CN112972524B - Application of schisandra chinensis oil in inhibition of sebaceous gland activity - Google Patents
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Abstract
The application belongs to the technical field of biological medicines. The application provides application of schisandra chinensis oil in inhibiting sebaceous gland activity. Experimental results show that the schisandra chinensis oil can obviously inhibit the proliferation of sebaceous gland cells, inhibit the lipid synthesis of the sebaceous gland cells and reduce the expression of sebaceous gland cell receptors at the cellular level, and realizes the effects of inhibiting the growth of sebaceous gland plaques and inhibiting the sebum secretion, so that the schisandra chinensis oil has the effects of inhibiting the sebaceous gland activity, controlling oil, regulating the water-oil balance of the face and the like, can be applied to medicaments for treating acne or oil-controlling cosmetics, and has the advantages of safety, high efficiency and low cost.
Description
Technical Field
The application belongs to the technical field of biomedicine, and particularly relates to application of schisandra chinensis oil in inhibition of sebaceous gland activity.
Background
Schisandra chinensis (Schisandra chinensis) is a dry mature fruit of Schisandra chinensis (Schisandra chinensis) belonging to Magnoliaceae, and was listed as the first-grade product in Shen nong Ben Cao Jing. The book Xin Xiu Ben Cao records that the five flavors of skin and meat are sweet and sour, and bitter and pungent in kernel, all have salty taste, so it is named as Wu Wei Zi. The schisandra fruit has the functions of nourishing and strengthening and has high medicinal and edible values. Chemical research shows that the schisandra chinensis mainly contains lignans, volatile oils, flavonoids, terpenes, organic acids and other active ingredients, and pharmacological research shows that the schisandra chinensis has the effects of protecting liver, tranquilizing and hypnotizing, reducing blood sugar, resisting oxidation, enhancing immunity, resisting cancer, resisting AIDS virus and the like. However, the schisandra chinensis has not been reported to inhibit the sebaceous gland activity.
The main components of sebum include triglyceride, wax ester, squalene, cholesterol ester and cholesterol. Sebaceous glands are a kind of glandular vesicle-like full-plasma secretory tissue distributed in the skin of the whole body except palms and soles, and have the highest density on the scalp and face. The normal lipid on the surface of the skin can play a barrier role, can moisten the skin, resist infection and maintain the surface bacterial state balance of the human skin. Excessive sebum secretion from sebaceous glands leads to excessive skin oil production and aesthetic problems, and may also promote the development of acne or other diseases, which seriously affect the mental health and quality of life of patients.
The breakdown of keratinocytes and the sebum secreted by sebaceous glands constitute the main source of lipids at the skin surface. Among them, the breakdown of keratinocytes produces only a small fraction of the lipids, and the excessive sebum secretion from the sebaceous glands is the main cause of the greasiness of the skin. The sebaceous gland function is regulated by a plurality of factors, and androgen is taken as a primary factor influencing sebum secretion and mainly regulates the differentiation and proliferation of the sebaceous gland and the synthesis and secretion of sebum. In addition, sebaceous gland function is also regulated by enzyme systems such as Peroxisome Proliferator Activated Receptors (PPARs), neuropeptides, antimicrobial peptides, inflammatory cytokines, and the like.
At present, western medicines are mainly used for effectively inhibiting sebaceous gland secretion, and have the defects of multiple side effects and strong dependence, so that a safe and effective substitute component is urgently needed.
Disclosure of Invention
In view of the above, the present application provides an application of schizandra chinensis oil in inhibiting sebaceous gland activity, wherein the schizandra chinensis oil can significantly inhibit growth of sebaceous gland plaques and sebum secretion at an animal level.
The specific technical scheme of the application is as follows:
in a first aspect, the present application provides the use of schizandra chinensis oil for inhibiting sebaceous gland activity.
Preferably, the inhibition of sebaceous gland activity is specifically inhibition of sebaceous gland cell proliferation.
Preferably, the inhibition of sebaceous gland activity is specifically inhibition of sebaceous gland cell lipid synthesis.
