Compositions with telomere shortening inhibition efficacy, telomere shortening inhibition products comprising the same, and their applications.

Abstract

This application relates to the field of biochemical technology, and particularly to compositions with telomere shortening inhibition effects, telomere shortening inhibition products containing the same, and their applications. The compositions include one or more of the following: Gastrodia elata powder, Codonopsis pilosula powder, and mangosteen extract; and Panax ginsenoside A. This composition can effectively delay telomere shortening and effectively protect telomeres, thereby achieving anti-aging effects.

Description

Technical Field

[0001] This application relates to the field of biochemical technology, and in particular to compositions having telomere shortening inhibition efficacy, telomere shortening inhibition products containing the same, and their applications. Background Technology

[0002] Telomeres are specialized structures at the ends of chromosomes that play a crucial role in maintaining chromosome stability and preventing DNA damage. With cell division, telomeres gradually shorten, leading to cell aging and death. Skin, as the largest organ in the human body, acts as a barrier, protecting the body from the loss of water and nutrients. However, with age, telomeres in skin cells gradually shorten, leading to skin aging and the appearance of sagging skin and wrinkles.

[0003] Several studies have reported on drug screening and combination therapies targeting telomere shortening. For example, some studies have found that using telomerase inhibitors can slow telomere shortening, thereby delaying cellular aging. In addition, some studies have explored the use of antioxidants, DNA repair enzymes, and other drugs to protect telomeres.

[0004] Given that finding substances that can protect telomeres is of great significance for delaying cell aging and preventing related diseases, this application is made. Summary of the Invention

[0005] Based on this, one or more embodiments of this application provide a composition with telomere shortening inhibition effect, a telomere shortening inhibition product containing the same, and its application. The technical solutions include the following:

[0006] One or more embodiments of this application provide a composition having telomere shortening inhibition effect, the composition comprising:

[0007] One or more of the following: Gastrodia elata powder, Codonopsis pilosula powder, and mangosteen extract; and,

[0008] Panlongshenol A.

[0009] In some embodiments of this application, the composition, when comprising 0.5 μmol-3 μmol of Pantothecin A, includes one or more of the following (1) to (3):

[0010] (1) 450μg-550μg of Gastrodia elata powder;

[0011] (2) 450μg-550μg of Codonopsis pilosula powder;

[0012] (3) 4μg-13μg mangosteen extract.

[0013] In some embodiments of this application, the composition, when comprising 0.7 μmol-1.2 μmol of Pantothecin A, includes one or more of the following (1) to (3):

[0014] (1) 485μg-515μg of Gastrodia elata powder;

[0015] (2) 485μg-515μg of Codonopsis pilosula powder;

[0016] (3) 8μg-12μg mangosteen extract.

[0017] One or more embodiments of this application provide a composition with telomere shortening inhibition effect or the application of Panlongshenol A in the preparation of telomere shortening inhibition products.

[0018] In some embodiments of this application, the telomere shortening inhibitor is used to reduce the expression level of p16 and / or the telomere shortening inhibitor is used to reduce the expression level of p21.

[0019] In some embodiments of this application, the telomere shortening inhibitory product is used orally and / or topically.

[0020] One or more embodiments of this application provide a telomere shortening inhibition product, the telomere shortening inhibition product comprising the composition having telomere shortening inhibition efficacy.

[0021] In some embodiments of this application, the telomere shortening inhibitory product comprises 0.5 μM-3 μM of Pantothecin A and one or more of the following (I) to (III):

[0022] (I) 450μg / mL-550μg / mL Gastrodia elata powder;

[0023] (II) 450μg / mL-550μg / mL Codonopsis pilosula powder;

[0024] (III) 4μg / mL-13μg / mL mangosteen extract.

[0025] In some embodiments of this application, the telomere shortening inhibitor product comprises 0.7 μM-1.2 μM of Pantothecin A and one or more of the following (I) to (III):

[0026] (I) 485μg / mL-515μg / mL Gastrodia elata powder;

[0027] (II) 485μg / mL-515μg / mL Codonopsis pilosula powder;

[0028] (III) 8μg / mL-12μg / mL mangosteen extract.

