Rhizoma picrorhizae extract and function and application thereof

Abstract

The invention relates to the field of cosmetic application and development, in particular to a rhizoma picrorhizae extract and functions and application thereof. The rhizoma picrorhizae extract contains picroside I and picroside II, and the mass ratio of the picroside I to the picroside II is (2-7): 7. According to the invention, the rhizoma picrorhizae extract can significantly reduce skin grease and skin oil and gloss, and can also have anti-inflammatory, soothing, radiation-proof and anti-aging functions on the skin.

Description

Technical Field

[0001] The invention relates to the field of cosmetic application development, and in particular to a picrorhizome extract and its function and use. Background Art

[0002] Picrorhiza chinensis is the dried rhizome of Neopicrorhiza scrophulariiflora (Pennell)DYHong, a plant of the Plantaginaceae family. It was first discovered in India and Indonesia. In the 1970s, its close relative, wild picrorhiza chinensis, was discovered in my country and became its substitute. It is mainly distributed in the high-altitude mountainous areas of 3600-4400m in Sichuan, northwest Yunnan and southern Tibet, and has been included in the "Pharmacopoeia of the People's Republic of China" of all dynasties. Domestic and foreign research results show that the chemical components of picrorhiza chinensis mainly include cyclopentadiene ether terpenes, phenylethanol glycosides, and phenolic glycosides, among which cyclopentadiene ether terpenes are the main active ingredients of picrorhiza chinensis. Modern pharmacological studies have shown that it has liver protection, anti-inflammatory, neuroprotective, and anti-tumor effects. Its extracts are often used in the study of pharmacologically related diseases, such as the treatment of hepatitis, urinary tract infections, and anti-liver damage. At present, there are few studies on the application of picrorhiza chinensis or its extracts in cosmetics.

[0003] Therefore, there is an urgent need in this field to conduct in-depth research on the application possibilities of Picrorhizoma Coptidis or its extracts. Summary of the invention

[0004] The present invention aims to conduct an in-depth study on the application possibility of picrorhizoma berberis or its extract, and specifically provides a new function and use of a picrorhizoma berberis extract.

[0005] In a first aspect of the present invention, a picroside extract is provided, wherein the picroside extract contains picroside I and picroside II, and the mass ratio of the picroside I to the picroside II is 2-7:7.

[0006] In another preferred embodiment, the mass ratio of picroside I to picroside II in the Picrorhizoma Coptidis extract is 2.8-7:7.

[0007] In another preferred embodiment, the mass ratio of picroside I to picroside II in the Picrorhizoma Coptidis extract is 3-4:7.

[0008] In another preferred embodiment, the peak area ratio of picroside I to picroside II in the Picrorhizoma Coptidis extract is 5.5-8:7.

[0009] In another preferred embodiment, the content of picroside I and picroside II is 40%-60%, based on the total mass of the Picrorhizon extract.

[0010] In another preferred example, the content of picroside I and picroside II is 45%-55%, based on the total mass of the Picrorhizon extract.

[0011] In another preferred embodiment, the method comprises the steps of: (1) Extraction: providing a mixture of Picrorhizoma Coptidis raw materials and ethanol, and performing reflux extraction for 1-3 times to obtain a crude extract; (2) Concentration: Concentrating the crude extract obtained in step (1) under reduced pressure to obtain a crude extract concentrate; (3) Resin purification: The crude extract obtained in step (2) is loaded onto a macroporous resin, first eluted with water and then with ethanol, the ethanol eluate is collected, and concentrated under reduced pressure to obtain a Picrorhizoma Coptidis extract; (4) Freeze-drying: freeze-drying the picrorhizoma picrorhizoma extract obtained in step (3) to prepare lyophilized powder of picrorhizoma picrorhizoma extract, i.e., the picrorhizoma picrorhizoma extract.

[0012] In another preferred embodiment, the concentration of ethanol in step (1) is 65 v / v%-75 v / v%.

[0013] In another preferred embodiment, two reflux extractions are performed in step (1), and the solid-liquid ratio of the first reflux extraction is 1:9-11; and the solid-liquid ratio of the second reflux extraction is 1:7-9.

[0014] In another preferred embodiment, the reflux temperature in step (1) is 80-90°C.

[0015] In another preferred embodiment, the concentration temperature in step (2) is 55-60° C., and the vacuum degree is -0.09 MPa to -0.07 MPa.

[0016] In another preferred embodiment, the reflux extraction in step (1) is performed twice, the solid-liquid ratio of the first reflux extraction is 1:9-11, the heating temperature is 80-90°C, and the extraction time is 1.5-2.5h; the solid-liquid ratio of the second reflux extraction is 1:7-9, the heating temperature is 80-90°C, and the heating time is 0.5-1.5h; and filtering is performed using filter cloth to obtain a crude extract.

