Application of methyl cedryl ether in preparation of product for resisting skin photoaging

Abstract

The invention belongs to the technical field of medicines, and particularly provides a novel application of methyl cedryl ether in preparation of a product for resisting skin photoaging. Cell experiment evaluation proves that the methyl cedryl ether has no obvious influence on the activity of dermal Hs68 fibroblasts at the concentration of 12.5-100 mu M, and meanwhile, the methyl cedryl ether can obviously increase the content of hyaluronic acid in light-aged Hs68 cells induced by medium-wave ultraviolet rays (UVB), improve the mRNA expression level of COL1A2 and COL3A1, improve the mRNA expression level of HAS1 and HAS3 and reduce the mRNA expression level of MMP-1, MMP-3 and MMP-9. Therefore, the methyl cedryl ether provided by the invention can be applied to preparation of products for resisting skin photoaging.

Description

Technical Field

[0001] The invention belongs to the technical field of medicines and specifically discloses use of methyl cedarwood ether in preparing a product for resisting skin photoaging. Background Art

[0002] The skin is the largest organ in the human body and the first line of defense against external stimuli. It is often exposed to the external environment. Skin aging is affected by intrinsic factors (age, genetic composition, hormonal changes, etc.) and extrinsic factors (ultraviolet radiation, air pollution, etc.). About 80% of skin aging is caused by ultraviolet exposure. UVB has strong genotoxicity and has a sunburn ability about 1,000 times higher than UVA. Long-term exposure of the skin to UVB can cause erythema, sunburn, oxidative damage, apoptosis and inflammation, and even lead to skin cancer. In addition, skin aging can cause complex psychological problems (such as anxiety, depression, inferiority, etc.), which in turn affect people's quality of life. Skin photoaging has become a public health issue. At present, plant-derived flavor compounds have been shown to have beneficial effects in alleviating skin photoaging, becoming an important candidate substance for anti-photoaging damage.

[0003] Methyl cedryl ether, also known as cedryl methyl ether, can be synthesized from cedryl alcohol contained in cedar oil. As an aromatic ingredient, methyl cedryl ether is widely used in the cosmetics industry and in the production of spices. In recent years, research on the biological activity of methyl cedryl ether has mainly focused on its insecticidal and anti-obesity abilities. Existing studies have shown that methyl cedryl ether, as an inverse agonist, may have an inhibitory effect on ligand-independent α-adrenergic-like octopamine receptors in insects, thereby exerting insecticidal activity. In vivo studies have shown that methyl cedryl ether has the effect of preventing obesity and related metabolic diseases induced by a high-fat diet. However, the preventive and therapeutic effects of methyl cedryl ether on skin diseases such as photoaging have not been reported. Summary of the invention

[0004] The purpose of the present invention is to provide a new use of methyl cedarwood ether, specifically, to provide the use of methyl cedarwood ether in the preparation of anti-skin photoaging products.

[0005] In order to achieve the above object, the technical solution adopted by the present invention is:

[0006] Use of methyl cedarwood ether in preparing products for resisting skin photoaging.

[0007] Preferably, the methyl cedarwood ether has no significant effect on the viability of Hs68 cells at a concentration of 12.5-100 μM.

[0008] Preferably, the methyl cedarwood ether can increase the hyaluronic acid content in UVB-induced photoaged Hs68 cells.

[0009] Preferably, the methyl cedarwood ether can increase the mRNA expression levels of COL1A2 and COL3A1, increase the mRNA expression levels of HAS1 and HAS3, and reduce the mRNA expression levels of MMP-1, MMP-3 and MMP-9 in UVB-induced photoaging Hs68 cells.

[0010] The present invention also provides a health food composition for resisting skin photoaging, which comprises any one of the above-mentioned methyl cedarwood ethers and an auxiliary material acceptable to food.

[0011] Preferably, the health food composition has no significant effect on the viability of Hs68 cells at a concentration of 12.5-100 μM, can increase the hyaluronic acid content of photoaged cells, increase the mRNA expression levels of COL1A2 and COL3A1, increase the mRNA expression levels of HAS1 and HAS3, and reduce the mRNA expression levels of MMP-1, MMP-3 and MMP-9.

[0012] The present invention also provides a pharmaceutical composition for resisting skin photoaging, comprising any one of the above-mentioned methyl cedarwood ethers, and a pharmaceutically acceptable adjuvant or carrier.

[0013] Preferably, the medicine can be an oral solution, powder, tablet, capsule or injection.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention has discovered a new use of methyl cedryl ether. The results of cell experiments show that treating Hs68 cells irradiated with UVB at a concentration of 100 μM can significantly increase the hyaluronic acid content of photoaged dermal fibroblasts, increase the mRNA expression levels of COL1A2 and COL3A1, increase the mRNA expression levels of HAS1 and HAS3, and reduce the mRNA expression levels of MMP-1, MMP-3 and MMP-9, indicating that methyl cedryl ether can alleviate UVB-induced skin photoaging. The present invention provides a new use of methyl cedryl ether in the preparation of related health foods and medicines that have the effect of alleviating UVB-induced skin photoaging. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 This is the effect of methyl cedarwood ether on the viability of Hs68 cells.