[0029] One or more embodiments of this application provide a method for inhibiting telomere shortening in cells, the method comprising the step of contacting target cells with the composition having telomere shortening inhibitory efficacy or the telomere shortening inhibitory product.

[0030] Compared with traditional technologies, this application has the following advantages:

[0031] This application provides a specific composition, namely, a composition of Panlongshen A with one or more of Gastrodia elata powder, Codonopsis pilosula powder and mangosteen extract, which can effectively delay telomere shortening and effectively protect telomeres, thereby achieving anti-aging. Attached Figure Description

[0032] To more clearly illustrate the technical solutions in the embodiments of this application and to more completely understand this application and its beneficial effects, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0033] Figure 1 The results of each experiment show the detection results of the cytotoxic effect of HaCaT cells.

[0034] Figure 2a and Figure 2b The figure shows the effects of each experimental group on the expression of the senescence genes CDKN2A (encoding p16 protein) and CDKN1A (encoding p21 protein) in HaCaT cells. Detailed Implementation

[0035] The present application will be further described in detail below with reference to the accompanying drawings, embodiments, and examples. It should be understood that these embodiments and examples are for illustrative purposes only and are not intended to limit the scope of the present application. The purpose of providing these embodiments and examples is to enable a more thorough and comprehensive understanding of the disclosure of the present application. It should also be understood that the present application can be implemented in many different forms and is not limited to the embodiments and examples described herein. Those skilled in the art can make various modifications or alterations without departing from the spirit of the present application, and the equivalent forms obtained also fall within the protection scope of the present application. Furthermore, numerous specific details are set forth in the following description to provide a fuller understanding of the present application. It should be understood that the present application can be implemented without one or more of these details.

[0036] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the specification of this application is for descriptive purposes only and is not intended to be limiting of the application.

[0037] Unless otherwise stated or in case of contradiction, the terms or phrases used herein shall have the following meanings:

[0038] The terms "and / or," "or / and," and "and / or" as used herein include any one of two or more of the related listed items, as well as any and all combinations of the related listed items. These arbitrary and all combinations include any two related listed items, any more related listed items, or a combination of all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or," "or / and," and "and / or," it should be understood that in this application, the technical solution undoubtedly includes technical solutions connected by "logical AND," and also undoubtedly includes technical solutions connected by "logical OR." For example, "A and / or B" includes three parallel solutions: A, B, and A+B. For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, a technical solution that is connected by "logical OR"), as well as any and all combinations of A, B, C, and D, that is, combinations of any two or three of A, B, C, and D, and also combinations of all four of A, B, C, and D (that is, a technical solution that is connected by "logical AND").

[0039] In this application, the terms "multiple", "various", "multiple times", "multi-dimensional", etc., unless otherwise specified, refer to a quantity greater than or equal to 2. For example, "one or more" means one or more than or equal to two.

[0040] The terms “combinations of,” “any combination of,” and “any combination of” used in this article include all suitable combinations of any two or more of the listed items.

[0041] In this document, the term "suitable" as used in phrases such as "suitable combination," "suitable method," and "any suitable method" refers to the ability to implement the technical solution of this application, solve the technical problem of this application, and achieve the expected technical effect of this application.

[0042] In this document, terms such as “preferred,” “better,” “more suitable,” and “ideal” are merely used to describe implementation methods or examples that achieve better results, and should be understood not to limit the scope of protection of this application.

[0043] In this application, terms such as "further," "even further," and "particularly" are used to describe purposes and indicate differences in content, but should not be construed as limiting the scope of protection of this application.

[0044] In this application, "optionally," "optionally," and "optional" mean that something is optional, that is, it means that it is selected from either "with" or "without." If there are multiple "optional" entries in a technical solution, unless otherwise specified, and there are no contradictions or mutual constraints, each "optional" entry shall be independent.

[0045] In this application, the terms "first aspect," "second aspect," "third aspect," "fourth aspect," etc., are used for descriptive purposes only and should not be construed as indicating or implying relative importance or quantity, nor should they be construed as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first," "second," "third," "fourth," etc., serve only as a non-exhaustive enumeration and should be understood not to constitute a closed limitation on quantity.