[0017] In another preferred embodiment, the concentration temperature in step (2) is 55-60°C, the vacuum degree is -0.09MPa~-0.07MPa, and the concentration is 1.5-2.5 times of the feed amount.

[0018] In another preferred embodiment, the macroporous resin in step (3) is selected from the following group: HP20, AB-8, D101 macroporous resin.

[0019] In another preferred embodiment, the macroporous resin in step (3) is HP20.

[0020] In another preferred embodiment, the elution volume of the water elution in step (3) is 2.5-3.5 BV. In another preferred embodiment, the ethanol concentration of the ethanol elution in step (3) is 65 v / v%-75% v / v ethanol.

[0021] In another preferred embodiment, the elution volume of the ethanol elution in step (3) is 2.5-3.5 BV.

[0022] In another preferred embodiment, the freeze-drying time in step (4) is ≥ 24 h.

[0023] In another preferred embodiment, the freezing temperature of the freeze-drying in step (4) is -70°C to -50°C.

[0024] In another preferred embodiment, the vacuum degree of freeze-drying in step (4) is ≤20 Pa.

[0025] In the second aspect of the present invention, there is provided a use of the Picrorhizoma Coptidis extract as described in the first aspect of the present invention, for preparing a composition for lowering the oil content of the skin.

[0026] In another preferred embodiment, the composition reduces the skin oil content by ≥20%.

[0027] In another preferred embodiment, the skin oil is secreted by sebaceous gland cells.

[0028] In another preferred embodiment, the composition is used to reduce skin oiliness.

[0029] In another preferred embodiment, the composition is also used to regulate the PPARγ signaling pathway, thereby affecting lipid synthesis.

[0030] In another preferred embodiment, the composition reduces FABP4 gene expression.

[0031] In another preferred embodiment, the composition reduces the expression of PPARGC1A gene.

[0032] In another preferred embodiment, the composition reduces the expression of genes related to inflammation.

[0033] In another preferred embodiment, the composition reduces the expression of inflammation-related genes FOS, TNF, and IL21R.

[0034] In another preferred embodiment, the composition reduces the expression of matrix metalloproteins associated with aging.

[0035] In another preferred embodiment, the composition reduces the expression of matrix metalloproteins MMP1 and MMP10 associated with aging.

[0036] In another preferred embodiment, the composition is also used to reduce the level of active oxygen.

[0037] In a third aspect of the present invention, a composition is provided, comprising: (1) the Picrorhizoma Coptidis extract as described in the first aspect of the present invention; and (2) additional ingredients that can be used in daily chemical products.

[0038] In another preferred embodiment, the composition is used for a purpose selected from the group consisting of oil control, anti-inflammatory and soothing, radiation protection, anti-aging, or a combination thereof.

[0039] In another preferred embodiment, the composition has one or more characteristics selected from the following group: (1) Reduce skin oil content; (2) reduce FABP4 gene expression; (3) reduce PPARGC1A gene expression; (4) reducing the expression of inflammation-related genes; preferably, the inflammation-related genes are selected from the group consisting of FOS, TNF, and IL21R genes; (5) reducing the expression of matrix metalloproteins associated with aging; preferably, the matrix metalloproteins associated with aging are selected from the group consisting of: MMP1, MMP10; (6) Reduce the level of reactive oxygen species.

[0040] In another preferred embodiment, the composition is a cream or an emulsion.

[0041] In another preferred embodiment, the composition is selected from the following group: facial cream, skin care water, isolation cream, lotion, facial mask, shampoo, and conditioner.

[0042] In another preferred embodiment, the content of picroside I in the composition is ≥10 μg / g.

[0043] In another preferred embodiment, the content of picroside II in the composition is ≥10 μg / g.

[0044] In another preferred embodiment, the mass ratio of picroside I to picroside II in the composition is 2-7:7, preferably 2-4:7.

[0045] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 The HPLC profile of Picrorhizoma Coptidis extract is shown.

[0047] Figure 2 Examples of the applied area and the control area in the short-term oil control experiment are shown.

[0048] Figure 3 Shows the results of a short-term oil control experiment.

[0049] Figure 4 Shows the results of a long-term oil control experiment.

[0050] Figure 5 Shows comparison photos of two volunteers in a long-term oil control experiment using the test samples at 0 days and 28 days.

[0051] Figure 6 The transcriptomic KEGG enrichment of Picrorhizoma extract is shown.