Preferably, the inhibition of sebaceous gland activity is specifically the down-regulation of the expression of androgen receptor mRNA of sebaceous gland cells.
Preferably, the schisandra chinensis oil is volatile oil obtained by extracting schisandra chinensis fruits.
Preferably, the step of extracting comprises: crushing the schisandra chinensis, sieving the crushed schisandra chinensis with a 14-60-mesh sieve, adding the crushed schisandra chinensis into an aqueous solution of NaCl according to the liquid-material mass ratio of 4-8 times, soaking for 2-4 h, then carrying out water distillation for 2-5 h, and collecting the upper essential oil to obtain the schisandra chinensis.
More preferably, the step of extracting comprises: pulverizing fructus Schisandrae, sieving with 40 mesh sieve, adding into 6 times of distilled water containing 6% NaCl, soaking for 3 hr, distilling with water for 3 hr, collecting upper layer essential oil, and adding anhydrous NaSO 4 Drying and filtering to obtain the product.
In the application, animal level experiments show that the schisandra chinensis oil can obviously inhibit the proliferation of sebaceous gland cells, inhibit the lipid synthesis of the sebaceous gland cells and reduce the expression of a sebaceous gland cell receptor at the cellular level, so that the effects of inhibiting the growth of sebaceous gland plaques and inhibiting the secretion of sebum are realized, and the schisandra chinensis oil has good development potential.
In a second aspect, the present application provides the use of schizandra chinensis oil for the manufacture of a medicament for the treatment of acne.
Preferably, the dosage form of the medicament is emulsion or paste.
In a third aspect, the application provides application of schisandra chinensis oil in preparing oil-control cosmetics.
Preferably, the cosmetic is in the form of water, lotion, essence or facial mask.
In the application, the schisandra chinensis oil can obviously inhibit the growth of sebaceous gland plaques and sebum secretion at the animal level, so that the schisandra chinensis oil has the effects of inhibiting the activity of sebaceous glands, controlling oil, regulating the water-oil balance of the face and the like, can be applied to medicaments for treating acne or oil-control cosmetics, and has the advantages of safety, high efficiency and low cost.
In summary, the application provides an application of schisandra chinensis oil in inhibition of sebaceous gland activity. Experimental results show that the schisandra chinensis oil can obviously inhibit the proliferation of sebaceous gland cells, inhibit the lipid synthesis of the sebaceous gland cells and reduce the expression of sebaceous gland cell receptors at the cellular level, and realizes the effects of inhibiting the growth of sebaceous gland plaques and inhibiting the sebum secretion, so that the schisandra chinensis oil has the effects of inhibiting the sebaceous gland activity, controlling oil, regulating the water-oil balance of the face and the like, can be applied to medicaments for treating acne or oil-controlling cosmetics, and has the advantages of safety, high efficiency and low cost.
Drawings
In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings needed to be used in the description of the embodiments or the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present application, and it is obvious for those skilled in the art that other drawings can be obtained according to the drawings without inventive exercise.
FIG. 1 is the sebum secretion amount (left side) of sebaceous plaques of golden hamster at 0, 10, 20, 30d before and after administration of Schisandra chinensis oil in example 5;
FIG. 2 is the sebum secretion amount of sebaceous gland plaques of mice with golden yellow color at 0, 10, 20, 30d before and after administration of the schisandra fruit oil in example 5 (right side).
Detailed Description
In order to make the objects, features and advantages of the present application more obvious and understandable, the technical solutions in the embodiments of the present application are clearly and completely described, and it is obvious that the embodiments described below are only a part of the embodiments of the present application, and not all embodiments. All other embodiments obtained by a person of ordinary skill in the art based on the embodiments in the present application without making any creative effort belong to the protection scope of the present application.
The starting materials and reagents used in the examples of the present application are commercially available, and the experimental procedures used are routine in the art unless otherwise specified.