[0046] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0047] In this application, numerical intervals (i.e., numerical ranges) are involved. Unless otherwise specified, the selected numerical distributions within the aforementioned numerical intervals are considered continuous and include the two endpoints (i.e., the minimum and maximum values) of the numerical range, as well as every value between these two endpoints. Unless otherwise specified, when a numerical interval refers only to integers within that interval, it includes the two endpoint integers of the numerical range, as well as every integer between the two endpoints. In this document, this is equivalent to directly listing every integer. For example, if t is an integer selected from 1 to 10, it means that t is any integer selected from the group of integers consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, and 10. Furthermore, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0048] Unless otherwise specified, the temperature parameters in this application are permitted to be either constant-temperature treatment or variations within a certain temperature range. It should be understood that the constant-temperature treatment allows temperature fluctuations within the precision range of the instrument control, such as ±5℃, ±4℃, ±3℃, ±2℃, or ±1℃.

[0049] In this application, %(w / w) and wt% both represent weight percentage, %(v / v) refers to volume percentage, and %(w / v) refers to mass-volume percentage.

[0050] All references to this application are incorporated herein by reference as if each document were individually incorporated herein by reference. Unless they conflict with the purpose and / or technical solution of this application, all cited references are incorporated herein by reference in their entirety and for all purposes. When references are cited in this application, the definitions of relevant technical features, terms, nouns, phrases, etc., are also incorporated herein by reference. Examples and preferred embodiments of the cited technical features may also be incorporated herein by reference, but only to the extent that they enable the implementation of this application. It should be understood that when the cited content conflicts with the description in this application, this application shall prevail or modifications shall be made adaptably to the description in this application.

[0051] A first aspect of this application provides a composition having telomere shortening inhibition effect, the composition comprising:

[0052] One or more of the following: Gastrodia elata powder, Codonopsis pilosula powder, and mangosteen extract; and,

[0053] Panlongshenol A.

[0054] The composition of this application may be a composition of Gastrodia elata powder and Panlongshenol A, a composition of Codonopsis pilosula powder and Panlongshenol A, a composition of mangosteen extract and Panlongshenol A, or a composition of Gastrodia elata powder, Codonopsis pilosula powder and Panlongshenol A, a composition of Codonopsis pilosula powder, mangosteen extract and Panlongshenol A, or a composition of Gastrodia elata powder, Codonopsis pilosula powder, mangosteen extract and Panlongshenol A.

[0055] In some examples of this application, the composition includes one or more of the following (1) to (3) when comprising 0.5 μmol-3 μmol of Pantothecin A:

[0056] (1) 450μg-550μg of Gastrodia elata powder;

[0057] (2) 450μg-550μg of Codonopsis pilosula powder;

[0058] (3) 4μg-13μg mangosteen extract.

[0059] In the composition provided in this application that satisfies the above-mentioned proportions of each component, if the amount of one component changes, the amounts of the other components will change proportionally. For example, if the composition includes 0.5 mol-3 mol of Pantothecin A, it may include 450 g-550 g of Gastrodia elata powder, 450 g-550 g of Codonopsis pilosula powder, and / or 4 g-13 g of Mangosteen extract.

[0060] In some examples of this application, the composition of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panlongshenol A, comprises 450 μg-550 μg (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg) of Gastrodia elata powder, i.e., the ratio of Panlongshenol A to Gastrodia elata powder is 0.5 μmol-3 μmol: 450 μg-550 μg.

[0061] In some examples of this application, the composition of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panax notoginseng A, comprises 450 μg-550 μg (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg) of Codonopsis pilosula powder, i.e., the ratio of Panax notoginseng A to Codonopsis pilosula powder is 0.5 μmol-3 μmol: 450 μg-550 μg.

[0062] In some examples of this application, the composition of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panax ginsenoside A, comprises 4 μg-13 μg (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg) of mangosteen extract, i.e., the ratio of Panax ginsenoside A to mangosteen extract is 0.5 μmol-3 μmol: 4 μg-13 μg.

[0063] In some examples of this application, the composition of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panax notoginseng A, comprises 450 μg-550 μg (e.g., 45 μg) of Panax notoginseng A. The dosage ratio of Panlongshen A, Gastrodia elata powder, and Codonopsis pilosula powder is 0.5μmol-3μmol:450μg-550μg:450μg-550μg.