[0052] Figure 7 The results show that the picrorhizon extract has an effect on the expression of FABP4 gene and PPARGC1A gene. Figure 7 A in the figure is the expression of the fatty acid binding protein FABP4 gene in the lipid metabolism PPAR signaling pathway. Figure 7 B in the figure shows the expression of PPARγ coactivator PPARGC1A gene.

[0053] Figure 8 The effect of Picrorhizoma Coptidis extract on the expression of genes related to inflammatory factors is shown.

[0054] Fig. 9 The effect of Picrorhizoma Coptidis extract on the expression of aging factor matrix metalloproteinase was shown.

[0055] Fig.10 The relative viability of cells at different sample concentrations is shown.

[0056] Fig.11 The results show that the lyophilized powder of Picrorhizoma Coptidis extract can inhibit ROS secretion.

[0057] Fig.12 The results show that the lyophilized powder of Picrorhizoma Coptidis extract inhibits the secretion of TNF-α.

[0058] Fig.13 The results show that the lyophilized powder of Picrorhizoma Coptidis extract inhibits IL-6 secretion. DETAILED DESCRIPTION

[0059] After extensive and in-depth research and a large number of experimental screening, the inventors unexpectedly discovered for the first time that the picrorhizon extract has the functions of oil control, anti-inflammatory and soothing, radiation protection, and anti-aging, and can be used to prepare products containing the picrorhizon extract. The mass ratio of picrorhizon glycoside I and picrorhizon glycoside II in the picrorhizon extract is 2-7:7. In the present invention, the oil control effect of the picrice extract is evaluated by human efficacy test, and the results show that the picrice extract can significantly reduce skin oil and skin oiliness, indicating that the picrice extract has obvious oil control effect and can be used as an effective ingredient in daily chemical products; the oil control mechanism of the picrice extract is analyzed by transcriptomics, showing that the picrice extract affects lipid synthesis by significantly inhibiting the expression of fatty acid binding protein FABP4 gene in lipid metabolism PPAR signaling pathway, and finally plays a role in reducing skin oil secretion, providing a scientific basis at the gene level for the application of the picrice extract in oil control daily chemical products; by evaluating the efficacy of the picrice extract in reducing the level of active oxygen, reducing the damage of ultraviolet rays to cells, and reducing the secretion of inflammatory factors, it is shown that the picrice extract has anti-inflammatory, soothing, radiation protection, and anti-aging functions on the skin. Therefore, the picrice extract provides a new idea for the application of daily chemical products of oil control, anti-inflammatory, soothing, radiation protection, and anti-aging. On this basis, the inventors have completed the present invention.

[0060] Terminology Picrorhizon Extract Wild picrorhizoma rapae is a national second-class protected species, native to Tibet and northwest Yunnan at an altitude of 3900-4500m, with a narrow distribution area. Due to long-term over-exploitation, wild resources are on the verge of extinction. At present, picrorhizoma rapae has been cultivated on a large scale in Yunnan. The present invention expands the application of picrorhizoma rapae extract in daily chemical products, promotes the healthy development of picrorhizoma planting industry, and realizes the protection and sustainable utilization of endangered medicinal plant resources.

[0061] Uses of Picrorhizoma Extract Sebaceous gland cells: Sebaceous glands are multicellular glands that secrete sebum from the openings of the sebaceous glands that open on the inner surface of the pores. Sebaceous gland cells exist in the sebaceous glands and play a role in releasing sebum. The secreted sebum protects and moisturizes the skin and hair. Abnormal secretion of sebum is the cause of skin diseases such as oily skin, dry skin, acne, seborrhea, seborrheic dermatitis, and xeroderma.

[0062] In the present invention, the oil control effect of the picrorhizon extract is evaluated by human efficacy test, and the test formula containing the picrorhizon extract significantly reduces the skin oil content at 4h and 8h, specifically by 21.41% and 21.49%, respectively. The photos of the test formula containing the picrorhizon extract at 0 days and 28 days show that the oiliness of the skin is significantly reduced after using the test sample for 28 days. This shows that the picrorhizon extract has obvious oil control effect and can be used as an effective ingredient in daily chemical products.

[0063] In the present invention, the oil-controlling mechanism of the picrorhizon extract is analyzed by transcriptomics, showing that the picrorhizon extract affects lipid synthesis by regulating the PPARγ signaling pathway, and ultimately reduces the oil secretion of the skin, providing a scientific basis at the genetic level for the application of the picrorhizon extract in oil-controlling daily chemical products.

