Composition for improving mitochondrial function and application thereof

Abstract

The invention belongs to the technical field of cosmetics, and provides a composition for improving a mitochondrial function and application thereof. The composition comprises prinsepia utilis royle total flavone extract and decapeptide-4 in a mass ratio of (100: 1)-(125: 1). The composition can well recover mitochondrial membrane potential and improve the mitochondrial function, so that the cell activity is improved, and the action effect is better than that of any raw material in the composition which is independently used. Mitochondrial membrane potential is a key factor participating in mitochondria to generate ATP, and when mitochondrial functions are impaired, the mitochondrial membrane potential is reduced, which is also an obvious characteristic in the early stage of cell apoptosis. The composition disclosed by the invention can play a role in improving mitochondrial function and cell activity by recovering mitochondrial membrane potential, relieving mitochondrial damage and maintaining cell mitochondrial homeostasis, and has great application potential in the large health fields of biotechnology, cosmetics, adjuvant therapy of skin diseases and the like.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics, and particularly relates to a composition for improving mitochondrial function and an application thereof. Background Art

[0002] Life activities are inseparable from the fine regulation and energy supply of mitochondria. Mitochondria are not only responsible for producing most of the energy required by cells, but also participate in regulating cell growth, differentiation and death. The normal functioning of mitochondria is essential for maintaining the normal physiological functions of cells and is closely related to the physiological functions of cells and a variety of skin diseases.

[0003] In the skin, mitochondria provide energy for skin metabolism, participate in the regulation of skin physiological processes, and are closely related to maintaining skin homeostasis, proliferation and differentiation, hair growth, and signal transduction. Mitochondria provide energy for the proliferation and differentiation of skin cells, and their downstream transcription factors participate in regulating the proliferation and differentiation of cells. Abnormal mitochondrial function can lead to imbalance of skin homeostasis, weakened resistance, and even delayed wound healing. Mitochondria are also involved in the occurrence and development of many skin diseases, such as systemic lupus erythematosus, psoriasis, atopic dermatitis, and Cockein's syndrome. During the growth process, hair is also extremely dependent on mitochondria for rapid energy supply through aerobic respiration. When mitochondrial function is defective, hair regeneration will encounter obstacles. In a mitochondrial DNA damage experiment, mice with mitochondrial dysfunction showed symptoms of increased skin wrinkles and hair loss.

[0004] When mitochondrial dysfunction occurs, it can seriously affect the health and physiological functions of the skin. Therefore, maintaining healthy mitochondrial function is extremely important for skin homeostasis and health. At present, the main compounds used to improve mitochondrial function are chemically synthesized compounds, such as fisetin, vitamin C, glutathione and other substances. These compounds mainly improve mitochondrial function by scavenging mitochondrial oxygen free radicals, or by activating autophagy to clear dysfunctional mitochondria and retain mitochondria with normal functions, rather than improving mitochondrial function, and they mainly focus on the impact of mitochondrial function on diseases such as diabetes and lung injury.

[0005] Prinia prickly ash, also known as pyracantha, is a shrub of the genus Prickly ash in the Rosaceae family, originating from Yunnan, China. It is often used to clear away heat and detoxify, promote blood circulation and reduce swelling, and treat snake bites, so prinia prickly ash is also known as the "auspicious tree" and the "king of flowers". Prinia prickly ash is rich in active ingredients such as flavonoids, polysaccharides and high-quality oils. Studies have shown that flavonoids in prinia prickly ash have anti-inflammatory and soothing effects.

[0006] Decapeptide-4 can act on high mobility group protein HMGB1, inhibit inflammatory response, and prevent or treat atopic dermatitis. However, in the prior art, there is no public report that the combination of total flavonoids extract of Prinus utilis and decapeptide-4 can improve mitochondrial function, and its specific concentration ratio has not been publicly reported. Summary of the invention

[0007] In view of the above technical problems, the present invention mixes the total flavonoids extract of Prinus utilis with decapeptide-4 to prepare a composition. The composition can stabilize the mitochondrial membrane potential and provide a solution for improving the mitochondrial function of the skin. The composition can be added to skin care and beauty products to improve skin problems, and has great application potential in assisting the treatment of skin diseases.

[0008] In order to achieve the above object, the technical solution adopted by the present invention is as follows:

[0009] In a first aspect, the present invention provides a composition for improving mitochondrial function, comprising a total flavonoids extract of Prinus utilis and decapeptide-4 in a mass ratio of 100:1 to 125:1.