[0064] In some examples of this application, the composition of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panax notoginseng A, comprises 450 μg- 550 μg (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg) of Codonopsis pilosula powder and 4 μg-13 μg (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg) of mangosteen extract, i.e., the ratio of Panlongshen A, Codonopsis pilosula powder and mangosteen extract is 0.5 μmol-3 μmol: 450 μg-550 μg: 4 μg-13 μg.

[0065] In some examples of this application, the composition of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panax notoginseng A, comprises 450 μg- 550 μg (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg) of Gastrodia elata powder and 4 μg-13 μg (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg) of mangosteen extract, i.e., the ratio of Panlongshen A, Gastrodia elata powder and mangosteen extract is 0.5 μmol-3 μmol: 450 μg-550 μg: 4 μg-13 μg.

[0066] In some examples of this application, the compositions of this application, when comprising 0.5 μmol-3 μmol (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μmol) of Panax notoginseng A, comprise 450 μg-550 μg (e.g., 450, 460, 470, 480, 490, 500, 510, 520 μg) of Panax notoginseng A. The dosage of Panlongshen A, Gastrodia elata powder, Codonopsis pilosula powder and mangosteen extract is 0.5μmol-3μmol: 450μg-550μg: 450μg-550μg: 4μg-13μg.

[0067] In some examples of this application, the composition includes one or more of the following (1) to (3) when comprising 0.7 μmol-1.2 μmol of Pantothecin A:

[0068] (1) 485μg-515μg of Gastrodia elata powder;

[0069] (2) 485μg-515μg of Codonopsis pilosula powder;

[0070] (3) 8μg-12μg mangosteen extract.

[0071] A second aspect of this application provides a composition having telomere shortening inhibition effect, or the application of Panlongshenol A in the preparation of telomere shortening inhibition products.

[0072] In some examples of this application, the telomere shortening inhibitory product is used to reduce the expression level of p16.

[0073] In some examples of this application, the telomere shortening inhibitory product is used to reduce the expression level of p21.

[0074] In some embodiments of this application, the telomere shortening inhibitory product is used orally and / or topically.

[0075] A third aspect of this application provides a telomere shortening inhibition product, the telomere shortening inhibition product comprising the composition having telomere shortening inhibition efficacy.

[0076] This application does not specifically limit the types of products that inhibit telomere shortening; they can be food, pharmaceuticals, or skincare products.

[0077] In some examples of this application, the telomere shortening inhibitory product comprises 0.5 μM-3 μM of Pantothecin A and one or more of the following (I) to (III):

[0078] (I) 450μg / mL-550μg / mL Gastrodia elata powder;

[0079] (II) 450μg / mL-550μg / mL Codonopsis pilosula powder;

[0080] (III) 4μg / mL-13μg / mL mangosteen extract.

[0081] In some examples of this application, the telomere shortening inhibitory product of this application includes 450 μg / mL - 550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL) of Gastrodia elata powder and 0.5 μM - 3 μM (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM) of Panax ginseng A.

[0082] In some examples of this application, the telomere shortening inhibitory product of this application includes 450 μg / mL - 550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL) of Codonopsis pilosula powder and 0.5 μM - 3 μM (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM) of Panax ginsenoside A.

[0083] In some examples of this application, the telomere shortening inhibitory product of this application includes 4 μg / mL-13 μg / mL (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg / mL) of mangosteen extract and 0.5 μM-3 μM (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM) of ginsenoside A.

[0084] In some examples of this application, the telomere shortening inhibitory products of this application include Gastrodia elata powder at concentrations of 450 μg / mL to 550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL), and Gastrodia elata powder at concentrations of 450 μg / mL to 550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL). Codonopsis pilosula powder (0 μg / mL) and Panlongshen phenol A (0.5 μM-3 μM, e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM).

[0085] In some examples of this application, the telomere shortening inhibitory products of this application include 450 μg / mL - 550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL) Codonopsis pilosula powder, 4 μg / mL -13 μg / mL (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg / mL) mangosteen extract and 0.5 μM-3 μM (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM) pangolin A.

[0086] In some examples of this application, the telomere shortening inhibitory product of this application comprises 450 μg / mL -550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL) Gastrodia elata powder, 4 μg / mL-13 μg / mL (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg / mL) mangosteen extract and 0.5 μM-3 μM (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM) Pantothecin A.

