A composition for skin barrier repair and anti-inflammation after non-surgical cosmetic treatments and its application.

Abstract

This invention belongs to the field of cosmetic technology, specifically relating to a composition for skin barrier repair and anti-inflammation after non-surgical cosmetic procedures and its application. The composition comprises Xinjiang comfrey peptide and Xinjiang comfrey naphthoquinone, with a mass ratio of 50-200:1. This composition synergistically inhibits the release of LPS-induced inflammatory factors, alleviating acute inflammatory reactions such as redness and burning after cosmetic procedures, achieving a dual effect of anti-inflammation and repair. The composition significantly reduces postoperative inflammatory redness and improves discomfort symptoms such as dryness and stinging. Through a multi-target mechanism, this invention solves the technical problems of existing repair products having single functions and slow onset of action, providing a highly efficient and comprehensive repair solution for skin after cosmetic procedures.

Description

Technical Field

[0001] This invention belongs to the field of cosmetic technology, specifically relating to a composition for skin barrier repair and anti-inflammation after non-surgical medical aesthetic treatments and its application. Background Technology

[0002] With the rapid development of medical aesthetic technology, non-surgical treatments such as ultrasound scalpels, lasers, and radiofrequency ablation have become widely popular due to their minimal invasiveness and rapid recovery. However, while these treatments target deep layers of the skin, they inevitably cause thermal damage or mechanical stimulation to the epidermis, leading to impaired skin barrier function. Clinical manifestations include inflammatory reactions such as redness, burning sensation, stinging, dryness, and peeling that appear immediately after the procedure or the following day. In severe cases, pigmentation or sensitization may even occur, affecting treatment outcomes and patient experience.

[0003] As a traditional Chinese medicine, the naphthoquinone compounds (such as shikonin) isolated from the roots of Xinjiang Lithospermum erythrorhizon have been proven to have good anti-inflammatory and wound-healing activities. However, the effects of single components on promoting cell migration and inhibiting inflammation are limited, making it difficult to achieve the ideal synergistic repair effect. There is an urgent need to develop a composition that can comprehensively utilize the different polar active ingredients of Xinjiang Lithospermum erythrorhizon and exert a synergistic effect. Summary of the Invention

[0004] The purpose of this section is to outline some aspects of the embodiments of the present invention and to briefly describe some preferred embodiments.

[0005] As one aspect of the present invention, the present invention provides a composition for skin barrier repair and anti-inflammation after non-surgical medical aesthetic treatment, wherein the composition is composed of Xinjiang comfrey peptide and Xinjiang comfrey naphthoquinone, and the mass ratio of Xinjiang comfrey peptide to Xinjiang comfrey naphthoquinone is 50-200:1. The preparation method of the Xinjiang Lithospermum peptide includes: using the residue after supercritical CO2 extraction of Xinjiang Lithospermum root as raw material, extracting with water to obtain an extract, adding Bacillus subtilis, neutral protease and aminopeptidase for enzymatic hydrolysis, and then ultrafiltration, nanofiltration and spray drying to obtain the final product; The preparation method of Xinjiang Lithospermum erythrorhizon includes: using Xinjiang Lithospermum erythrorhizon root as raw material, pre-treating it with high-pressure steam explosion, then performing supercritical CO2 extraction with ethanol as an entrainer, and purifying the resulting extract by molecular distillation.

[0006] As a preferred embodiment of the composition described in this invention, in the preparation method of the Xinjiang Lithospermum peptide, the water extraction is carried out by adding deionized water to the supercritical extraction residue of Xinjiang Lithospermum and extracting at 85-90℃ for 2-3 h; the Xinjiang Lithospermum peptide contains one or more peptides selected from the following: FN, PF, FA, FR, WG, QF, FYP, GPF, RF, FP, GQF, DGF, DPF, PPP, SPF, DW, FSP, PNF, FNP, FQP, GGPF, FDP, PDF, PGWPLFGH, PPPP, PGWPVFGH, PGGW.