[0064] In the present invention, the picrorhizoma berberis extract has a significant inhibitory effect on ROS, especially the picrorhizoma berberis extract with a concentration of 0.4-0.6 mg / mL has a more significant inhibitory effect on ROS, indicating that the picrorhizoma berberis extract has anti-inflammatory, soothing, radiation protection and anti-aging functions on the skin.

[0065] Picroside I and Picroside II

[0066] From a structural analysis, picroside I has an additional set of double bonds and reduces the phenolic hydroxyl group on the benzene ring. Theoretically, its polarity is smaller than that of picroside II. Combined with the peak order of the HPLC detection spectrum, it can be basically inferred that the use of low concentration ethanol makes it easier to elute picroside II, which is consistent with the results of comparative experiment 3.

[0067] In addition, because picroside I has an extra set of double bonds, the conjugated system formed with the benzene ring and ketone carbonyl group is more sensitive to ultraviolet absorption. Actual test results also confirm that for the same concentration of picroside I, the peak area in the HPLC test spectrum is almost twice that of picroside II.

[0068] Compared with the prior art, the present invention has the following beneficial effects: 1. In the present invention, the oil control effect of the picrorhizon extract was evaluated through human efficacy tests. The results showed that the picrorhizon extract could significantly reduce skin oil and skin oiliness, indicating that the picrorhizon extract has obvious oil control effect and can be used as an effective ingredient in daily chemical products.

[0069] 2. In the present invention, the oil-control mechanism of the picrorhizon extract was analyzed through transcriptomics, showing that the picrorhizon extract can significantly inhibit the expression of the fatty acid binding protein FABP4 gene in the lipid metabolism PPAR signaling pathway, ultimately reducing the oil secretion of the skin, providing a scientific basis at the genetic level for the application of the picrorhizon extract in oil-control daily chemical products.

[0070] 3. In the present invention, through transcriptomic analysis, it was also found that the picrorhizon extract can down-regulate the expression of genes FOS, TNF, and IL21R related to inflammation, and can also down-regulate matrix metalloproteins MMP1 and MMP10 related to aging, indicating that the picrorhizon extract has anti-inflammatory and anti-aging effects at the genetic level.

[0071] 4. In the present invention, the efficacy of the picrorhizon extract in reducing the level of active oxygen and reducing the damage of ultraviolet rays to cells is evaluated, indicating that the picrorhizon extract has anti-inflammatory, soothing, radiation protection and anti-aging functions on the skin.

[0072] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually based on conventional conditions or the conditions recommended by the manufacturer. Unless otherwise stated, percentages and parts are by weight.

[0073] Comparative experiment 1: Extraction solvent and its concentration screening In the early stage, the content of picroside I in the medicinal material was determined to be 3.39%, and the content of picroside II was 8.25%. Take 30g of picroside, use water, 50% and 70% ethanol as extraction solvents, respectively, and heat under normal pressure for reflux extraction twice. The solid-liquid ratio of the first reflux extraction is 1:10, the heating temperature is 85℃, and the extraction time is 2h; the solid-liquid ratio of the second reflux extraction is 1:8, the heating temperature is 85℃, and the heating time is 1h; filter with filter cloth to obtain a crude extract. The crude extract is concentrated under reduced pressure, the concentration temperature is 55-60℃, the vacuum degree is -0.08MPa, and it is concentrated to twice the amount of feed to obtain a crude extract. Weigh and determine the content of active ingredients by HPLC. The specific data are shown in the following table.

[0074] Table 1 Yield, active ingredient content and transfer rate at the tested concentrations

[0075] From the data in the table, it can be seen that with the increase of ethanol concentration, the transfer rate of picroside I and picroside II is significantly improved. 70% ethanol as the extraction solvent is better than low-concentration ethanol such as 50% ethanol and water in extracting the target components.

[0076] Comparative experiment 2: Static adsorption experiment of macroporous resin The resin static adsorption method was used to investigate the adsorption and desorption abilities of D101, AB-8, HP20, HPD100 and DM130 for picroside I and picroside II.

[0077] Take the lyophilized powder of crude extract of Coptis chinensis (prepared from Coptis chinensis medicinal materials with 4.08% picroside I and 10.13% picroside II, and obtain lyophilized powder with 7.94% picroside I and 18.79% picroside II) as raw material, weigh 1.5g, add 5mL of water to dissolve completely, and prepare 6 portions. Mix with 10mL macroporous resin respectively, place on a shaker (speed 160rpm) for static adsorption for 30min, and filter respectively to obtain each filtrate. Take the resin after static adsorption, add water to analyze three times, 10mL each time, wash with water three times, add 70% ethanol to analyze three times, 10ml each time, combine each water part, dilute to 100ml for detection, combine each 70% ethanol, dilute to 100ml for detection.