[0010] Preferably, the Principia principe total flavonoids extract is the Principia principe seed meal extract after the Principia principe oil is removed.

[0011] Preferably, the mass percentage of total flavonoids in the Prinus utilis total flavonoids extract is 30% to 60%.

[0012] More preferably, the mass percentage of total flavonoids in the total flavonoids extract of Prinia prinia is 56%. In the present invention, only the total flavonoids extract of Prinia prinia with a mass percentage of 56% is used as an example for illustration, and the total flavonoids extract of Prinia prinia with other contents not exemplified can achieve the same technical effect.

[0013] Preferably, the purity of decapeptide-4 is 95%.

[0014] In a second aspect, the present invention provides use of the above composition for improving mitochondrial function in cosmetics.

[0015] Preferably, the cosmetic is any one of lotion, emulsion, essence, essence water, face cream or face mask.

[0016] More preferably, the lotion is a spray or an aqueous solution;

[0017] More preferably, the essence water is a spray or an aqueous solution.

[0018] More preferably, the mask is in gel form.

[0019] Mitochondrial membrane potential is a key factor in mitochondrial ATP production. When mitochondrial function is impaired, it will manifest as a decrease in mitochondrial membrane potential, which is also an obvious feature of early apoptosis. Compared with the prior art, the present invention has at least the following beneficial effects:

[0020] 1. The composition can be well used in beauty and skin care products by stabilizing the mitochondrial membrane potential of skin fibroblasts, improving skin mitochondrial function, and reducing skin problems caused by cell mitochondrial damage.

[0021] 2. The composition is simple, consisting of only two raw materials, with few preparation steps, which can effectively improve industrialization efficiency and reduce energy consumption.

[0022] 3. The composition is natural, non-irritating and highly safe to use.

[0023] 4. The composition is more environmentally friendly and scientific than chemically synthesized substances. The flavonoids of Prinia prittari are derived from the seed meal of Prinia prittari seeds after oil extraction, which improves the utilization rate of natural plants and reduces the loss of effective substances in natural plants. Decapeptide-4 can effectively reduce skin inflammatory reactions and prevent or treat atopic dermatitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 This is a diagram showing the relative change in mitochondrial membrane potential in Test Example 1 of the present invention. DETAILED DESCRIPTION

[0025] The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. However, the following examples are only simplified examples of the present invention and do not represent or limit the scope of protection of the present invention. The scope of protection of the present invention shall be subject to the claims.

[0026] Unless otherwise specified, the raw materials and materials used in the examples of the present invention are purchased through general commercial channels.

[0027] The sources of the raw materials, materials and instruments involved in the following examples or comparative examples are as follows:

[0028] The total flavonoids extract of Principia utilis was prepared according to the method in Example 4 of Chinese invention patent CN108553527B, and the mass percentage of total flavonoids in the extract was 56%;

[0029] Decapeptide-4 comes from Hemei Biotechnology (Zhejiang) Co., Ltd. The purity of decapeptide-4 in this product is 95%. It is a cyclic peptide composed of ten amino acids with the sequence of CIPRFYLFYC.

[0030] Mitochondrial membrane potential detection kit (JC-1), from Shanghai Bio-Tech Biotechnology Co., Ltd.;

[0031] Hydrogen peroxide (H2O2) was from Sinopharm Chemical Reagent Co., Ltd.;

[0032] Flow cytometer, from Becton, Dickinson and Company (BD);

[0033] Human primary fibroblasts (HDFa) and complete fibroblast culture medium were obtained from the American Type Culture Collection (ATCC);

[0034] CO2 incubator, biosafety cabinet, and centrifuge were all from Thermo Fisher Scientific, USA;

[0035] Inverted microscope, from Olympus Corporation, Japan;

[0036] Fully automatic cell counter, from Nexcelom, USA.

[0037] <Examples 1 to 2>

[0038] According to the formula in Table 1, the total flavonoids extract of Prinus utilis and decapeptide-4 were stirred and mixed uniformly at room temperature to prepare a composition for improving mitochondrial function.

[0039] Table 1. Mass ratio of each composition in Examples 1 to 2

[0040] Total flavonoids extract from Prinus utilis Decapeptide-4 Example 1 100 0.8 Example 2 100 1

[0041] <Comparative Examples 1 to 6>

[0042] According to the preparation method in Examples 1-2, the total flavonoids extract of Prinus prinus and decapeptide-4 were mixed to prepare a mixture of the two, or only the total flavonoids extract of Prinus prinus or decapeptide-4 was weighed. The proportions of each comparative example are shown in Table 2.