[0007] As a preferred embodiment of the composition described in this invention, the residue after supercritical CO2 extraction of Xinjiang Lithospermum erythrorhizon root is obtained by using Xinjiang Lithospermum erythrorhizon root as raw material, pre-treating it with high-pressure steam explosion, and then performing supercritical CO2 extraction with ethanol as an entrainer, and collecting the residue after extraction.

[0008] As a preferred embodiment of the composition described in this invention, the preparation method of the Xinjiang purple gromwell peptide includes the following enzymatic hydrolysis: adjusting the temperature of the extract to 45-50℃, adjusting the pH to 7.5-8.0, adding Bacillus subtilis powder solution and neutral protease, and hydrolyzing for 2-2.5 hours; then adding aminopeptidase and hydrolyzing for 2-2.5 hours; and finally inactivating the enzyme after the hydrolysis is completed.

[0009] As a preferred embodiment of the composition described in this invention, the amount of Bacillus subtilis powder added is 2-3% of the volume of the extract, the amount of neutral protease added is 2-3% of the mass of the extract, and the amount of aminopeptidase added is 1-2% of the mass of the extract.

[0010] As a preferred embodiment of the composition described in this invention, in the preparation method of the Xinjiang purple gromwell peptide, the ultrafiltration is performed using a 5000 Da membrane core to collect the permeate; the nanofiltration is performed using a 200 Da membrane core to collect the concentrate; and hydroxypropyl methylcellulose is added before spray drying.

[0011] As a preferred embodiment of the composition described in this invention, in the preparation method of Xinjiang lithospermum naphthoquinone, the conditions for the high-pressure steam explosion pretreatment are a pressure of 0.8-1 MPa and a holding time of 20-30 seconds.

[0012] As a preferred embodiment of the composition described in this invention, in the method for preparing Xinjiang lithospermum naphthoquinone, the supercritical CO2 extraction conditions are: extraction temperature 40-45℃, extraction pressure 20-30 MPa, CO2 flow rate 40-200 L / h, multiple extractions are performed using ethanol as an entrainer, and the extracts are combined.

[0013] The present invention also provides the use of the composition in the preparation of skin care products for skin barrier repair and anti-inflammation after non-surgical medical aesthetic treatments, wherein the non-surgical medical aesthetic treatments include one or more of laser therapy, intense pulsed light therapy, radiofrequency therapy, ultrasound therapy, chemical peeling, and microneedling therapy.

[0014] Preferably, the composition is prepared as a cosmetic dosage form such as a cream, lotion, serum, or gel.

[0015] The beneficial effects of this invention are as follows: This invention achieves a significant synergistic effect by specifically combining Xinjiang lithospermum peptide and Xinjiang lithospermum naphthoquinone. The combination significantly promotes the migration ability of keratinocytes, accelerates the re-epithelialization process of the wound, and thus rapidly repairs the damaged skin barrier. The composition of this invention can synergistically inhibit the release of LPS-induced inflammatory factors, alleviate acute inflammatory reactions such as redness and burning after cosmetic procedures, and achieve dual anti-inflammatory and repairing effects. The composition of this invention can significantly reduce postoperative inflammatory red areas and improve discomfort symptoms such as dryness and stinging. Through a multi-target mechanism, this invention solves the technical problems of existing repair products having single functions and slow onset of action, providing a highly efficient and comprehensive repair solution for post-cosmetic procedures. Attached Figure Description

[0016] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below, wherein: Figure 1 This is a diagram of the cell migration experiment in Example 1. Detailed Implementation

[0017] To make the above-mentioned objectives, features and advantages of the present invention more apparent and understandable, the specific embodiments of the present invention will be described in detail below with reference to specific examples.

[0018] Experimental materials: Peptone, catalog number 01-001, Beijing Aoboxing Biotechnology Co., Ltd.; D-anhydrous glucose, Beijing Bio-Top Technology Co., Ltd.; Yeast extract, Beijing Solarbio Technology Co., Ltd.; Neutral protease, CAS No. 9068-59-1, Beijing Bio-Top Technology Co., Ltd.; Aminopeptidase, Cangzhou Xiasheng Enzyme Biotechnology Co., Ltd.; Hydroxypropyl methylcellulose, type 2910, Shandong Liaocheng Ahua Pharmaceutical Co., Ltd.