[0078] Adsorption rate = (1-mass of target component in water part / total mass of target component) × 100% Resolution = (70% target component mass / total target component mass) × 100% Table 2 Adsorption and analysis of active ingredients by each resin under static adsorption

[0079] From the data in the table, when the resin models are D101, AB-8, and HP20, the adsorption rates of picroside I and picroside II are significantly higher than those of other resins, and the adsorption rates are all greater than 90%. Among them, the HP20 resin has a slightly higher adsorption rate for active ingredients, especially picroside II, than D101 or AB-8, and the resolution rate is also high, and the active ingredients are easy to elute. Therefore, HP20 was selected for the next step of the experiment.

[0080] Comparative experiment 3: Screening of solvents for elution of macroporous resin Weigh the extract of Picrorhizoma Coptidis (prepared from Picrorhizoma Coptidis with 4.08% picroside I and 10.13% picroside II, and obtain 18.38g of dry matter, of which picroside I is 8.21% and picroside II is 19.42%) and load it into 200mL of D101 macroporous resin. After loading, elute with water, 30% ethanol, and 50% ethanol in turn, eluting 3BV each time. Collect each elution part, concentrate, dry, weigh, and test. The experimental results are as follows:

[0081] Table 3 Results of elution solvent screening experiment

[0082] From the data in the table, it can be seen that the elution efficiency of 30% ethanol is relatively low, especially the elution ability of picroside I is relatively weak. In the prior art, low-concentration ethanol is used for elution, but the inventor unexpectedly found that the elution efficiency of low-concentration ethanol in the method of the present invention is not as good as that of high-concentration ethanol, especially the elution efficiency of the more effective component picroside I is low, so high-concentration (70%) ethanol is used as the extraction and elution solvent in the final solution.

[0083] Example 1 Preparation of Picrorhizoma Extract In this experiment, picrorhizon extract was prepared by extracting picrorhizon slices. The extraction steps include:

[0084] (1) Extraction: Mix picrorhizoma berberis tablets (from artificial cultivation in Yunnan) with 70% ethanol solution, and perform reflux extraction twice at normal pressure. The solid-liquid ratio of the first reflux extraction was 1:10, the heating temperature was 85°C, and the extraction time was 2 h; the solid-liquid ratio of the second reflux extraction was 1:8, the heating temperature was 85°C, and the heating time was 1 h; filter with filter cloth to obtain a crude extract.

[0085] (2) Concentration: The crude extract is concentrated under reduced pressure at a temperature of 55-60°C and a vacuum degree of -0.08 MPa, and concentrated to twice the amount of feed to obtain a crude extract concentrate.

[0086] (3) Resin purification: The crude extract was loaded onto a HP20 macroporous resin and first eluted with water at a volume of 3 BV; then eluted with 70% ethanol at a volume of 3 BV. The ethanol eluate was collected and concentrated under reduced pressure to obtain the extract of Picrorhizoma Coptidis.

[0087] (4) Freeze drying: The extract of the coptis chinensis extract is freeze dried in a vacuum freeze dryer for more than 24 hours. The freezing temperature of the freeze drying is (-70°C to -50°C) and the vacuum degree is less than 20 Pa to obtain the lyophilized powder of the coptis chinensis extract.

[0088] The prepared lyophilized powder of Picrorhizoma Coptidis extract was tested by HPLC. The results were as follows: Figure 1 The results showed that the content of picroside I was 15.68wt%, the content of picroside II was 34.67wt%, and the ratio of picroside I to picroside II in the Picrorhizoma Coptidis extract obtained in this experiment was 3:7.

[0089] Example 2 Oil Control Human Efficacy Test Test product formula: 1,3-butylene glycol 3%, water 95.9%, tamarind gum 0.1%, 1,2-hexanediol 0.8%, p-hydroxyacetophenone 0.3%, picrorhizon extract 0.1%.

[0090] Blank control formula: 1,3-butanediol 3%, water 96%, tamarind gum 0.1%, 1,2-hexanediol 0.8%, and p-hydroxyacetophenone 0.3%.

[0091] Subjects: 15 volunteers with oily skin aged between 24 and 30 were screened.

[0092] Test equipment: CK skin microscope Courage + Khazaka electronic Visioscope PC 35 Test method: 2.1 Short-term oil control experiment Clean the test area of ​​the test subject, rinse it with clean water and dry it with a lint-free absorbent paper towel. After 30 minutes, measure the amount of oil on the skin surface of the sample application area and the control area respectively. Measure 3 times at different positions in each area. The measurement result is expressed as the average value of 3 measurements as the initial value.

