Inflammatory cytokine inhibition device and inhibition method

Abstract

To inhibit inflammatory cytokine in skin or a cell in an inflammation state.SOLUTION: Light having a wavelength of 500 to 600 nm is emitted to skin by using an inflammatory cytokine inhibition device including an irradiation mechanism that emits the light having the wavelength of 500 to 600 nm.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to an apparatus and method for inhibiting inflammatory cytokines. [Background technology]

[0002] Cytokines are a general term for soluble proteins that carry a variety of intercellular signals within the body, and inflammatory cytokines are causative factors (inflammatory factors) that cause various inflammatory symptoms within the body.

[0003] There is a need for a method to suppress inflammatory cytokines in order to improve conventional skin inflammation. Patent Document 1 proposes an apparatus for suppressing the production of inflammatory cytokines using light irradiation. In the examples of Patent Document 1, light from a xenon discharge tube is passed through a bandpass filter with a central wavelength of 650 nm (half width 40 nm), and the transmitted light is irradiated onto a mouse ear. 5 days, 12 days, and 20 days after the start of light irradiation, a drug is applied to cause inflammation (mouse ear inflammation). When light irradiation was performed, the increase in ear thickness was reduced (swelling was suppressed) compared to when light irradiation was not performed. [Prior art documents] [Patent documents]

[0004] [Patent Document 1] JP 2012-147866 A Summary of the Invention [Problem to be solved by the invention]

[0005] However, the experiment shown in Patent Document 1 only confirmed preventive inflammation suppression, rather than an approach to skin or cells that were already in an inflamed state, and the suppressive effect was insufficient. An object of the present invention is to provide an apparatus and method for suppressing inflammatory cytokines, which can suppress inflammatory cytokines in skin and cells in an inflamed state. [Means for solving the problem]

[0006] The present inventors have found that irradiation with light having a wavelength of 500 to 600 nm is effective in suppressing inflammatory cytokines. That is, the present invention has the following aspects. [1] An inflammatory cytokine suppression device having an irradiation mechanism that irradiates light with a wavelength of 500 to 600 nm. [2] A method for suppressing inflammatory cytokines by irradiating the skin with light with a wavelength of 500 to 600 nm. [3] The light has an energy accumulation of 2.4 to 7.2 J / cm 2 The method for suppressing inflammatory cytokines described in [2], wherein irradiation is performed so as to Effect of the Invention

[0007] According to the present invention, it is possible to suppress inflammatory cytokines in skin or cells in an inflamed state. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008] The proinflammatory cytokine-suppressing device of the present invention has an irradiation mechanism that irradiates light with a wavelength of 500 to 600 nm. The irradiation mechanism is not particularly limited as long as it can irradiate the target with light having a wavelength in the range of 500 to 600 nm. The light irradiated to the target may contain light with a wavelength outside the range of 500 to 600 nm, provided that the effects of the present invention are not impaired. The light irradiated to the target may be cut off from light with a wavelength outside the range of 500 to 600 nm.

[0009] The method for suppressing inflammatory cytokines of the present invention is a method for irradiating the skin with light having a wavelength of 500 to 600 nm. It is preferable to irradiate the skin with light having a wavelength of 500 to 600 nm using the proinflammatory cytokine-suppressing device of the present invention.

[0010] As one embodiment of the irradiation mechanism of the inflammatory cytokine suppression device, an LED light having a dominant wavelength (main wavelength) of 500 to 600 nm can be used. The dominant wavelength is preferably 525 to 600 nm, and more preferably 550 to 580 nm. A commercially available green LED light may be used as the LED light. Generally, the dominant wavelength of green LED light is about 525 to 575 nm. By irradiating a target area of ​​the skin with light emitted from the LED light, inflammatory cytokines can be suppressed.