[0019] The supercritical CO2 extraction instrument, model HA220-50-06-C, was purchased from Nantong Huaan Supercritical Extraction Co., Ltd.; the molecular distillation instrument, model QYMD-80C, was purchased from Qiyu Industrial (Shanghai) Co., Ltd. The QB-200 steam explosion test bench was manufactured by Henan Hebi Zhengdao Heavy Machinery Factory.

[0020] Preparation method of Xinjiang lithosine: Step 1: Pretreatment: Crush the dried Xinjiang purple gromwell root into powder using a pulverizer, sieve (0.355mm aperture), retain the sieved portion, and load it into a high-pressure steam explosion reactor. Use high-pressure instantaneous explosion technology to maintain pressure at 0.8 MPa for 30 seconds. After the pressure holding is completed, open the explosion valve to rupture the cell walls and release the active ingredients, thus obtaining the pretreated material.

[0021] Step 2: Supercritical CO2 Extraction: 10 kg of the material obtained in Step 1 was loaded into the extraction vessel, and 10 kg of ethanol was added. The mixture was stirred and stirred until homogeneous. The extraction vessel was heated to 45 °C, the separation vessel to 35 °C, and the storage tank to 30 °C. Then, the CO2 cylinder was turned on, and the CO2 flow rate was controlled at 160 L / h. The system was pressurized using a high-pressure pump. When the pressure in the extraction vessel was 30 MPa and the pressure in the separation vessel was 6 MPa, the circulation extraction began. The extraction vessel was kept at a constant temperature and pressure. After 2 hours of extraction, the material was discharged from the outlet of the separation vessel, completing the first extraction. After the first extraction, 10 kg of ethanol was pumped in using an entrainer pump, and the above experimental operation was repeated. After 2 hours of extraction, the material was discharged from the outlet of the separation vessel, completing the second extraction. After the second extraction, 10 kg of ethanol was pumped in using an entrainer pump, and the above experimental operation was repeated. After 2 hours of extraction, the pink extract of *Lithospermum erythrorhizon* was obtained from the third extraction. The extract solutions from the three extractions were combined to obtain the crude total naphthoquinone extract solution of *Lithospermum erythrorhizon*.

[0022] Step 3: Molecular distillation: Take the crude total naphthoquinone extract solution from Xinjiang Lithospermum obtained in Step 2, and set the molecular distillation instrument parameters as follows: feed temperature 50 ℃, evaporator temperature 80 ℃, condenser -5 ℃, external condenser -20 ℃, rotor speed 300 rpm. Pump the feed solution into the molecular distillation instrument, turn on heating, condensation, vacuuming, and stirring, discard the light components, and collect the heavy component samples. Concentrate the solution in a rotary evaporator at 50 ℃ to obtain the naphthoquinone extract from Xinjiang Lithospermum.

[0023] Preparation method of Xinjiang Lithospermum erythrorhizon peptide: (1) Preparation of supercritical extraction residue of Lithospermum erythrorhizon from Xinjiang: Pretreatment: The dried Xinjiang purple gromwell root is pulverized into powder using a pulverizer and sieved (0.355 mm aperture). The sieved portion is retained and loaded into a high-pressure steam explosion reactor. High-pressure instantaneous explosion technology is used to maintain the pressure at 0.8 MPa for 30 seconds. After the pressure is maintained, the explosion valve is opened to rupture the cell walls and release the active ingredients, thus obtaining the pretreated material.