The golden hamster of the embodiment of the application is purchased from Guangdong province medical animal experiment center, SZ95 cells are purchased from Shenzhen Zhonghongbo Biotechnology Co., ltd, propylene glycol is purchased from Tianjin Mao chemical reagent factory, chloral hydrate is purchased from Sigma-Aldrich, DMSO is purchased from Tianjin Mao chemical reagent factory, nile Red fluorescent dye solution is purchased from Shanghai Haoling Biotech Co., ltd, reverse transcription kit is purchased from Promega Co., trizol is purchased from Gibco Co., dNTP is purchased from Guangdong Sheng Biotechnology Co., mgCl 2 Purchased from Shanghai Aladdin Biotechnology Ltd.
Example 1 extraction of Schisandra chinensis oil
Pulverizing fructus Schisandrae chinensis, sieving with 40 mesh sieve, adding into 6 times of liquid-material ratio, and distilling with 6% NaClSoaking in water for 3 hr, distilling with water for 3 hr, collecting upper layer essential oil, and treating with anhydrous NaSO 4 Drying and filtering to obtain the product.
Example 2 Effect of Schisandra chinensis oil on the proliferation of sebaceous gland cells
(1) Culturing human sebaceous gland cells (SZ 95 cells), and digesting the SZ95 cells by trypsin to prepare SZ95 cell suspension when the cells grow and fuse to 80%;
(2) Inoculating the SZ95 cell suspension to a 96-well culture plate, wherein 4000 cells are cultured in each well, removing the supernatant after 48 hours of culture, washing the supernatant for 2 times by using a PBS solution, respectively adding culture media containing 0.5, 5, 50 and 500 mu g/mL of the schisandra chinensis oil prepared in the example 1, continuing to culture for 48 hours, removing the supernatant, and washing the supernatant for 2 times by using the PBS solution;
(3) Adding 0.5mg/mL MTT 100u1, culturing for 4h, discarding MTT, adding 150 μ LDMS0 per well, and incubating for 10min. The absorbance was measured using a microplate reader at a wavelength of 490nm.
3 parallel wells are set for each action concentration, a medium without the addition of the schizandra oil is used as a blank group, a medium without the addition of SZ95 cells and the addition of the schizandra oil is used as a blank control group, and a medium with each action concentration is used as a zero adjustment group while the addition of the SZ95 cells is not performed. The results of the effect of schizandra chinensis oil on the proliferation rate of SZ95 cells in example 2 of the present application are shown in table 1, wherein the proliferation rate of SZ95 cells in the blank group = blank group OD-blank control group OD, and the proliferation rate of SZ95 cells in the experimental group = (experimental group OD-zero-adjustment group OD)/(blank group OD-blank control group OD).
TABLE 1 Effect of Schisandra chinensis oil on the proliferation Rate of SZ95 cells
| Group of | SZ95 cell proliferation Rate (%) |
| Blank group | 100±4.8860 |
| 0.5. Mu.g/ml test group | 96.67±4.0787 |
| 5 μ g/ml test group | 86.10±4.0398* |
| 50 μ g/ml test group | 71.09±7.6650*** |
| 500 μ g/ml experimental group | 46.71±8.3923*** |
Note: p < 0.05, P < 0.001, compared to blank
As can be seen from Table 1, 0.5. Mu.g/ml schizandra chinensis oil has no significant inhibition effect on the proliferation of sebaceous gland cells (P is more than 0.05); the fructus schizandrae oil with the concentration of 5 mu g/ml has obvious inhibition effect on the proliferation of sebaceous gland cells (P is less than 0.01); 50. the Schisandra chinensis oil with the concentration of 500 mu g/ml has stronger inhibition effect on the proliferation of sebaceous gland cells (P is less than 0.001).
