimental materials and equipment source Primary human dermal fibroblasts Lifeline DMEM medium, fetal bovine serum, 96-well cell culture plate Thermo CCK reagent Tongren Bio Human Collagen Type III ELISA Kit BIO-SWAMP Cell culture incubator Thermofisheri160 Inverted microscope Leica DMI1 Biological Safety Cabinets Labconco Class II ELISA reader BioTek 800TS

[0049] Example

[0050] 1. Sample Preparation

[0051] The ergothioneine solid powder was prepared into a solution with the basic culture solution, and the concentration was adjusted to 200 ng / mL, 2 ng / mL, 0.2 ng / mL, and 0.02 ng / mL.

[0052] 2. Test of Type III Collagen Production in Primary Human Dermal Fibroblasts

[0053] (1)Planning

[0054] Human primary dermal fibroblasts were cultured in 5% CO 2 The cells were expanded and cultured in fibroblast medium at 37°C. When the fibroblasts cultured in vitro grew to 80-90% confluence, they were digested with trypsin and seeded into 96-well plates. The cells were cultured in the 96-well plates for 48 hours.

[0055] (2) Cell grouping

[0056] The cell of step (1) is grouped according to thioneine concentration, 200ng / mL, 2ng / mL, 0.2ng / mL, 0.02ng / mL, respectively, and a blank control group and a positive control group are separately established, wherein the blank control group is serum-free culture fluid, and the positive control group is 10ng / mL TGF-β1. Every group of 3 holes is repeated. Corresponding culture fluid is added to the cell wells of each group, and culture is continued for 48h in an incubator.

[0057] (3) Detection

[0058] The cell culture supernatant was taken and the type III collagen content was determined by referring to the Human Collagen III ELISA kit. The type III collagen content of the sample group and the positive control group was statistically compared with the blank control group. All results were expressed as Mean ± SD, and P < 0.05 was considered to be statistically significant. The statistical results are shown in Table 2 and Figure 1 shown.

[0059] 3. Experimental results

[0060] From Table 2 and Figure 1 It can be seen from the data that the positive control has a significant promoting effect on the synthesis of type III collagen in fibroblasts. Ergothioneine has a significant promoting effect on the production of type III collagen at a concentration of 200ng / mL-0.02ng / mL, and at a concentration below 200ng / mL, its promoting effect has a certain upward trend. Among them, the promoting effect on the production of type III collagen is most significant at a concentration of 2ng / mL.

[0061] Table 2

[0062]

Claims

1. A method for promoting the production of type III collagen, the method comprising using ergothioneine to promote the production of type III collagen, wherein the effective concentration of the ergothioneine is 0.001 ng / mL-500 ng / mL.

2. A method for promoting cells to produce type III collagen, the method comprising treating cells with ergothioneine at a concentration of 0.001 ng / mL-500 ng / mL.

3. The purpose of ergothioneine in promoting the production of type III collagen, wherein the effective concentration of the ergothioneine is 0.001 ng / mL-500 ng / mL.

4. The invention discloses a method for using ergothioneine as a promoter for the production of type III collagen, wherein the effective concentration of ergothioneine in the promoter is 0.001 ng / mL to 500 ng / mL.

5. The invention relates to a method for preparing a type III collagen generation promoter, wherein the effective concentration of ergothioneine in the promoter is 0.001 ng / mL to 500 ng / mL.

6. according to the purposes described in claim 4 or 5, in described accelerator, the content of ergothioneine is 0.001ng / mL-500ng / mL.

7. The use according to claim 4 or 5, wherein the administration of the enhancer includes in vivo and / or in vitro.

8. The use according to claim 4 or 5, wherein the promoter is used to prevent and / or alleviate skin problems associated with decreased type III collagen.

9. The use according to claim 4 or 5, wherein the promoter comprises medicine, medical device, cosmetic or food.

10. A type III collagen production promoter, comprising ergothioneine in an amount of 0.001 ng / mL to 500 ng / mL.

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