[0024] Supercritical CO2 extraction: 1 kg of pretreated material was loaded into the extraction vessel, and 0.5 kg of GTCC was added and stirred until homogeneous. The extraction vessel was heated to 45 ℃, the separation vessel to 45 ℃, and the storage tank to 35 ℃. Then, the CO2 cylinder was turned on, and the CO2 flow rate was controlled at 50 L / h. The system was pressurized by a high-pressure pump. When the pressure in the extraction vessel was 40 MPa and the pressure in the separation vessel was 5 MPa, the circulation extraction began, and the extraction vessel was kept at a constant temperature and pressure. After extraction for 1 h, the material was discharged from the outlet of the separation vessel, the residue was retained, and the extract was discarded. After the first extraction was completed, 1 kg of GTCC was pumped in through an entrainer pump, and the above experimental operation was repeated. The second extraction was completed after 1 h, the residue was retained, and the extract was discarded. After the second extraction was completed, 1 kg of GTCC was pumped in through an entrainer pump, and the above experimental operation was repeated. The residue was retained, the extract was discarded, and the residues were combined to obtain the supercritical extraction residue of Xinjiang Lithospermum.

[0025] (2) Take 100g of Xinjiang Lithospermum supercritical extraction residue and add 3000mL of deionized water, extract at 90℃ for 2h.

[0026] (3) Preparation of Bacillus subtilis powder solution: Dissolve 3g of Bacillus subtilis powder (purchased from Henan Xinyangshao Biotechnology Co., Ltd., batch number KC20250609) in 1kg of culture medium; the culture medium composition is: peptone 10g / L, glucose 10g / L, yeast powder 5g / L, and deionized water as solvent; the mass concentration of Bacillus subtilis powder solution is 0.3%.

[0027] (4) Adjust the temperature of the extract obtained in step (2) to 45℃ and the pH to 7.5. Add 2% (volume percentage) of Bacillus subtilis powder solution prepared in step (3), and add 2wt% of neutral protease. Enzymatically hydrolyze for 2h, then add 1wt% of aminopeptidase and enzymatically hydrolyze for 2h. After the enzymatic hydrolysis is completed, keep at 90℃ for 10min to inactivate the enzyme.

[0028] (5) After cooling to room temperature, filter using a 0.45 μm filter plate. Then perform ultrafiltration using a 5000 Da membrane element and collect the permeate. Then replace with a 200 Da membrane element for nanofiltration and collect the concentrate. Concentrate the liquid using a rotary evaporator at 60℃ to 500 mL; add 0.8% hydroxypropyl methylcellulose to the liquid, set the inlet air temperature to 150℃, the outlet air temperature to 85℃, and the flow rate to 200 mL / h, and spray dry to obtain Xinjiang Lithospermum erythrorhizon peptide extract.

[0029] The Xinjiang lithospermum peptide obtained from the extract was docked with TRPV4 molecules to determine its activity. The predicted binding energy ranking of Xinjiang lithospermum peptide and TRPV4 is as follows: MM > GF > SF > FG > AF > DF (ΔG = -6.834) > FN (ΔG = -7.113) > PF > FA > FR > WG > QF > FYP > GPF > RF > FP > GQF > DGF > DPF > PPP > SPF > DW > FSP > PNF > FNP > FQP > GGPF > FDP > PDF > PGWPLFGH (ΔG = -8.721) > PPPP (ΔG = -9.002) > PGWPVFGH > PGGW. The amino acid sequences with four or more amino acids are shown in SEQ ID NO: 1-5.

[0030] Example 1: Human immortalized keratinocytes (HaCaT) in the logarithmic growth phase were harvested, digested, and resuspended. The cells were seeded into 6-well cells at the number expected to fully colonize the culture plates on day 2, with three replicates per group. Once the cells had fully colonized the plates, a scratching technique was used to create uniform scratches in the cells, simulating wound formation. The medium was then replaced with serum-free medium, and different concentrations of the drug were added for treatment. Cell migration was observed under a microscope at 0 h and 24 h after scratching, and the migration rate and relative migration rate were calculated for each group.

[0031] The formula for calculating mobility (MR) is: MR = (S0 - S) 24 ) / S0×100%.

[0032] in: S0 is the area of ​​the scratched region at 0h; S 24 The area of ​​the scratched region after 24 hours; the formula for calculating the relative migration rate (RMR) is: RMR%=MR 样品组 / MR 空白组 ×100%.