[0087] In some examples of this application, the telomere shortening inhibitory products of this application include 450 μg / mL-550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL) of Gastrodia elata powder, 450 μg / mL-550 μg / mL (e.g., 450, 460, 470, 480, 490, 500, 510, 520, 530, 540, 550 μg / mL) of Codonopsis pilosula powder, and 4 μg / mL... -13 μg / mL (e.g., 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 μg / mL) mangosteen extract and 0.5 μM-3 μM (e.g., 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3 μM) pangolin A.

[0088] In some examples of this application, the telomere shortening inhibitory product comprises 0.7 μM-1.2 μM of Pantothecin A and one or more of the following (I) to (III):

[0089] (I) 485μg / mL-515μg / mL Gastrodia elata powder;

[0090] (II) 485μg / mL-515μg / mL Codonopsis pilosula powder;

[0091] (III) 8μg / mL-12μg / mL mangosteen extract.

[0092] A fourth aspect of this application provides a method for inhibiting telomere shortening in cells, the method comprising the step of contacting a target cell with the composition having telomere shortening inhibitory effect or the telomere shortening inhibitory product.

[0093] The embodiments of this application will be described in detail below with reference to examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of this application. For experimental methods in the following embodiments where specific conditions are not specified, please refer to the guidelines given in this application, or follow experimental manuals or conventional conditions in the art, or follow the conditions recommended by the manufacturer, or refer to experimental methods known in the art.

[0094] In the specific embodiments described below, the measurement parameters involving raw material components may have slight deviations within the weighing accuracy range unless otherwise specified. Temperature and time parameters are subject to acceptable deviations due to instrument testing accuracy or operational precision.

[0095] Gastrodia elata powder (purchased from Guangdong Qingyunshan Pharmaceutical Co., Ltd.) is a traditional Chinese medicine. It has been found that Gastrodia elata has effects such as improving cognition and memory, improving sleep, regulating blood lipids, blood sugar and blood pressure, and improving liver damage. It contains a large amount of gastrodin, a natural medicinal component that has a significant effect on the human nervous system, clearing nerve disorders and improving nerve function. Furthermore, gastrodin can regulate oxidative stress and inflammatory responses by activating the AMPK / Nrf2 pathway, improving lipid metabolism in non-alcoholic fatty liver disease. Codonopsis pilosula powder (purchased from Guangdong Qingyunshan Pharmaceutical Co., Ltd.) contains active ingredients such as polysaccharides, alkyne glycosides, and total saponins. Codonopsis pilosula polysaccharides have effects such as improving immunity, fatigue resistance, anti-stress, anti-inflammation, antioxidant properties, and improving gastrointestinal function. Codonopsis pilosula alkyne glycosides can effectively protect the gastric mucosa from damage. Total saponins of Codonopsis pilosula can exert anti-inflammatory and immune-boosting effects. Mangosteen extract (purchased from Shinyaku Co., Ltd., Japan) is rich in active ingredients such as flavonoids, xanthones, anthocyanins, and proanthocyanidins, which give it anti-inflammatory, analgesic, antioxidant, anticancer, and antibacterial physiological activities, and thus it is widely used in food, medicine, chemical industry and many other fields.

[0096] Spiranthol A, purchased from WuXi AppTec, has the molecular formula C0. 20 H 22 O3, with a relative molecular mass (MW) of 310.39, has the following structural formula:

[0097] .

[0098] Example 1: Dilution of single substances and composite substances

[0099] The test substances were dissolved in ultrapure water to prepare original stock solutions with a concentration of 100 mg / mL for Gastrodia elata powder, 50 mg / mL for Codonopsis pilosula powder, 100 mg / mL for mangosteen extract, and 100 mM for Panlongshen A.

[0100] During the efficacy verification process of Examples 2 to 4, the working concentrations of the original reservoir design were used for each intervention group as shown in Table 1 below.