[0093] Add 0.5 mL of the test formula to a clean cotton pad, and add 0.5 mL of the blank control formula to another clean cotton pad. Apply the test formula containing the picrorhizome extract to area A, and apply the blank control to area B (such as Figure 2 The skin was placed in a constant temperature and humidity room (temperature 21±1℃, humidity 50±5%) for 10 min and then removed. The skin oil production was tested 4h and 8h later on the same day.

[0094] 2.2 Long-term oil control experiment After cleansing with water at night, the subjects evenly applied essence on their face once. The amount of experimental sample used each time was 1.5±0.5mL, and the application time was 10min each time. During the experiment, no other medicines or cosmetics that may affect the experimental results were used. After 28 consecutive days of use, the amount of oil on the subjects' skin surface was tested according to the short-acting oil control test method. The amount of oil on the skin surface of the blank group tested at 4h and 8h was compared and analyzed with the test data of the blank group at 4h and 8h on day 0.

[0095] 2.3 Short-term oil control test results: (1) Statistics of skin oil secretion of 15 subjects 4h and 8h after using the test product formula Table 4 Statistics of skin oil secretion after 4h and 8h using the test product formula

[0096] (2) Statistical analysis of short-term oil control experimental data like Figure 3As shown, compared with the blank control formula, the test formula containing picrorhizoma berberis extract significantly reduced the skin oil content at 4h and 8h, with average values ​​of -21.41% and -21.49% respectively. This shows that the test formula containing picrorhizoma berberis extract has a good oil control effect.

[0097] 2.4 Long-term oil control test results: (1) If Figure 4 As shown, compared with the test data of the blank group at 4h and 8h on day 0, the comparative analysis of the amount of oil on the skin surface tested at 4h and 8h in the test group using the test sample for 28 days showed that the oil production of the skin of the test group decreased, indicating that the test formula containing Picrorhizoma Ceratitis extract has a good oil control effect.

[0098] (2) Volunteer test photos like Figure 5 As shown, by comparing the photos of the test subjects using the test formula containing Picrorhizoma Coptidis extract for 0 days and 28 days, it was found that after using the test sample for 28 days, the oiliness of the skin was significantly reduced.

[0099] Example 3 Cellular transcriptome test of Picrorhizoma Coptidis extract Test model: UVB-induced keratinocyte photoaging model Test data analysis: Figure 6 As shown, compared with the blank control group (NC group), in the transcriptomic KEGG enrichment of 0.5 mg / mL lyophilized powder of Picrorhizoma Coptidis extract, genes related to immunity, endocrine, fat metabolism, signal transduction, cell growth and death were significantly enriched.

[0100] Further analysis of gene expression data revealed that, compared with the blank control group, treatment with 0.5 mg / mL of lyophilized powder of Picrorhizoma Coptidis extract significantly inhibited the gene expression of fatty acid binding protein FABP4 in the lipid metabolism PPAR signaling pathway ( Figure 7 A) By analyzing the transcriptomic data, after adding picrorhizome extract, the expression of PPARGC1A gene in the PPARγ pathway was significantly inhibited, and the expression of PPARGC1A gene was not detected in the experiment ( Figure 7 B). Therefore, it is speculated that the extract of Picrorhizoma Coptidis may inhibit the synthesis and secretion of lipids by inhibiting the PPARγ signaling pathway.

[0101] Compared with the blank control group, the inflammation-related genes FOS, TNF, and IL21R in cells were significantly downregulated after treatment with 0.5 mg / mL lyophilized powder of Picrorhizoma Coptidis extract ( Figure 8 ).

[0102] In addition, compared with the blank control group, after treatment with 0.5 mg / mL of lyophilized powder of Picrorhizoma Coptidis extract, the aging-related matrix metalloproteins MMP1 and MMP10 in the cells showed a significant downregulation trend ( Fig. 9 ).

[0103] Example 4 Experiment on reducing ROS secretion in cells induced by UVB irradiation Sample group: The lyophilized powder of Picrorhizoma Coptidis extract was prepared with basal culture medium to prepare test samples of different concentrations.

[0104] Blank control group: select basal culture medium.

[0105] Positive control group: Vitamin E (PC) was prepared into 200 μg / ml using basal culture medium.

[0106] 4.1 Main reagents and instruments MEM culture medium, fetal bovine serum, MTT, PBS (PH7.4), vitamin E, pipette, microplate reader, inverted microscope, clean bench, ultrapure water machine, CO2 constant temperature incubator, ultra-low temperature freezing storage box, etc.