[0033] Preparation of Xinjiang Lithospermum peptide solution: Take dried Xinjiang Lithospermum peptide powder, add it to high-sugar DMEM medium, vortex for 1 min, and prepare a 1 mg / mL Xinjiang Lithospermum peptide stock solution.

[0034] Preparation of Xinjiang lithonaquinone solution: Take dried Xinjiang lithonaquinone powder, add DMSO solution, vortex for 1 min, and prepare a 1 mg / mL Xinjiang lithonaquinone stock solution.

[0035] During the experiment, take an appropriate amount of the stock solution according to the required concentration and add it to serum-free DMEM medium.

[0036] Determine whether there is a synergistic effect using the formula of the Bliss independent model: The formula for calculating the Bliss independent model (Bliss co-factor) is: ΔE = Eab - (Ea + Eb - Ea × Eb). Where: Eab: Relative migration rate improvement of the combination group; Ea, Eb: Relative mobility improvement values ​​of the single addition group; If ΔE>0, it indicates that there is a synergistic effect between the samples.

[0037] Based on the concentrations of the main active ingredient (Xinjiang lithospermum peptide) in the Xinjiang lithospermum peptide extract and the main active ingredient (Xinjiang lithospermum naphthoquinone) in the Xinjiang lithospermum naphthoquinone extract, the migration rate and relative migration rate of each group were calculated according to the experimental concentrations in Table 1.

[0038] Table 1

[0039] Furthermore, compared to using Xinjiang shikonin alone, its combination with Xinjiang shikonin significantly enhances cell migration promotion. According to the Bliss independent model, the synergistic values ​​ΔE for the 5 μg / ml Xinjiang shikonin + 0.1 μg / ml Xinjiang shikonin and 20 μg / ml Xinjiang shikonin + 0.1 μg / ml Xinjiang shikonin groups were 0.40 and 0.47, respectively, indicating that the combination of Xinjiang shikonin and Xinjiang shikonin synergistically promotes cell migration, and the optimal ratio of the two is 50:1-200:1.

[0040] Example 2: Human immortalized keratinocytes (HaCaT) in the logarithmic growth phase were harvested, digested, resuspended, and seeded into 24-well cell culture plates, with three replicates per group. When the cells reached approximately 50% confluence, culture medium containing 5 μg / ml LPS and different samples was added. After culturing for another 24 hours, the supernatant was collected, and the TNF-α level in each group was detected by ELISA. The Bliss independent model calculation formula was the same as in Example 1.

[0041] Table 2

[0042] For LPS-induced TNFα, both Xinjiang lithospermum peptide and Xinjiang lithospermum naphthoquinone can effectively inhibit its secretion.

[0043] Furthermore, compared to using Xinjiang shikonin alone, its combination with Xinjiang shikonin significantly inhibited TNFα secretion. According to the Bliss independent model, the synergistic values ​​ΔE for the 5 μg / ml Xinjiang shikonin + 0.1 μg / ml Xinjiang shikonin and 20 μg / ml Xinjiang shikonin + 0.1 μg / ml Xinjiang shikonin groups were 0.15 and 0.24, respectively, indicating that the combination of Xinjiang shikonin and Xinjiang shikonin can synergistically inhibit TNFα secretion, thereby exerting a good anti-inflammatory and soothing effect. The optimal combination ratio is 50:1-200:1.

[0044] Example 3: One hundred volunteers, aged 25-57 years (mean age 42.13±8.91 years), were recruited as study subjects. They first underwent Peninsula M4.5 and M3.0 ultrasonic scalpel treatment, followed by Peninsula D4.5 and D3.0 ultrasonic beam therapy. Twenty-four subjects who experienced adverse reactions such as redness, swelling, pain, dryness, and peeling on the second day were randomly divided into four groups. Group ① used a base cream without Xinjiang purple gromwell peptide and Xinjiang purple gromwell naphthoquinone (control group); Group ② used a repair cream containing the aforementioned Xinjiang purple gromwell peptide and Xinjiang purple gromwell naphthoquinone combination; Group ③ used a cream containing an equal amount of Xinjiang purple gromwell peptide; and Group ④ used a cream containing an equal amount of Xinjiang purple gromwell naphthoquinone. Subjects applied the cream to the affected area 2-3 times daily. Facial redness was recorded using VISA before and after one week of continuous use.