[0101] Table 1. Analytical Substances and Their Working Concentrations Used in Each Experimental Group

[0102]

[0103] Example 2: Detection of the cytotoxicity of single substances and combined substances against HaCaT cells

[0104] Experimental steps:

[0105] (1) Cell passage culture: Human keratinocytes (HaCaT) were used in the experiment. The cells adhered to the culture flask and grew to cover 80% of the bottom area of ​​the culture flask. When passaged, the original culture medium was poured out, washed twice with PBS, digested with 0.25% trypsin for 2 min, the trypsin was discarded, and the cells were allowed to stand. The cells were digested until they shrank and became rounded and gradually detached from the culture flask. Fresh DMEM complete medium (90% DMEM + 10% FBS) with 3 times the amount of trypsin was added, and the cells were collected by pipetting and centrifugation at 1100 rpm for 5 min. The supernatant was discarded, and the cells were resuspended in fresh culture medium. Finally, the cells were resuspended in 1 mL of culture medium and counted for expansion culture.

[0106] (2) Cell plating: When HaCaT cells reach the logarithmic growth phase, they are digested and counted at a density of 1.1 × 10⁻⁶. 4 Inoculate 100 μL / mL of each sample into a 96-well plate (with the edge wells filled with sterile PBS), and place the plate in a constant temperature incubator at 37°C, 5% CO2, and saturated humidity.

[0107] (3) Cell intervention: Take the original stock solution of the test substance in Example 1 and prepare the working solution using DMEM complete medium. When the cells in the 96-well plate reach 80% confluence, add 100 μL of DMEM complete medium containing the corresponding working concentration of the test substance in Table 1 to each well of the experimental group, and add 100 μL of DMEM complete medium to each well of the blank group. Set up 3 replicate wells for each group. After the intervention, place the 96-well plate in a constant temperature incubator at 37°C, 5% CO2 and saturated humidity.

[0108] (4) Cell viability assay (ATP): After culturing cells for 24 h, discard the original culture medium in the 96-well plate, do not wash the cells, add 110 μL of ATP working solution directly, and incubate at 37 °C for 30 min. Observe the cells under a microscope to see if they become round, and repeatedly pipette 10-15 times to fully lyse the cells. Then, transfer 70 μL of the test liquid to a new white opaque microplate and measure the bioluminescence value of each well using a microplate reader.

[0109] (5) Cell viability calculation: Cell viability (%) = luminescence value of sample well ÷ luminescence value of blank well × 100%.

[0110] (6) Test results: such as Figure 1 Compared with the blank control group, the groups treated with the test substance showed no toxicity to HaCaT cells.

[0111] Example 3: Effects of single substances and combined substances on telomere length in a TBHP-induced HaCaT cell injury model

[0112] Experimental steps:

[0113] (1) Cell passage culture: Refer to step (1) in Example 2 above.

[0114] (2) Group setup: Cells in good growth condition were seeded at 700,000 per dish in 6cm cell culture dishes containing fresh DMEM complete medium. A blank control group, a model group, a positive control group, and an experimental group were set up. Referring to Table 1, each experimental group was pre-protected with the corresponding working concentration of the test substance for 24 hours. For example, experimental group 1 was pre-protected with the original stock solution of Gastrodia elata powder and Panlongshen A, resulting in working concentrations of 500 μg / mL and 1 μM, respectively. The positive control group was pre-protected with 6 μM (working concentration) of cycloastragalool (an effective telomerase activator that can prolong telomeres) for 24 hours.

[0115] The model group, positive control group, and experimental group were treated with tert-butanol hydrogen peroxide (TBHP) at a final concentration of 0.15 mM for 6 h, and samples were collected afterward. The blank control group was not treated with tert-butanol hydrogen peroxide.

[0116] (3) Genomic DNA extraction: A genomic DNA extraction kit (purchased from Tiangen Biotech, catalog number DP304) was used, following the kit instructions. The steps are as follows:

[0117] Prepare a cell suspension, centrifuge at 10,000 rpm for 1 min, discard the supernatant, add 200 μL of buffer GA to the resulting precipitate, and vortex until completely resuspended. Add 20 μL of proteinase K solution to the suspension, mix well, then add 200 μL of buffer GB, mix thoroughly by inverting, and incubate at 70°C for 10 min until the solution becomes clear. Add 200 μL of anhydrous ethanol, shake thoroughly for 15 s, transfer the resulting solution to adsorption column CB3, centrifuge at 12000 rpm for 30 s, discard the waste liquid, add 500 μL of buffer GD, centrifuge at 12000 rpm for 30 s, discard the waste liquid, add 600 μL of buffer PW, centrifuge at 12000 rpm for 30 s, discard the waste liquid, repeat the PW addition step again, discard the waste liquid, centrifuge at the same speed for 2 min, place the adsorption column at room temperature for several minutes, then transfer to a clean centrifuge tube, add 70 μL of ddH2O dropwise, place at room temperature for 3 min, centrifuge at 12000 rpm for 2 min, collect the solution in the centrifuge tube, and then determine the concentration.