Example 3 Effect of Schisandra chinensis oil on lipid Synthesis in sebaceous gland cells
(1) Culturing SZ95 cells, and digesting the SZ95 cells by trypsin to prepare SZ95 cell suspension when the cells grow and fuse to 80%;
(2) SZ95 cells were seeded in 6-well plates at 2X 10 per well 5 After further culturing for 48 hours, the medium containing 0.5, 5, 50 and 500. Mu.g/mL of the Schisandra chinensis oil prepared in example 1 was added. Continuing culturing for 48h, adding trypsin for digestion, stopping digestion with a culture medium containing 10% fetal calf serum, and washing with PBS solution for 2 times to prepare single cell suspension;
(3) Nile red fluorescent dye solution was added to a final concentration of 100ng/mL. Incubate at room temperature for 15min, and filter with 300 mesh nylon filter. The fluorescence intensity of 10000 cells per group was measured by flow cytometry. Finally, the average fluorescence intensity (excitation wavelength 485nm, emission wavelength 565 nm) of each cell was calculated.
3 parallel wells were set for each effect concentration, and medium without schizandra oil was added as a blank. The results of the effect of schizandra chinensis oil on the synthesis of SZ95 cell lipids in example 3 of the present application are shown in table 2, wherein the inhibition rate of the experimental group = (1-experimental group fluorescence unit/blank group fluorescence unit) × 100%.
TABLE 2 Effect of Schisandra chinensis oil on SZ95 cell lipid Synthesis
| Group of | Mean fluorescence unit | Inhibition ratio (%) |
| Blank group | 649.33±46.0145 | - |
| 0.5. Mu.g/ml test group | 629.67±28.1484 | 3.03 |
| 5 μ g/ml test group | 574.90±32.4553 | 11.46 |
| 50 μ g/ml experimental group | 458.21±52.0152*** | 29.43 |
| 500 μ g/ml experimental group | 497.50±44.9246** | 23.38 |
Note: p < 0.01, P < 0.001, compared to blank
As can be seen from Table 2, 0.5 and 5. Mu.g/ml of schizandra chinensis oil had insignificant inhibitory effects (P > 0.05) on sebaceous gland cell lipid synthesis; the inhibitory effect of 50 and 500 mu g/ml schisandra fruit oil on the proliferation of sebaceous gland cells is obvious (P < 0.01 and P < 0.001 respectively).
Example 4 Effect of Schisandra chinensis oil on the expression of Androgen Receptor (AR) mRNA of sebaceous gland cells
(1) Culturing SZ95 cells, and digesting the SZ95 cells by trypsin to prepare SZ95 cell suspension when the cells grow and fuse to 80%;
(2) The SZ95 cells were seeded in 6-well plates at 2X 10 cells per well 5 After culturing for 96 hours, the medium containing 0.5, 5, 50 and 500. Mu.g/mL of the schizandra chinensis oil prepared in example 1 was added. After further incubation for 24h, trizol was added and total RNA was extracted from SZ95 cells according to the instructions of the reagents. cDNA was synthesized by reverse transcription following the reverse transcription kit instructions, followed by in-tube amplification of AR and β -actin (β -actin). The primer sequence is as follows: the AR sense strand: 5 'GAAGACCTGCCTGATCTGTG-3', antisense strand: 5 '-AAGCCTCTCTCCITCCTCTGT-3' (product length 269 bp); internal reference beta-actin sense strand: 5 'AGAGAGATGGCCACGGCTGCTT-3', antisense strand: 5 'ATTTGCGGTGGACGATGGAG-3' (product length 445 bp). The reaction system is 25 μ L: wherein PCR 10 Xbuffer 2.5. Mu.L, 10mmol/L dNTP mixture 0.5. Mu.L, 25mol/L MgCl 2 mu.L of AR sense and antisense strands (20 pmoL/L) each 1.2. Mu.L, beta-actin sense and antisense strands (20 pmoL/L) each 0.2. Mu.L, taq DNA polymerase 1.25U, cDNA 10. Mu.L, and finally deionized water to a total volume of 25. Mu.L. The reaction conditions are as follows: 30s at 94 ℃ 30s, 45s at 60 ℃ and 30s at 72 ℃ for a total of 40 cycles. The RT-PCR products were electrophoresed on a 20% agarose gel and stained with 0.5% ethidium bromide. And analyzing the scanning result by adopting a Shanghai energy gel imaging analysis system.