[0045] Comparison of inflammatory red areas: Based on the VISIA skin testing instrument and clinical experience, the efficacy criteria were defined as follows: a reduction of more than 60% in the red area of ​​the treated patient was considered a basic cure; a reduction of 30%-59% was considered significant effect; a reduction of 10%-29% was considered effective; and a reduction of <10% was considered ineffective. One week after using each group of samples, photos taken before and after treatment were compared to calculate the reduction rate of the inflammatory red area in each group.

[0046] The main components of the face cream composition are shown in Table 3, and the specific preparation process is as follows: 1. Raw material pretreatment: Add Xinjiang shikonin to caprylic / capric triglyceride to prepare a 0.1 wt% Xinjiang shikonin solution (hereinafter referred to as Xinjiang shikonin oil), and then add 5 wt% Xinjiang shikonin oil to the formula.

[0047] 2. Preparation of aqueous phase: Mix components 1 and 2 as the aqueous phase, stir at low speed for 1 min at room temperature (25℃) until completely mixed; add 3, and continue stirring at low speed for 2 min until uniformly dispersed.

[0048] 3. Oil phase preparation: Mix components 4, 5, 6 and 7 as the oil phase, heat at low temperature to 55-60℃, stir at low speed for 3 minutes until the raw materials melt and are evenly mixed.

[0049] 4. Emulsification: Slowly and evenly pour the preheated oil phase into the water phase while stirring, maintaining the temperature at 55-60℃, and stir rapidly for 8 minutes until the system is a fine and thin cream. Stop heating and let it cool naturally to below 30℃ while stirring at low speed. Add 0.5% phenoxyethanol and stir for 2 minutes until homogeneous.

[0050] Table 3

[0051] Table 4

[0052] Conclusion: The experimental results show that the overall effective rate of the face cream using the combination of Xinjiang purple gromwell peptide and Xinjiang purple gromwell naphthoquinone is significantly higher than that of the face cream using Xinjiang purple gromwell peptide alone or Xinjiang purple gromwell naphthoquinone alone.

[0053] Comparative Example 1: (1) The dried Xinjiang purple gromwell root was pulverized into powder by a pulverizer and sieved (0.355 mm aperture). The sieved part was retained. 100g of powder was added to 3000mL of deionized water and extracted at 90℃ for 2h.

[0054] (2) Adjust the temperature to 45℃, adjust the pH to 7.5, add 2% neutral protease, and enzymatically hydrolyze for 2h; then add 1% aminopeptidase and enzymatically hydrolyze for 2h; after the enzymatic hydrolysis is completed, keep at 90℃ for 10min to inactivate the enzyme.

[0055] (3) After cooling to room temperature, filter with a 0.45μm filter plate, then use a 5000Da membrane core for ultrafiltration, collect the permeate, then replace with a 200Da membrane core for nanofiltration, and collect the concentrate.

[0056] (4) Use a rotary evaporator to concentrate the liquid at 60°C to 500mL, add 0.8% hydroxypropyl methylcellulose, set the inlet air temperature to 150°C, the outlet air temperature to 85°C, and the flow rate to 200mL / h for spray drying.

[0057] Comparative Example 2: (1) Take 100g of supercritical extraction residue of Lithospermum erythrorhizon from Xinjiang and mix it with 10g of water. Put it into a 2500ml conical flask, stir it evenly, seal it with plastic wrap, sterilize it at 121℃ for 20 minutes, and obtain solid fermentation culture medium after cooling. Inoculate it with Bacillus subtilis and place it in a constant temperature incubator at 28℃ for solid fermentation for 3 days. Finally, sterilize it at 121℃ for 20 minutes.