[0118] (4) Relative telomere length qPCR: Dilute the genomic DNA to 6 ng / μL and select Tel and RPLP0 (internal control) as primers for the reaction system.

[0119] The total volume of the reaction system was 20 μL, including: 7.4 μL H2O, 10 μL Supermix, 0.8 μL each of 10 μmol / L upstream and downstream primers, and 1 μL DNA.

[0120] The reaction program was set as follows: pre-denaturation at 95℃ for 15 min, followed by denaturation at 94℃ for 15 s, and annealing extension at 54℃ for 2 min, for 40 cycles.

[0121] Each qPCR reaction was performed in triplicate, with three replicates. Primer sequences are shown in Table 2.

[0122] Table 2. Telomere gene primer sequences

[0123]

[0124] (4) The relative telomere length is calculated as follows: ΔCt = Ct(telomere) - Ct(internal parameter), relative telomere length = 2 -△Ct .

[0125] (5) Experimental results: Compared with the blank control group (relative telomere length of 1.00), the telomere length of the model group was significantly reduced, with a relative telomere length of 0.70, indicating that the TBHP-induced telomere shortening model of HaCaT cells was successfully constructed. The relative telomere lengths of each experimental group are shown in Table 3. Compared with the model group, each experimental group showed better protective effects, and the combined substance was significantly better than the single substance.

[0126] Table 3

[0127]

[0128] Example 4: Effects of single substances and combined substances on aging markers in the TBHP-induced HaCaT telomere shortening model

[0129] Experimental steps:

[0130] (1) Cell passage culture, same as step (1) in Example 3.

[0131] (2) The group setting is the same as step (2) in Example 3.

[0132] (3) RNA extraction: Discard the cell culture medium, wash the cells with 1×PBS, and add 500 μL of RNA-easy (Vazyme, Nanjing, China) to each well of a 6-well plate to fully cover the cell surface. Then, pipette the cells and collect them into 1.5 mL RNase-free centrifuge tubes. Add 200 μL of ddH2O to each tube, vortex, and incubate at room temperature for 15 min. Centrifuge at 11200 rpm for 15 min at room temperature. Slowly aspirate 500 μL of supernatant and add an equal volume of isopropanol. Vortex, incubate at room temperature for 10 min, and centrifuge at 11200 rpm for 10 min at room temperature. Discard the supernatant and resuspend the precipitate with 75% ethanol. Vortex for 30 s and centrifuge at 9100 rpm for 3 min at room temperature. After drying at room temperature, add an appropriate amount of RNase-free ddH2O to dissolve the precipitate. Use a NanoDrop2000 (ThermoFisher) to determine the RNA concentration and quality.

[0133] (4) Reverse transcription and qPCR: Using a reverse transcription kit (TIANGEN, Beijing, China), 1 μg of total RNA was synthesized into cDNA according to the manufacturer's instructions. The synthesis system consisted of 4 μL of 5×FastKing-RTSuperMIX, 8 μL of RNA, and RNase-free ddH2O to a final volume of 20 μL. The reaction program was 42℃ for 15 min, followed by 95℃ for 3 min. Specific qPCR primers were designed using β-actin as an internal reference gene. The reaction system consisted of 10 μL of 2×TaqPro Universal SYBR qPCRMasterMix (Vazyme, Nanjing, China), 0.4 μL each of 10 μM forward and reverse primers, 1 μL of cDNA, and ddH2O to a final volume of 20 μL. The reaction program was 95℃ for 30 s pre-denaturation, 95℃ for 10 s denaturation, 60℃ for annealing and extension for 30 s, for 40 cycles, followed by melting curve analysis. The primers used for the gene were synthesized by Beijing Tianyi Huiyuan Biotechnology Co., Ltd., and the primer sequences are shown in Table 4.