[0043] Table 2. Mass ratio of each composition in Comparative Examples 1 to 6

[0044] Total flavonoids extract from Prinus utilis Decapeptide-4 Comparative Example 1 100 0 Comparative Example 2 0 0.5 Comparative Example 3 0 0.8 Comparative Example 4 0 1 Comparative Example 5 150 0.5 Comparative Example 6 100 1.5

[0045] <Application Example 1 and Application Comparative Examples 1 to 5>

[0046] In order to better verify the application of the compositions obtained in the examples and comparative examples in cosmetics, Example 2, Comparative Example 1, Comparative Example 4, Comparative Example 5, and Comparative Example 6 were selected to prepare the formula in the actual essence, and the specific formula is shown in Table 3. Example 2 corresponds to Application Example 1, and Comparative Example 1, Comparative Example 4, Comparative Example 5, Comparative Example 6 and the blank group correspond to Comparative Examples 1 to 5 in sequence.

[0047] Table 3. Actual formula composition of essence (unit: g)

[0048]

[0049] According to the formula in Table 3, the specific preparation process is as follows:

[0050] Step 1, add the raw materials of phase A into the main preparation tank, stir and mix evenly, and heat to 75-85°C;

[0051] Step 2, add the phase B raw material to the pre-mixing tank, heat to 75℃-85℃, stir and mix evenly, then add to the main mixing tank, maintain the temperature at 75-85℃, and mix evenly. Start cooling;

[0052] Step 3, when the temperature drops to 45°C or below, add the raw materials of phase C, mix well, and discharge the material to obtain the product.

[0053] <Test Example 1>

[0054] There is a close relationship between mitochondrial membrane potential (MMP) and mitochondrial dysfunction. Mitochondrial membrane potential is the potential difference caused by the different ion concentrations on both sides of the mitochondrial inner membrane. It reflects the integrity of mitochondrial function and is a sensitive indicator for evaluating mitochondrial function. JC-1 is a cationic lipid fluorescent dye that exists in two forms, monomers and polymers. It is an ideal fluorescent probe widely used to detect mitochondrial membrane potential ΔΨm. When the membrane potential is normal, JC-1 enters the mitochondrial matrix through the polarity of the mitochondrial membrane to form a polymer that emits red fluorescence. When the membrane potential decreases, JC-1 is released from the membrane to form a monomer that reflects green fluorescence. Therefore, the relative ratio of red and green fluorescence is often used to measure changes in mitochondrial membrane potential.

[0055] This test example uses human primary fibroblasts HDFa as a model, and the compositions in Examples 1 to 2 and Comparative Examples 1 to 6 are used to determine the restorative effect on the mitochondrial membrane potential of human primary fibroblasts induced by hydrogen peroxide, in order to evaluate their improvement effect on the mitochondrial function of cells after modeling. The main reagents used include: fibroblast complete culture medium, trypsin, hydrogen peroxide H2O2, phosphate buffer PBS, mitochondrial membrane potential detection kit (JC-1); the main instruments used include: CO2 incubator, inverted microscope, biological safety cabinet, fully automatic cell counter, flow cytometer. The specific experimental process is as follows:

[0056] First, HDFa human primary fibroblasts with good cell morphology and in logarithmic growth phase were prepared with complete culture medium at 4×10 4 / mL concentration of cell suspension; then 2mL per well was inoculated in a 6-well plate, and cultured in an incubator at 37°C, 5% CO2 for 24h, the supernatant was discarded, and then a blank group, a model group and a sample group were set up respectively. 2mL of H2O2 solution with a concentration of 600μmol / L was added to each well of the model group and the sample group, and 2mL of complete culture medium was added to the blank group, and the cells were incubated in an incubator at 37°C, 5% CO2 for 2h, the supernatant was discarded, and the cells were washed twice with PBS solution. 2mL of fresh complete culture medium was added to the blank group and the model group, and 2mL of the test substance prepared with complete culture medium was added to the sample group, respectively, and cultured in an incubator at 37°C, 5% CO2 for 24h. Among them, the test substances are the compositions in Examples 1 to 2 and Comparative Examples 1 to 6, and the concentrations are shown in Table 4 below. After culturing for 24 hours, the supernatant was discarded, 500 μL of trypsin was added to each well and placed in an incubator for 3 minutes, 1 mL of complete medium was added to each well to terminate digestion, and the cells were collected by centrifugation. The mitochondrial membrane potential distribution of the collected cells was detected by flow cytometry using a JC-1 kit. The operating steps were strictly performed in accordance with the instructions of the kit to obtain the mitochondrial membrane potential ΔΨm of each composition in Examples 1 to 2 and Comparative Examples 1 to 6. The mitochondrial membrane potential ΔΨm is the ΔΨm relative to the red / green fluorescence ratio. Figure 1 . Figure 1 In the table, * indicates that the difference between the sample group (example and comparative example) and the model group (H2O2) or the sample group is P<0.05, that is, the difference is significant; ** indicates that the difference between the sample group and the model group or the sample group is P<0.01, that is, the difference is very significant; *** indicates that the difference between the sample group and the model group or the sample group is P<0.001, that is, the difference is extremely significant; ### indicates that the difference between the model group and the control group (Ctrl) is P<0.001, that is, the difference is extremely significant.