[0058] (2) Take 100g of fermented powder and add it to 3000mL of deionized water, and cook at 90℃ for 2h.

[0059] (3) Adjust the temperature to 45℃, adjust the pH to 7.5, add 2% neutral protease, and enzymatically hydrolyze for 2h; then add 1% aminopeptidase and enzymatically hydrolyze for 2h; after the enzymatic hydrolysis is completed, keep at 90℃ for 10min to inactivate the enzyme.

[0060] (4) After cooling to room temperature, filter using a 0.45 μm filter plate. Then perform ultrafiltration using a 5000 Da membrane element and collect the permeate. Then replace with a 200 Da membrane element for nanofiltration and collect the concentrate.

[0061] (5) Use a rotary evaporator to concentrate the liquid at 60°C to 500mL; add 0.8% hydroxypropyl methylcellulose to the liquid, set the inlet air temperature to 150°C, the outlet air temperature to 85°C, and the flow rate to 200mL / h for spray drying.

[0062] Polypeptide content determination experiment (biuret reagent method): 1) Biuret reagent: Reagent A: 0.15g CuSO4, 0.6g potassium sodium tartrate, and 50mL distilled water are mixed evenly; Reagent B: 30mL 10% NaOH solution (mix A and B evenly before use).

[0063] 2) Preparation of bovine serum albumin standard curve: (Standard range 0-5 mg / mL) First, prepare a 10 mg / mL bovine serum albumin solution (water as solvent). Take 0, 50, 100, 150, 200, and 250 μL into EP tubes respectively, add water to 0.5 mL, add 2 mL of biuret reagent (A and B mixed before use), react at room temperature for 30 min, and measure its absorbance at 540 nm to prepare the standard curve.

[0064] 3) Determination of sample solution: Take the sample, add 1.0 mL of 10% trichloroacetic acid to dissolve it, centrifuge to precipitate the protein, take 0.5 mL of supernatant, add 2 mL of biuret reagent (A and B mixed before use), react at room temperature for 30 min, measure its absorbance at 540 nm, and calculate the sample peptide content according to the standard curve.

[0065] 4) Formula for calculating polypeptide content: The standard curve for polypeptide content is y = Ax + B(R). 2 >0.99); Polypeptide content (%) = protein or polypeptide concentration (mg / mL) / sample solution concentration (mg / mL) × 100%. The experimental results are shown in Table 5.

[0066] Table 5

[0067] In vitro antioxidant effect experiment (DPPH free radical inhibition experiment): Preparation of DPPH ethanol solution: Weigh 20 mg of DPPH, dissolve it in anhydrous ethanol, and dilute to a final volume of 250 mL in a volumetric flask. The DPPH concentration is prepared to be 2 × 10⁻⁶ mg / mL. -4 mol / L; store protected from light at 0-4 ℃, prepare and use immediately, effective within 4 hours. (Positive control: Vitamin C: 1 mg / mL).

[0068] Experimental steps: 1) Take 1 mL of the test solution and 1 mL of 2×10⁻⁶ solution. -4 Mix the mol / L DPPH solution thoroughly (tube A). 2) Take 1 mL of solvent and 1 mL of 2×10 -4 Mix the mol / L DPPH solution thoroughly (tube B). 3) Mix 1 mL of solvent with 1 mL of the test solution (tube C). 4) After reacting in the dark for 30 minutes, measure the absorbance values ​​of tubes A, B, and C at 517 nm.

[0069] Table 6 Reagent Proportioning Table

[0070] Formula for calculating DPPH free radical inhibition rate: DPPH inhibition rate (%) = (B + CA) / B × 100%.

[0071] The results of the DPPH free radical inhibition experiment are shown in Table 7.

[0072] Table 7 Results of DPPH free radical scavenging rate determination

[0073] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications or substitutions should be covered within the scope of the claims of the present invention.