[0134] Table 4. Primer sequences for aging genes

[0135]

[0136] (5) Test results: such as Figure 2a , Figure 2b Compared with the blank control group, the expression of CDKN2A and CDKN1A genes in the model group was significantly increased, indicating that TBHP-induced telomere shortening in HaCaT cells ultimately leads to cellular senescence. Compared with the model group, astragalool, Codonopsis pilosula powder, mangosteen extract, and Panax ginseng A all reduced the expression of CDKN2A and CDKN1A genes; Gastrodia elata powder significantly reduced the expression of CDKN2A gene, but had no significant effect on CDKN1A gene expression. The combined substance significantly reduced the expression of CDKN2A and CDKN1A genes, and was significantly superior to each individual substance.

[0137] Conclusion: The combination drug composed of Gastrodia elata powder, Codonopsis pilosula powder, mangosteen extract and Panlongshen A can prolong the telomere length of human skin keratinocytes, thereby inhibiting cell aging.

[0138] The technical features of the above-described embodiments and examples can be combined in any suitable manner. For the sake of brevity, not all possible combinations of the technical features in the above-described embodiments and examples are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0139] The embodiments described above are merely illustrative of several implementation methods of this application, intended to facilitate a detailed understanding of the technical solutions of this application, but should not be construed as limiting the scope of protection of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the scope of protection of this application. Furthermore, it should be understood that after reading the above teachings of this application, those skilled in the art can make various alterations or modifications to this application, and the equivalent forms obtained also fall within the scope of protection of this application. It should also be understood that technical solutions obtained by those skilled in the art based on the technical solutions provided in this application through logical analysis, reasoning, or limited experimentation are all within the scope of protection of the appended claims. Therefore, the scope of protection of this patent application should be determined by the content of the appended claims, and the specification and drawings can be used to interpret the content of the claims.

Claims

1. A composition having anti-aging effects, characterized in that, The composition comprises 0.5 × 10 -3 μmol-3×10 -3 In the case of μmol of Pantothenic A, one or more of the following (1) to (3) are included: (1) 450μg-550μg of Gastrodia elata powder; (2) 450μg-550μg of Codonopsis pilosula powder; (3) 4μg-13μg mangosteen extract; The gastrodia elata powder was purchased from Guangdong Qingyunshan Pharmaceutical Co., Ltd. The Codonopsis pilosula powder was purchased from Guangdong Qingyunshan Pharmaceutical Co., Ltd. The mangosteen extract was purchased from Nippon Shinyaku Co., Ltd.

2. The composition with anti-aging effects according to claim 1, characterized in that, The composition comprises 0.7 μmol × 10 -3 -1.2×10 -3 In the case of μmol of Pantothenic A, one or more of the following (1) to (3) are included: (1) 485μg-515μg of Gastrodia elata powder; (2) 485μg-515μg of Codonopsis pilosula powder; (3) 8μg-12μg mangosteen extract.

3. The composition with anti-aging effects according to any one of claims 1 to 2, or the use of Panlongshen A in the preparation of anti-aging products.

4. The application according to claim 3, characterized in that, The anti-aging product is used to reduce the expression level of p16; and / or the anti-aging product is used to reduce the expression level of p21.

5. The application according to any one of claims 3 to 4, characterized in that, The anti-aging products can be used orally and / or topically.

6. An anti-aging product, characterized in that, The anti-aging product includes the composition with anti-aging effects as described in claim 1.

7. The anti-aging product according to claim 6, characterized in that, The anti-aging product includes 0.5μM-3μM of Pantothecin A and one or more of the following (I) to (III): (I) 450μg / mL-550μg / mL Gastrodia elata powder; (II) 450μg / mL-550μg / mL Codonopsis pilosula powder; (III) 4μg / mL-13μg / mL mangosteen extract.

8. The anti-aging product according to any one of claims 6 to 7, characterized in that, The anti-aging product comprises 0.7μM-1.2μM of Pantothecin A and one or more of the following (I) to (III): (I) 485μg / mL-515μg / mL Gastrodia elata powder; (II) 485μg / mL-515μg / mL Codonopsis pilosula powder; (III) 8μg / mL-12μg / mL mangosteen extract.

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