[0017] Figure 2 This is the effect of methyl cedryl ether on the hyaluronic acid content in photoaged Hs68 cells. * p<0.05, ** p<0.01.

[0018] Figure 3This is the effect of methyl cedarwood ether on the mRNA expression levels of COL1A2 and COL3A1 in photoaged Hs68 cells. * p<0.05, ** p<0.01, *** p<0.001.

[0019] Figure 4 This is the effect of methyl cedarwood ether on the mRNA expression levels of HAS1 and HAS3 in photoaged Hs68 cells. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

[0020] Figure 5 This is the effect of methyl cedarwood ether on the mRNA expression levels of MMP-1, MMP-3 and MMP-9 in photoaged Hs68 cells. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.

[0021] Note: The present invention Figures 2 to 5 middle:

[0022] Control represents the control group, in which Hs68 cells were not irradiated with UVB;

[0023] UVB represents the model group, and Hs68 cells were irradiated with UVB;

[0024] MCE represents the methyl cedryl ether-treated group, in which Hs68 cells were treated with UVB irradiation and then with methyl cedryl ether;

[0025] CED represents the cedrol-treated group. Hs68 cells were treated with UVB and then given cedrol. DETAILED DESCRIPTION

[0026] The present invention is further described below in conjunction with the embodiments of the present invention and the accompanying drawings, but the protection scope of the present invention is not limited thereto.

[0027] Example 1

[0028] Effects of methyl cedarwood ether on the viability of dermal Hs68 fibroblasts

[0029] Hs68 cells were cultured in DMEM medium supplemented with 10% FBS and 1% penicillin-streptomycin solution at 37°C in the presence of 5% CO2. When cells were passaged, the cells were washed with PBS and then treated with 0.25% trypsin-ethylenediaminetetraacetic acid solution to detach the adherent cells. The cell viability was determined by the CCK8 method. After Hs68 cells were inoculated in a 96-well plate and cultured for 24 hours, they were treated with methyl cedar ether and cultured for another 24 hours. In addition, a control group containing only culture medium but no cells was set up. After incubation in the CCK8 reagent for 3 hours, the optical density (OD) of each well in the 96-well plate was measured at 450nm using a microplate reader. The formula for calculating cell viability (X, %) is as follows:

[0030]

[0031] Wherein: A2 is the OD value of each test well, A1 is the average OD value of the group without methyl cedar ether treatment, and A0 is the average OD value of the control group.

[0032] according to Figure 1 As shown, methyl cedarwood ether had no significant effect on the viability of Hs68 cells at concentrations of 12.5-100 μM.

[0033] Example 2

[0034] Effects of methyl cedryl ether on the content of hyaluronic acid in photoaged Hs68 cells

[0035] Hs68 cells were inoculated in 96-well plates and cultured for 24 hours to allow the cells to reach 50% to 60% confluence. The Hs68 cells to be treated were divided into 4 groups: control group, model group, methyl cedarwood ether treatment group and cedar alcohol treatment group, with 3 replicates in each group. Except for the control group, the other groups were given a single 5 mJ / cm 2 UVB radiation induces photoaging. The UVB lamp provides UVB radiation, and the radiation intensity is 0.126mW / cm 2 During UVB irradiation, cells were kept in PBS. The irradiation time (F, s) was calculated as follows:

[0036] F = a / b

[0037] Where: a is the radiation dose, mJ / cm 2 ; b is the irradiation intensity, mW / cm 2 .

[0038] After UVB irradiation, Hs68 cells in the methyl cedryl ether treatment group were treated with 100 μM methyl cedryl ether, and Hs68 cells in the cedryl alcohol treatment group were treated with 100 μM cedryl alcohol and cultured for 24 h. The hyaluronic acid content in the cells was determined using a hyaluronic acid ELISA kit, and the hyaluronic acid content was finally normalized using the total protein content of the cells.

[0039] according to Figure 2 As shown, compared with the control group, the hyaluronic acid content in the model group cells was significantly reduced, indicating that UVB irradiation promoted the degradation of cellular hyaluronic acid; methyl cedarwood ether could significantly increase the hyaluronic acid content in photoaged Hs68 cells, and the effect of methyl cedarwood ether on increasing the hyaluronic acid content in UVB-induced photoaged Hs68 cells was significantly better than cedrol.