[0107] 4.2. Determination of maximum safe concentration by MTT method Human keratinocytes (Hacat) growing in logarithmic phase were cultured at 8 × 10 4 The cells were inoculated at a density of 1.5 cell / ml in a 96-well plate, and a blank control group was set up. After culturing for 24 h at 37°C and 5% CO2, different concentrations of the test substances were added to the cells as the sample group, and the culture medium was used to replace the sample as the negative control group. After culturing for 24 h, 20 μL of MTT solution was added to each well, and the culture was continued for 4 h. The liquid in the wells was discarded, and 150 μL of DMSO solution was added to each well. The plates were oscillated at room temperature for 10 min, and the absorbance was measured at 490 nm.

[0108] According to: Cell viability % = (OD value of sample group - OD value of blank group) / (OD value of negative control group - OD value of blank group) × 100%. Calculate the cell viability at each concentration.

[0109] 4.3 Test steps Hacat cells in the logarithmic growth phase were inoculated into well plates and cultured at 37°C and 5% CO2 for 24 h. Then, different concentrations of the test substances were added, and negative and positive control groups were set up. After 1 h, the test substances were irradiated with 50 mJ / cm 2 After being stimulated by UVB, the cells were returned to the incubator for 24 h and the fluorescence intensity of the cells in each test group was detected.

[0110] 4.4 Experimental Results 4.4.1 MTT assay to determine cell viability, such as Fig.10 As shown in Table 5, sample concentrations ≤ 0.5 mg / ml had no significant effect on the viability of Hacat cells.

[0111] Table 5 Effects of different samples on the viability of Hacat cells

[0112] 4.4.2 DCFH-DA fluorescence method was used to determine the inhibitory effect of lyophilized powder of Picrorhizon extract on ROS secretion. Fig.11 As shown, compared with the blank control group (BC group), the secretion of ROS after UVB stimulation (NC group) increased significantly (###P<0.001), and the PC group could significantly inhibit the expression of ROS (###P<0.001), indicating that the stimulation conditions were effective. The 0.5 mg / mL concentration of the picrorhizome extract had a significant inhibitory effect on the ROS secreted by Hacat cells induced by UVB, with an inhibition rate of 23.20%. This shows that the picrorhizome extract of the present invention has the function of reducing the level of active oxygen and reducing the damage of ultraviolet rays to cells.

[0113] Example 5 Experiment on reducing the secretion of inflammatory factors TNF-α and IL-6 Sample group: The lyophilized powder of Picrorhizoma Coptidis extract was prepared with basal culture medium to prepare test samples of different concentrations.

[0114] Blank control group: select basal culture medium.

[0115] Positive control group: Dexamethasone (PC) was prepared at 100 μg / ml in basal medium.

[0116] 5.1 Main reagents and instruments DMEM high-glucose culture medium, fetal bovine serum, MTT, PBS, dexamethasone, pipette, microplate reader, inverted microscope, clean bench, ultrapure water machine, CO2 constant temperature incubator, ultra-low temperature freezing storage box, etc.

[0117] 5.2 Test steps Raw264.7 cells in the logarithmic growth phase were cultured in a plate at 37°C and 5% CO2 for 24 hours, and then different concentrations of the test substances were added. A negative control group and a positive control group were set up. LPS stimulation was added 1 hour later. The cells were returned to the incubator and incubated for 24 hours. The cell supernatants corresponding to each sample group were collected, and the concentrations of TNF-α and IL-6 in the cell supernatants were determined using ELISA kits.

[0118] 5.3 Experimental Results The results of cell viability experiments were as follows Fig.10 As shown, sample concentration ≤ 50 μg / ml had no significant effect on cell viability.

[0119] The results of the inflammatory factor secretion experiment are as follows Fig.12 , 13 As shown, compared with the blank control group (BC group), the secretion of TNF-α and IL-6 increased significantly after LPS stimulation (NC group) (###P< 0.001), indicating that the stimulation conditions were effective. The IL-6 concentration in the positive control group (PC group) and the experimental group was significantly lower than that in the NC group. The positive control dexamethasone (100μg / ml) significantly inhibited the secretion of TNF-α and IL-6 (***P< 0.001). 50μg / ml of lyophilized powder of Picrorhizoma Coptidis extract can significantly inhibit / reduce the secretion of TNF-α and IL-6, of which the TNF-α secretion inhibition rate was 28.59% and the IL-6 secretion inhibition rate was 44.50%. It can be seen that the Picrorhizoma Coptidis extract of the present invention has anti-inflammatory activity.