[0057] Table 4. Concentration of each test substance

[0058] Total flavonoids extract from Prinus utilis Decapeptide-4 Example 1 0.10％ 8ppm Example 2 0.10％ 10ppm Comparative Example 1 0.10％ 0 Comparative Example 2 0 5ppm Comparative Example 3 0 8ppm Comparative Example 4 0 10ppm Comparative Example 5 0.15％ 5ppm Comparative Example 6 0.10％ 15ppm

[0059] from Figure 1 It can be seen that the compositions of Examples 1 to 2 can significantly improve the decrease in mitochondrial membrane potential of fibroblasts after hydrogen peroxide induction, and can restore the membrane potential to a certain extent, and the effect is better than the compositions in Comparative Examples 1 to 6. That is, when the total flavonoids extract of Prinus utilis or decapeptide-4 is used alone or in an excessive ratio, the effect is poor, and the compositions of Examples 1 to 2 have significant differences compared with the total flavonoids extract of Prinus utilis alone (Comparative Example 1). Therefore, when the mass ratio of the total flavonoids extract of Prinus utilis to decapeptide-4 is 100:1 to 125:1, it has a better stabilizing effect on the mitochondrial membrane potential.

[0060] <Test Example 2>

[0061] Test method: Under normal circumstances, adult subjects use the test product continuously for 28 days according to the instructions to evaluate whether the test product has moisturizing and anti-wrinkle effects and whether it is suitable for sensitive skin.

[0062] Subjects: A total of 37 valid subjects completed the assessment, including healthy Chinese men and women with sensitive skin (lactic acid stinging screening), including 6 males and 31 females, aged 37 to 60 years old, with an average age of 51.22±5.90 years old, who met the voluntary inclusion and exclusion criteria.

[0063] Area assessed: Face.

[0064] How to use: Apply an appropriate amount of this product evenly on the face, avoiding the skin around the eyes, and massage gently in circles in the same direction until absorbed. Frequency of use: once in the morning and once in the evening.

[0065] Use period: 28 days.

[0066] Evaluation period: before using the product (D0), 14 days after using the product (D14), and 28 days after using the product (D28).

[0067] Evaluation method: Visioscan @ VC20plus is used for facial image acquisition and skin texture analysis, skin moisture content measurement instrument Corneometer @ CM825 is used to detect the moisture content of the skin's stratum corneum, and the subjects self-assess whether it is suitable for sensitive skin.

[0068] (1) Increase in moisture content of the stratum corneum (cheeks)

[0069] During the test period, the increase in the moisture content of the stratum corneum of the skin (cheek) of the application examples and the comparative examples is shown in Table 5 below:

[0070] Table 5. Increase in moisture content of the stratum corneum of each application example (cheek) (unit: %)

[0071]

[0072]

[0073] From the above results, it can be seen that there is a significant difference in the increase in the moisture content of the stratum corneum (cheek) of the skin in Application Example 1 compared with Application Comparative Examples 1 to 4, indicating that the composition for improving mitochondrial function in Example 2 can indirectly improve the skin condition, thereby having a good moisturizing effect. However, there is still a significant difference between Application Comparative Examples 1 to 4 and Application Comparative Example 5, which may be because the improvement effect of mitochondrial membrane potential is only one of the factors affecting the moisture content of the stratum corneum (cheek), and the total flavonoids extract of Prinus utilis and decapeptide-4 themselves also have good skin moisturizing effects.