[0040] Example 3

[0041] Effects of methyl cedarwood ether on the mRNA expression levels of COL1A2 and COL3A1 in photoaged Hs68 cells

[0042] The total RNA of Hs68 cells treated in groups in Example 2 was extracted with TRIzol and the RNA concentration was determined. TransScript one-step gDNA removal and cDNA synthesis SuperMix was used for cDNA synthesis. Reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) was performed using SuperReal fluorescent quantitative premix reagent enhanced version. The primer sequences used for RT-qPCR are shown in Table 1. The mRNA expression level of the target gene was normalized with the expression level of β-actin.

[0043] Table 1 Primer sequences used for RT-qPCR

[0044]

[0045] according to Figure 3 As shown, compared with the control group, the mRNA expression levels of COL1A2 and COL3A1 in the model group cells were significantly decreased; methyl cedarwood ether could significantly increase the mRNA expression levels of COL1A2 and COL3A1 in UVB-induced photoaging Hs68 cells, and the effect was significantly better than that of cedarol.

[0046] Example 4

[0047] Effects of methyl cedarwood ether on the mRNA expression levels of HAS1 and HAS3 in photoaged Hs68 cells

[0048] The total RNA of Hs68 cells treated in groups in Example 2 was extracted with TRIzol and the RNA concentration was determined. TransScript one-step gDNA removal and cDNA synthesis SuperMix was used for cDNA synthesis. Reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) was performed using SuperReal fluorescent quantitative premix reagent enhanced version. The primer sequences used for RT-qPCR are shown in Table 1. The mRNA expression level of the target gene was normalized with the expression level of β-actin.

[0049] according to Figure 4 As shown, compared with the control group, the mRNA expression levels of HAS1 and HAS3 in the model group cells were significantly decreased; methyl cedarwood ether could significantly increase the mRNA expression levels of HAS1 and HAS3 in UVB-induced photoaged Hs68 cells, and the effect was significantly better than that of cedarol.

[0050] Example 5

[0051] Effects of methyl cedarwood ether on the mRNA expression levels of MMP-1, MMP-3 and MMP-9 in photoaged Hs68 cells

[0052] The total RNA of Hs68 cells treated in groups in Example 2 was extracted with TRIzol and the RNA concentration was determined. TransScript one-step gDNA removal and cDNA synthesis SuperMix was used for cDNA synthesis. Reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) was performed using SuperReal fluorescent quantitative premix reagent enhanced version. The primer sequences used for RT-qPCR are shown in Table 1. The mRNA expression level of the target gene was normalized with the expression level of β-actin.

[0053] according to Figure 5 As shown in the results, compared with the control group, the mRNA expression levels of MMP-1, MMP-3 and MMP-9 in the model group cells were significantly increased; methyl cedarwood ether could significantly reduce the mRNA expression levels of MMP-1, MMP-3 and MMP-9 in UVB-induced photoaged Hs68 cells, and the effect was significantly better than that of cedarol.

[0054] The present invention is not limited to the above-mentioned embodiments. Without departing from the essential content of the present invention, any deformation, improvement and substitution that can be conceived by those skilled in the art shall fall within the scope of the present invention.

Claims

1. Use of methyl cedarwood ether in the preparation of anti-skin photoaging products.

2. The use according to claim 1, characterized in that The methyl cedarwood ether can alleviate UVB-induced skin photoaging.

3. The use according to claim 2, characterized in that: The methyl cedarwood ether had no significant effect on the viability of dermal Hs68 cells at a concentration of 12.5-100 μM.

4. The use according to claim 2, characterized in that: The methyl cedryl ether can increase the hyaluronic acid content in UVB-induced photoaging Hs68 cells.

5. The use according to claim 2, characterized in that: The methyl cedarwood ether can increase the mRNA expression levels of COL1A2 and COL3A1 in UVB-induced photoaging Hs68 cells, increase the mRNA expression levels of HAS1 and HAS3, and reduce the mRNA expression levels of MMP-1, MMP-3 and MMP-9.

6. The use according to claim 1, characterized in that: The product is a health food composition or a pharmaceutical composition.

7. A health food composition for resisting skin photoaging, characterized in that: The invention comprises the methyl cedarwood ether according to any one of claims 1 to 5, and an auxiliary material acceptable to food.

8. The health food composition according to claim 7, characterized in that The health food composition can have no significant effect on the viability of Hs68 cells at a concentration of 12.5-100 μM, increase the hyaluronic acid content of photoaged Hs68 cells, increase the mRNA expression levels of COL1A2 and COL3A1, increase the mRNA expression levels of HAS1 and HAS3, and reduce the mRNA expression levels of MMP-1, MMP-3 and MMP-9.

9. A pharmaceutical composition for resisting skin photoaging, characterized in that: The invention comprises the methyl cedarwood ether according to any one of claims 1 to 5, and a pharmaceutically acceptable auxiliary material or carrier.

10. The drug for resisting skin photoaging according to claim 9, characterized in that: The medicine can be oral solution, powder, tablet, capsule or injection.

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