Application of stable silicic acid aqueous solution in preparation of chloasma treatment product

Abstract

The invention relates to an application of a stable silicic acid aqueous solution in preparation of a product for treating chloasma, and the mass concentration of the stable silicic acid aqueous solution is 0.5-2%. As an active component, the stable silicic acid aqueous solution can treat chloasma at the mass concentration of 0.5%-2%. Meanwhile, the stable silicic acid aqueous solution is low in raw material cost, environment-friendly and safe, and can be used as an alternative choice of a medicine for treating chloasma, which has a good curative effect and is safe.

Description

TECHNICAL FIELD

[0001] The application relates to the technical field of biological medicines, in particular to application of a stable silicic acid aqueous solution in preparation of a product for treating chloasma. BACKGROUND

[0002] Chloasma is a common acquired pigmentation skin disease, and is generally located in the exposed parts of the face. The disease mainly affects women, and 10% of chloasma patients are men. The general clinical manifestation of chloasma is symmetrical net-like pigmentation, and the color is different in depth, and the edge is clear. Progesterone can induce the formation of chloasma through various ways. On the one hand, it can stimulate the activity of melanocytes, and increase the synthesis of melanin. Specifically, progesterone can affect the expression and activity of melanin synthesis related enzymes (such as tyrosinase), and promote the transformation of melanin precursor substances into melanin. On the other hand, progesterone can also affect the microenvironment of the skin, for example, change the endocrine and metabolic state of the skin, and promote the production of inflammatory factors, which can further stimulate melanocytes, and at the same time affect the transport and distribution of melanin bodies, and finally lead to increased pigmentation.

[0003] Current research shows that the pathogenesis of chloasma is relatively complex and diverse. Simply speaking, there are several aspects: (1) chloasma is caused by abnormal metabolism of melanin, which is the main pathogenesis of the disease. Therefore, one of the important means to reduce the formation of melanin is to inhibit the activity of tyrosinase in the body. (2) The number of superoxide free radicals in the body increases due to various reasons, and the activity of SOD decreases. When the number of oxygen free radicals in the body increases, even reaches excessive level, and the corresponding antioxidant enzyme system, such as SOD, shows a downward trend, that is, the content of SOD cannot be proportional to the content of peroxidized lipid (LPO), which will cause a large accumulation of peroxidized lipid in the body, so that the membrane lipid will be peroxidized, and thus the structure and corresponding physiological function of the membrane will be damaged, which will cause various diseases, such as aging and pigmentation.

[0004] Therefore, it is necessary to find a chloasma treatment topical external medicine with low raw material cost, and environmental protection and safety. SUMMARY

[0005] The purpose of the present application is to provide a substitute drug for the current chloasma treatment topical external medicine, and to prepare a chloasma treatment product by using a stable silicic acid aqueous solution.

[0006] In order to achieve the above purpose, the application provides application of a stable silicic acid aqueous solution in preparation of a chloasma treatment product, wherein the mass concentration of the stable silicic acid aqueous solution is 0.5% to 2%.

[0007] Further, when used externally, the stable silicic acid aqueous solution is dispersed in an ointment base as an active ingredient.

[0008] Further, the present application provides a product for treating chloasma, the active ingredient of which is a stable silicic acid aqueous solution with a mass concentration of 0.5% to 2%, and the product is a medical device or a drug.

[0009] Based on the above technical solution, the present application has the following advantages:

[0010] The stable silicic acid aqueous solution as an active ingredient can treat chloasma at a mass concentration of 0.5% to 2%. Meanwhile, the raw material of the stable silicic acid aqueous solution is low in cost and environmentally friendly and safe, and can be used as a safe and effective alternative for treating chloasma. BRIEF DESCRIPTION OF DRAWINGS

[0011] The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiments of the application and serve to explain the principles of the application. In the drawings:

[0012] Figure 1 HE staining diagram of the skin tissue on the back of a chloasma mouse;

[0013] Figure 2 SOD content diagram of the skin on the back of a chloasma mouse;

[0014] Figure 3 MDA content diagram of the skin on the back of a chloasma mouse. DETAILED DESCRIPTION

[0015] The technical solution of the present application will be further described below with reference to the accompanying drawings and examples.

[0016] The present application provides an application of a stable silicic acid aqueous solution in preparing a product for treating chloasma, wherein the mass concentration of the stable silicic acid aqueous solution is 0.5% to 2%.

[0017] It should be noted that the stable silicic acid aqueous solution in the present application is a stable stable silicic acid aqueous solution, wherein the stable silicic acid aqueous solution refers to a stable solution without polymerization, and the main components include monosilicic acid (H4SiO4), metasilicic acid (H2SiO3) and hydrogen silicate (HSiO3 - In the present application, the mass concentration of 0.5% to 2% refers to the total concentration of the main components being 0.5% to 2%. In the present application, in order to simply represent the stable silicic acid aqueous solution, metasilicic acid (H2SiO3) is used to represent the stable silicic acid aqueous solution in the following examples and the diagrams in the accompanying drawings, which is not a substitute for metasilicic acid as a single component.