3 parallel wells were set for each effect concentration, and medium without schizandra oil was added as a blank. The absorbance value takes the product of the intensity and the net area as a quantitative standard, and the ratio of the absorbance value of the specific product and the absorbance value of the beta-actin as a semi-quantitative value. The results of the effect of schizandra chinensis oil on the expression of the androgen receptor mRNA of SZ95 cells in example 4 of the present application are shown in table 3, wherein the expression = experimental group (blank group) absorbance value/β -actin absorbance value.
TABLE 3 Effect of Schisandra chinensis oil on the expression of SZ95 cell Androgen Receptor (AR) mRNA
| Group of | Expression level |
| Blank group | 0.632±0.0215 |
| 0.5. Mu.g/ml test group | 0.642±0.0210 |
| 5 μ g/ml test group | 0.561±0.0331* |
| 50 μ g/ml experimental group | 0.464±0.0319**** |
| 500 μ g/ml experimental group | 0.532±0.0251** |
Note: comparison with blank group, P < 0.05, P < 0.01, P < 0.0001
As can be seen from Table 3, 0.5. Mu.g/ml schizandra oil failed to inhibit the expression of the mRNA of androgen receptor of sebaceous gland cells (P > 0.05); 5. the inhibitory effect of 50 and 500 mu g/ml schisandra fruit oil on the mRNA expression of androgen receptor of sebaceous gland cells is obvious (P is less than 0.05, P is less than 0.0001 and P is less than 0.01 respectively).
Example 5 inhibitory Effect of Schisandra chinensis oil on sebum secretion
(1) Respectively dissolving 25mg and 100mg of the schisandra chinensis oil prepared in the example 1 in propylene glycol, wherein the total volume of the solution is 10mL, and preparing 2.5mg/mL and 10mg/mL schisandra chinensis oil solutions;
(2) 18 male golden yellow hamster are selected, aged for 6 weeks, and the weight of the male golden yellow hamster is 100g +/-20 g, and the male golden yellow hamster is adaptively raised for 3 days before the experiment. The golden hamster was divided into three groups, a blank group (control) was not administered, a low concentration test group (L-SC) was coated with 2.5mg/mL of a schisandra oil solution, and a high concentration test group (H-SC) was coated with 10mg/mL of a schisandra oil solution. 24 hours before application on the first day, the lateral flank hair of each group of golden hamster was shaved and depilatory cream was used to remove hair again, fully exposing the sebaceous glands. Spreading oleum Schisandrae chinensis solution with different concentrations on sebaceous glands of golden hamster 20 μ L each side once a day for 30 days;
(3) After anesthetizing golden hamster by intraperitoneal injection of chloral hydrate at 0, 10, 20 and 30d, respectively, the maximum transverse Diameter (DT) and the maximum longitudinal Diameter (DL) of sebaceous gland plaques were measured with a vernier caliper, and the change of each group of plaques pi × DT × DL was calculated. After anesthetizing the golden hamster by intraperitoneal injection of chloral hydrate for 0, 10, 20 and 30 days, the skin oil and fat were measured by a CK skin tester.
The data are expressed as X + -S, and the difference between groups is compared by t-test. In example 5 of the present application, the comparison of sizes of sebaceous glands of mice with golden yellow color before and after administration of schizandra chinensis oil is shown in table 4 (left side), and the comparison of sizes of sebaceous glands of mice with golden yellow color before and after administration of schizandra chinensis oil is shown in table 5 (right side).