[0011] As another embodiment of the irradiation mechanism of the proinflammatory cytokine suppressing device, a combination of a phosphor that emits fluorescence with a wavelength of 500 to 600 nm and an excitation light source can be used. Examples of phosphors that emit fluorescence with a wavelength of 500 to 600 nm when irradiated with ultraviolet light from an excitation light source include zinc oxide phosphors, calcium phosphate phosphors, and magnesium titanate phosphors. Commercially available powder preparations containing phosphors may be used, such as Lumate G (a product of Sakai Chemical Industry Co., Ltd., a zinc oxide phosphor preparation), Lumate B (a product of Sakai Chemical Industry Co., Ltd., a calcium phosphate phosphor preparation), and Lumate R (a product of Sakai Chemical Industry Co., Ltd., a magnesium titanate phosphor preparation). By irradiating a target site on the skin with ultraviolet light while the fluorescent substance is present at the target site, inflammatory cytokines can be suppressed. Methods for making a fluorescent substance present at a target site on the skin include, for example, a method of scattering a powder formulation onto the target site on the skin, and a method of applying a dispersion in which the powder formulation is dispersed in a medium (e.g., a transparent gel) onto the target site on the skin.

[0012] Another embodiment of the irradiation mechanism of the inflammatory cytokine suppression device can be a combination of a light source that emits light in a wavelength range including 500 to 600 nm and a filter that transmits light within the wavelength range of 500 to 600 nm and does not transmit light outside the wavelength range of 500 to 600 nm. By irradiating a target site on the skin with light emitted from the light source through the filter, inflammatory cytokines can be suppressed.

[0013] In any embodiment, when the skin is irradiated with light having a wavelength of 500 to 600 nm, the accumulated energy amount is 2.4 to 7.2 J / cm 2 It is preferable to irradiate so that Accumulated energy (unit: J / cm 2 ) is the irradiation intensity (unit: W / m 2 It is calculated by multiplying the irradiation intensity by the irradiation time (unit: seconds). When the irradiation intensity is constant, the accumulated energy can be adjusted by changing the irradiation time.

[0014] When the irradiation mechanism is a combination of a phosphor and an excitation light source, the irradiation intensity can be adjusted by the amount of the phosphor. For example, a powder formulation containing a 15% by mass concentration of phosphor is applied to the target site on the skin at a depth of 10 cm 2 When present in an amount of 1 g per cm, the amount of phosphor present is 0.15 g / cm 2 It becomes. In this state, the excitation light source emits 1 mW / cm 2 When exposed to ultraviolet light of 0.15 g / cm (the same level as sunlight on a clear day), 2 0.3mW / cm from the phosphor 2 When the UV irradiation time is 200 minutes (12,000 seconds), the cumulative energy irradiated to the skin is 3.6 J / cm 2 It becomes. In addition, without changing the UV irradiation intensity, the presence of phosphor was reduced to 0.05 g / cm 2 When the change is made to 0.1 mW / cm, the phosphor emits 0.1 mW / cm 2 When the UV irradiation time is 200 minutes (12,000 seconds), the cumulative energy irradiated to the skin is 1.2 J / cm 2 It becomes.

[0015] According to the present invention, the expression of inflammatory cytokines can be suppressed by irradiating the skin with light having a wavelength of 500 to 600 nm. Specifically, it exerts a high inhibitory effect on inflammatory factors (prostaglandin E2, MCP-1, IL-8, etc.) expressed by external stress. For example, as shown in the examples below, when cells that have been irradiated with ultraviolet light to cause inflammation are irradiated with light of a wavelength of 500 to 600 nm, the expression of prostaglandin E2 is suppressed compared to when the light irradiation treatment is not performed. Prostaglandin E2 is a chemical mediator secreted from cells in which inflammation has occurred, and is involved in the production of inflammatory cytokines that enhance inflammation. The production of inflammatory cytokines is suppressed by suppressing the expression of prostaglandin E2. In addition, when cells that have been irradiated with ultraviolet light to create an inflamed state are subjected to light irradiation treatment, the expression of MCP-1 is suppressed compared to when light irradiation treatment is not performed. Chemokines such as MCP-1 are a type of inflammatory cytokine. Suppression of chemokine expression can improve chronic inflammation. In addition, when cells that have been irradiated with ultraviolet light to create an inflamed state are subjected to light irradiation treatment, the expression of IL-8 is suppressed compared to when light irradiation treatment is not performed. IL-8 is a type of inflammatory cytokine. Suppression of IL-8 expression results in an improvement in chronic inflammation. EXAMPLES

[0016] The present invention will be described in detail below with reference to examples, but the present invention is not limited to the following description.