[0120] discuss In the present invention, a new function and use of a picrorhizon extract is provided, specifically involving the application of the picrorhizon extract as an oil-controlling substance in daily chemical products; the picrorhizon extract reduces the secretion of inflammatory factors TNF-α and IL-6, thereby playing an anti-inflammatory and soothing role; the picrorhizon extract reduces the level of active oxygen and reduces the damage of ultraviolet rays to cells, thereby playing a radiation protection and anti-aging function for the skin.

[0121] Wild picrorhizon is a national second-class protected species, native to Tibet and northwest Yunnan at an altitude of 3900-4500m, with a narrow distribution area. Due to long-term over-exploitation, wild resources are on the verge of extinction. At present, picrorhizon has been cultivated on a large scale in Yunnan. The present invention expands the application of picrorhizon extract in daily chemical products, provides new ideas for the application of daily chemical products for oil control, anti-inflammatory and soothing, radiation protection, and anti-aging, promotes the healthy development of picrorhizon cultivation industry, and realizes the protection and sustainable utilization of endangered medicinal plant resources.

[0122] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A picrophora extract, characterized in that The picroside extract contains picroside I and picroside II, the content of picroside I and picroside II in the extract is 40%-60%, and the mass ratio of picroside I to picroside II is 3-4:7; The picrorhizoma berberis extract is prepared by the following method: (1) Extraction: providing a mixture of Picrorhizoma Coptidis raw materials and ethanol, and performing reflux extraction for 1-3 times to obtain a crude extract; (2) Concentration: Concentrating the crude extract obtained in step (1) under reduced pressure to obtain a crude extract concentrate; (3) Resin purification: The crude extract obtained in step (2) is loaded onto a macroporous resin, first eluted with water and then with ethanol, the ethanol eluate is collected, and concentrated under reduced pressure to obtain a Picrorhizoma Coptidis extract; (4) freeze-drying: freeze-drying the picrorhizoma picrorhizoma extract obtained in step (3) to prepare a picrorhizoma picrorhizoma extract freeze-dried powder, i.e., the picrorhizoma picrorhizoma extract; Wherein, the concentration of ethanol in step (1) is 65 v / v%-75 v / v%; The macroporous resin in step (3) is selected from the group consisting of HP20, AB-8, and D101 macroporous resins; The ethanol concentration of the ethanol elution in step (3) is 65 v / v%-75% v / v ethanol.

2. The Picrorhizoma Coptidis extract according to claim 1, characterized in that The content of picroside I and picroside II is 45%-55%, based on the total mass of the picroside extract.

3. The method for preparing the Picrorhizoma Coptidis extract according to claim 1, characterized in that: The method comprises the steps of: (1) Extraction: providing a mixture of Picrorhizoma Coptidis raw materials and ethanol, and performing reflux extraction for 1-3 times to obtain a crude extract; (2) Concentration: Concentrating the crude extract obtained in step (1) under reduced pressure to obtain a crude extract concentrate; (3) Resin purification: The crude extract obtained in step (2) is loaded onto a macroporous resin, first eluted with water and then with ethanol, the ethanol eluate is collected, and concentrated under reduced pressure to obtain a Picrorhizoma Coptidis extract; (4) Freeze-drying: freeze-drying the picrorhizoma picrorhizoma extract obtained in step (3) to prepare lyophilized powder of picrorhizoma picrorhizoma extract, i.e., the picrorhizoma picrorhizoma extract.

4. The method according to claim 3, characterized in that The reflux extraction in step (1) is performed twice, the solid-liquid ratio of the first reflux extraction is 1:9-11, the heating temperature is 80-90°C, and the extraction time is 1.5-2.5h; the solid-liquid ratio of the second reflux extraction is 1:7-9, the heating temperature is 80-90°C, and the heating time is 0.5-1.5h; filtering is performed using filter cloth to obtain a crude extract.

5. The method according to claim 3, characterized in that The macroporous resin in step (3) is HP20.

6. The use of the Picrorhizoma Coptidis extract according to claim 1, characterized in that: Used for preparing a composition for lowering skin oil content.

7. The use according to claim 6, characterized in that The composition is also used to modulate the PPARγ signaling pathway, thereby affecting lipid synthesis.

8. The use according to claim 6, characterized in that The compositions are also useful for reducing active oxygen levels.

9. A composition, characterized in that The composition comprises: (1) the Picrorhizoma Coptidis extract according to claim 1 or 2; and (2) additional ingredients that can be used in daily chemical products.

10. The composition according to claim 9, characterized in that The composition is used for purposes selected from the group consisting of oil control, anti-inflammatory and soothing, radiation protection, anti-aging, or a combination thereof.

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