Claims

1. A composition for skin barrier repair and anti-inflammation after non-surgical cosmetic treatments, characterized in that, The composition consists of Xinjiang lithospermum peptide and Xinjiang lithospermum naphthoquinone, wherein the mass ratio of Xinjiang lithospermum peptide to Xinjiang lithospermum naphthoquinone is 50-200:

1. The preparation method of the Xinjiang Lithospermum peptide includes: using the residue after supercritical CO2 extraction of Xinjiang Lithospermum root as raw material, extracting with water to obtain an extract, adding Bacillus subtilis, neutral protease and aminopeptidase for enzymatic hydrolysis, and then ultrafiltration, nanofiltration and spray drying to obtain the final product; The preparation method of Xinjiang Lithospermum erythrorhizon includes: using Xinjiang Lithospermum erythrorhizon root as raw material, pre-treating it with high-pressure steam explosion, then performing supercritical CO2 extraction with ethanol as an entrainer, and purifying the resulting extract by molecular distillation.

2. The composition according to claim 1, characterized in that, In the preparation method of the Xinjiang Lithospermum peptide, the water extraction is carried out by adding deionized water to the supercritical extraction residue of Xinjiang Lithospermum and extracting at 85-90℃ for 2-3 h; the Xinjiang Lithospermum peptide contains one or more peptides selected from the following peptides: FN, PF, FA, FR, WG, QF, FYP, GPF, RF, FP, GQF, DGF, DPF, PPP, SPF, DW, FSP, PNF, FNP, FQP, GGPF, FDP, PDF, PGWPLFGH, PPPP, PGWPVFGH, PGGW.

3. The composition according to claim 1, characterized in that, The residue after supercritical CO2 extraction of Xinjiang Lithospermum erythrorhizon root is obtained by using Xinjiang Lithospermum erythrorhizon root as raw material, pre-treating it with high-pressure steam explosion, and then performing supercritical CO2 extraction with ethanol as entrainer, and collecting the residue after extraction.

4. The composition according to claim 2, characterized in that, In the preparation method of Xinjiang purple gromwell peptide, the enzymatic hydrolysis includes: adjusting the temperature of the extract to 45-50℃, adjusting the pH to 7.5-8.0, adding Bacillus subtilis powder solution and neutral protease, and hydrolyzing for 2-2.5 hours; then adding aminopeptidase and hydrolyzing for 2-2.5 hours; and inactivating the enzyme after the hydrolysis is completed.

5. The composition according to claim 4, characterized in that, The amount of Bacillus subtilis powder added is 2-3% of the volume of the extract, the amount of neutral protease added is 2-3% of the mass of the extract, and the amount of aminopeptidase added is 1-2% of the mass of the extract.

6. The composition according to any one of claims 1-5, characterized in that, In the preparation method of Xinjiang purple gromwell peptide, the ultrafiltration is performed using a 5000Da membrane core to collect the permeate; the nanofiltration is performed using a 200Da membrane core to collect the concentrate; and hydroxypropyl methylcellulose is added before spray drying.

7. The composition according to any one of claims 1-5, characterized in that, In the preparation method of Xinjiang lithospermum naphthoquinone, the conditions for the high-pressure steam explosion pretreatment are a pressure of 0.8-1 MPa and a holding time of 20-30 seconds.

8. The composition according to any one of claims 1-5, characterized in that, In the preparation method of Xinjiang lithospermum naphthoquinone, the supercritical CO2 extraction conditions are: extraction temperature 40-45℃, extraction pressure 20-30 MPa, CO2 flow rate 40-200 L / h, multiple extractions are performed using ethanol as an entrainer, and the extracts are combined.

9. The use of the composition according to claim 1 in the preparation of skin care products for skin barrier repair and anti-inflammation after non-surgical medical aesthetic treatments, characterized in that, The non-surgical cosmetic treatments include one or more of the following: laser therapy, intense pulsed light therapy, radiofrequency therapy, ultrasound therapy, chemical peels, and microneedling therapy.

10. The application according to claim 9, characterized in that, The composition is prepared as a cosmetic dosage form such as a cream, lotion, serum, or gel.

Images