TABLE 4 comparison of size of sebaceous gland plaques of golden hamster before and after administration of Schisandra chinensis oil (left side) (unit: mm) 2 )
| Group of | Number of examples | Area before | Area | 30 days after application |
| Blank group | 6 | 36.82±2.7563 | 56.00±1.8890 | |
| Low concentration test group | 6 | 36.25±1.4849 | 48.00±3.2239** | |
| High concentration test group | 6 | 35.85±3.7546 | 46.03±4.9922** |
Note: p < 0.05, P < 0.01, compared to blank
TABLE 5 comparison of size of sebaceous gland plaque of golden hamster before and after administration of Schisandra chinensis oil (right side) (unit: mm) 2 )
| Group of | Number of examples | Area before application | Area of 30 days after administration |
| Blank group | 6 | 36.54±1.0738 | 57.22±4.2068 |
| Low concentration test group | 6 | 36.45±3.3789 | 53.87±7.8266 |
| High concentration test group | 6 | 36.50±4.0583 | 48.41±4.9341* |
Note: p < 0.05, P < 0.01, compared to blank
As can be seen from tables 4 and 5, the areas of the groups before administration were comparable without significant difference (P > 0.05). The sebaceous gland plaques of the golden hamster of the blank group, the low-concentration experimental group and the high-concentration experimental group are increased compared with those of the golden hamster before treatment, but the sebaceous gland plaques of the low-concentration experimental group and the high-concentration experimental group are reduced compared with those of the blank group, and the inhibition effects of the high-concentration schisandra fruit oil on the sebaceous gland plaque increase are remarkably different (P is less than 0.01).
Sebum secretion amounts of sebaceous gland plaques of golden hamster at 0, 10, 20, 30d before and after administration of schizandra chinensis oil in example 5 of the present application are shown in fig. 1 (left side) and fig. 2 (right side). Fig. 1 and 2 show that sebum secretion from sebaceous plaques on both left and right sides of golden yellow hamster is significantly reduced in the low and high concentration experimental group compared to the blank group after 30 days of administration. Wherein, the high-concentration schisandra chinensis oil has better inhibition effect on sebum secretion of golden hamster.
The experimental results show that the schisandra chinensis oil can inhibit the proliferation of sebaceous gland cells, inhibit the synthesis of lipid by the sebaceous gland cells and reduce the expression of androgen receptor mRNA of the sebaceous gland cells, thereby realizing the effects of inhibiting the growth of sebaceous gland plaques and inhibiting the secretion of sebum and having good development potential.
The above embodiments are only used to illustrate the technical solutions of the present application, and not to limit the same; although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions in the embodiments of the present application.
Claims (4)
1. The application of Schisandra chinensis oil in preparing medicine for treating acne;
the schisandra chinensis oil is volatile oil obtained by extracting schisandra chinensis fruits;
the step of extracting comprises: pulverizing fructus Schisandrae chinensis, sieving with 40 mesh sieve, adding into 6% NaCl aqueous solution according to liquid-material mass ratio of 6 times, soaking for 3 hr, distilling with water for 3 hr, and collecting upper layer essential oil.
2. The use of claim 1, wherein the medicament is in the form of a paste.
3. The application of schisandra chinensis oil in preparing oil-controlling cosmetics;
the schisandra chinensis oil is volatile oil extracted from schisandra chinensis fruits;
the step of extracting comprises: pulverizing fructus Schisandrae chinensis, sieving with 40 mesh sieve, adding into 6% NaCl aqueous solution according to the liquid-material mass ratio of 6 times, soaking for 3h, distilling with water for 3h, and collecting upper layer essential oil.
4. Use according to claim 3, characterized in that the cosmetic formulation is water or an emulsion.
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| FR2948565B1 (en) * | 2009-07-30 | 2011-10-28 | Expanscience Lab | COSMETIC COMPOSITION FOR THE TREATMENT OF ACNE COMPRISING A PETIDIC EXTRACT FROM SCHIZANDRA |
| KR101619571B1 (en) * | 2013-12-03 | 2016-05-10 | 주식회사 바이오에프디엔씨 | Anti-aging Composition for Skin External Application Comprising Magnolia Placenta Culture Extracts |
| CN104546611A (en) * | 2015-01-14 | 2015-04-29 | 上海相宜本草化妆品股份有限公司 | Application of deoxyschizandrin in preparation of cosmetics or externally-applied medicines with functions of resisting light aging or removing acnes and diminishing inflammation |
| CN110037946A (en) * | 2019-06-04 | 2019-07-23 | 河南发发发生物科技有限公司 | A kind of black hair Hair-conditioning essential oil and preparation method thereof |
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