[0074] (2) Reduction in skin wrinkle volume (cheeks)

[0075] During the test period, the skin wrinkle volume reduction (cheek) of each application example is shown in Table 6 below:

[0076] Table 6. Skin wrinkle volume reduction in each application example (cheek) (unit: %)

[0077] Detection time Application Example 1 Application Comparative Example 1 Application Comparative Example 2 Application Comparative Example 3 Application Comparative Example 4 Application Comparative Example 5 D14 26.43 18.52 10.89 11.44 12.06 4.31 D28 33.31 22.67 16.78 15.93 14.57 5.67

[0078] From the above results, it can be seen that there is a significant difference in the amount of skin wrinkle volume reduction (cheek) in Application Example 1 compared with Application Comparative Examples 1 to 4, indicating that the composition for improving mitochondrial function in Example 2 can indirectly improve the skin condition, thereby having a good anti-wrinkle and anti-aging effect. Compared with Application Comparative Examples 1 to 4 and Application Comparative Example 5, there is still a significant difference, which may be because the improvement effect of mitochondrial membrane potential is only one of the factors affecting the amount of skin wrinkle volume reduction (cheek), and the total flavonoids extract of Prinus utilis and decapeptide-4 themselves also have good skin anti-wrinkle and anti-aging effects.

[0079] (3) Applicability to sensitive skin

[0080] During the testing period, the applicability rate of each application example for sensitive skin is shown in Table 7 below:

[0081] Table 7. Applicability of each application example to sensitive skin (unit: %)

[0082] Detection time Application Example 1 Application Comparative Example 1 Application Comparative Example 2 Application Comparative Example 3 Application Comparative Example 4 Application Comparative Example 5 D14 100 100 100 100 100 100 D28 100 100 100 100 100 100

[0083] From the above results, it can be seen that Application Example 1 and Application Comparative Examples 1 to 4 have excellent results in terms of the applicability rate for sensitive skin. This may be based on the fact that Application Comparative Example 5 itself is very gentle and very suitable for people with sensitive skin. Secondly, the composition for improving mitochondrial function in Example 2 itself is mild and non-irritating, and the total flavonoids extract of Prinus utilis and decapeptide-4 themselves are also very mild, and have a universal effect on sensitive skin.

[0084] The applicant declares that the above is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Those skilled in the art should understand that any changes or substitutions that can be easily thought of by those skilled in the art within the technical scope disclosed by the present invention are within the protection scope and disclosure scope of the present invention.

Claims

1. A composition for improving mitochondrial function, characterized in that: The invention comprises a total flavonoid extract of Prinus utilis and decapeptide-4 in a mass ratio of 100:1 to 125:

1.

2. The composition for improving mitochondrial function according to claim 1, characterized in that The total flavonoids extract of Prinus prinus is the extract of Prinus prinus seed meal after the Prinus prinus oil is removed.

3. The composition for improving mitochondrial function according to claim 2, characterized in that: The mass percentage of total flavonoids in the Prinus utilis total flavonoids extract is 30% to 60%.

4. The composition for improving mitochondrial function according to claim 3, characterized in that: The mass percentage of total flavonoids in the Prinus utilis total flavonoids extract is 56%.

5. The composition for improving mitochondrial function according to claim 1, characterized in that: The purity of the decapeptide-4 was 95%.

6. Use of a composition for improving mitochondrial function in cosmetics, characterized in that: The composition for improving mitochondrial function is the composition for improving mitochondrial function according to any one of claims 1 to 5.

7. Use of the composition for improving mitochondrial function in cosmetics according to claim 6, characterized in that: Cosmetics are any one of lotion, emulsion, essence, essence water, cream or mask.

8. Use of the composition for improving mitochondrial function in cosmetics according to claim 7, characterized in that: The lotion is in the form of a spray or an aqueous solution, and the essence water is in the form of a spray or an aqueous solution.

9. Use of the composition for improving mitochondrial function in cosmetics according to claim 7, characterized in that: The facial mask is in gel form.

10. An essence containing a composition for improving mitochondrial function, characterized in that: According to weight fraction, it includes the following components: water, 80.27 parts; Glycerin, 5 parts; Polydimethylsiloxane, 2 parts; Squalane, 1 part; 1,2-pentanediol, 3 parts; Bis-PEG-18 methyl ether dimethyl silane, 2 parts; Betaine, 2 parts; Methyl gluceth-20, 2 parts; Macrogol-8, 2 parts; Polyacrylate crosspolymer-6, 0.2 parts; p-Hydroxyacetophenone, 0.2 parts; Acrylates / C10-30 Alkyl Acrylate Crosspolymer, 0.1 parts; Sodium Hyaluronate, 0.05 parts; Disodium EDTA, 0.02 parts; Aminomethyl propanol, 0.06 parts; Total flavonoids extract of Prinus utilis, 0.1 part; 10 peptide-4, 0.001 parts.

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