[0017] <Example 1> [culture] Human adult epidermal keratinocytes were maintained in a petri dish at 37°C in Dulbecco's Modified Eagle Medium (DMEM, Thermo Fisher Scientific) supplemented with 10% by volume Fetal Bovine Serum (FBS, Thermo Fisher Scientific) and antibiotics such as 100 μ / mL penicillin and 100 μ / mL streptomycin. Then, 4 × 10 cells were cultured in a 12-well culture plate containing DMEM with 10% FBS by volume. 5The cells were seeded at 100x the concentration and cultured at 37°C for 24 hours.

[0018] [Light irradiation treatment] After the culture, the medium was replaced with phenol red-free DMEM supplemented with 1% by volume of FBS. An LED UV light (THORLABS product name "Mounted LED M310L1") emitting ultraviolet B rays (UVB) was placed directly under the culture plate, and UVB was irradiated at 200 mJ / cm. 2 The mice were irradiated under the above conditions to serve as an inflammation model. Thereafter, the cells of the inflammation model were irradiated with green light under the conditions shown in Table 1 using a green LED light (THORLABS product name "Mounted LED M565L3", dominant wavelength (main wavelength) 565 nm).

[0019] [Inflammatory factor quantification] After the light irradiation treatment was completed (for the control, the same time as the green light irradiation time had elapsed after UVB irradiation without green light irradiation), the mice were left at 37°C, and after 24 hours, the supernatant was sampled, and each inflammatory factor in the obtained sample was evaluated by ELISA according to a predetermined protocol. The results are shown in Table 1 (the same applies below). Prostaglandin E2 was measured using PGE2 Multi-Format ELISA Kits (K051-H1) from ARBOR ASSAYS. MCP-1 was measured using R&D's Human CCL2 / MCP-1 Quantikine ELISA Kit (DCP00). IL-8 was measured using the Human IL-8 (CXCL8) ELISA Kit (AKH-IL8) from Fujifilm Wako Shibayagi. The expression level of each inflammatory factor in each sample irradiated with green light was calculated as a relative expression rate (unit: %), based on the expression level (100%) of each inflammatory factor in the control irradiated with UVB only. The lower the relative expression rate, the greater the effect of suppressing the expression of inflammatory factors. [Evaluation of expression suppression effect] The effect of suppressing the expression of inflammatory factors was evaluated according to the following criteria. ◎: Relative expression rate is 74% or less. ○: Relative expression rate is more than 74% and less than 86%. △: Relative expression rate >86% and ≤99%. ×: Relative expression rate >99%.

[0020] <Examples 2 to 5> In Example 1, light irradiation treatment was carried out in the same manner as in Example 1, except that the irradiation conditions were changed as shown in Table 1, and the expression-suppressing effect was evaluated.

[0021] <Example 6> In this example, a blue LED light (THORLABS product name "Mounted LED M470L5", dominant wavelength 470 nm) was used instead of the green LED light. Energy accumulation: 3.6J / cm 2 As a result, light irradiation treatment was carried out in the same manner as in Example 1, and the expression suppression effect was evaluated.

[0022] <Example 7> In this example, a red LED light (THORLABS product name "Mounted LED M625L4", dominant wavelength 625 nm) was used instead of the green LED light. Energy accumulation: 3.6J / cm 2 As a result, light irradiation treatment was carried out in the same manner as in Example 1, and the expression suppression effect was evaluated.

[0023] [Table 1]

[0024] As shown by the results in Table 1, the expression of inflammatory factors was suppressed by irradiating epidermal keratinocytes in an inflamed state with green light. The results in Table 1 show that irradiating inflamed skin with green light can suppress the expression of inflammatory cytokines.

Claims

1. An inflammatory cytokine suppression device having an irradiation mechanism for irradiating light with a wavelength of 500 to 600 nm.

2. A method for suppressing inflammatory cytokines, comprising irradiating the skin with light having a wavelength of 500 to 600 nm.

3. The light has an energy accumulation amount of 2.4 to 7.2 J / cm ２ The method for suppressing inflammatory cytokines according to claim 2 , wherein the irradiation is performed so as to give a total concentration of 1000 ppm or more.

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