[0018] Currently, researchers have conducted extensive experiments and explorations in animal models of melasma to ensure their accuracy and reliability. Suitable animals for melasma modeling include HRM-2 hairless mice, BALB / C mice, DBA / 2 mice, and brown-yellow guinea pigs. The establishment of these models not only helps to better understand the pathogenesis of melasma but also provides important references for developing treatments and drugs.

[0019] This experiment will use female BALB / c mice as experimental animals. Currently, there are three generally accepted modeling methods: ultraviolet (UV) irradiation, progesterone irradiation, and a combination of UV irradiation and progesterone irradiation. This experiment will use the progesterone irradiation method: progesterone (20 mg / kg) will be injected into the flexor muscles of the hind leg of mice once daily, with alternating injections in the hind legs, for one month. Drug administration will begin two weeks after modeling. Extensive data shows that oxygen free radicals are important substances leading to melanin deposition. The normal human body possesses a strong antioxidant defense system, with key molecules such as superoxide dismutase (SOD) effectively protecting cells from oxygen free radical damage. However, there is an imbalance between oxidation and antioxidant reactions in patients with melasma. Studies have found that the activities of SOD and catalase (CAT) in the blood of these patients are significantly reduced, while the levels of lipid peroxides (LPO) and malondialdehyde (MDA) are significantly increased. This imbalance exacerbates free radical damage to cells, thereby accelerating the disease progression. This finding underscores the importance of regulating oxidative stress and maintaining antioxidant balance in the treatment of melasma. Therefore, this study will measure SOD activity, MDA content, and GSH-PX content to evaluate the treatment efficacy.

[0020] (a) Experimental materials

[0021] Laboratory animals: 30 female SPF-grade BALB / c mice, 6 weeks old, weighing 16-20 grams, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. They were housed separately in an SPF-grade animal room at a constant temperature of 22±2℃, with 12 hours of continuous day and night.

[0022] (II) Experimental Methods

[0023] Experimental grouping: Thirty BALB / c mice were randomly divided into a control group (Control), a model group (Model), a low-dose stable silicic acid aqueous solution group (0.5%), a medium-dose stable silicic acid aqueous solution group (1%), a high-dose stable silicic acid aqueous solution group (2%), and a positive control group (Calcipotrio), with 5 mice in each group. The mice were acclimatized for two days.

[0024] Mouse modeling: Every day at 2 PM, mice in the model group, low-dose stable silicic acid aqueous solution group (0.5%), medium-dose stable silicic acid aqueous solution group (1%), high-dose stable silicic acid aqueous solution group (2%), and positive control group (Calcipotrio) were injected with progesterone (20 mg / kg) into the flexor muscle of the hind leg root once a day. The injection was carried out alternately in the hind legs of the mice for 30 consecutive days.

[0025] Treatment with the test drug: Mice were treated with the test drug 15 days after modeling, followed by intragastric administration during the modeling process. The low-dose (0.5%), medium-dose (1%), and high-dose (2%) groups of stable silicic acid aqueous solution were each administered 100 μL of the corresponding concentration of the test drug. The positive control group was administered 100 μL of tranexamic acid at a concentration of 5 mg / mL.

[0026] Pathological examination: HE staining was used to observe pathological changes in mice with chloasma-like skin. Skin from the same area on the back of the mice was fixed in 4% paraformaldehyde. After 24 hours, tissue dehydration, paraffin embedding, paraffin sectioning, and HE staining were performed to observe epidermal thickness, keratinocyte proliferation, and the degree of inflammatory cell infiltration.

[0027] HE staining procedure steps:

[0028] (1) Dehydration: Place the fixed skin tissue in a dehydration box and then place it in a gradient of ethanol from low to high (75% ethanol, 90% ethanol, 90% ethanol, 100% ethanol I, 100% ethanol II, 100% ethanol III for 1 hour each.

[0029] (2) Transparency: Xylene I, Xylene II, and Xylene III, each transparent for 45 minutes.

[0030] (3) Wax impregnation: Paraffin I, Paraffin II, Paraffin III, 1 hour each.

[0031] (4) Embedding: After the skin tissue is immersed in paraffin, it is placed vertically in the embedding frame and liquid paraffin is slowly poured into the frame. After the paraffin solidifies, the embedded paraffin block is removed and corrected and marked.

[0032] (5) Sectioning: First, remove excess wax from the surface of the wax block. When the tissue is about to be exposed, place it in the microtome and cut it into 5 μm sections. Then, gently place the specimen in 42℃ warm water to flatten it. Use a glass slide to lift it out and dry it at 60℃.

[0033] (6) Dewaxing and hydration: Place the slices in a 60℃ oven and dry for 1 hour. Immerse the slices in xylene I and xylene II for 15 min each, then in 100% alcohol, 95% alcohol, 90% alcohol, 80% alcohol and 70% alcohol for 5 min each, and then in distilled water for 3 min.

[0034] (7) Staining: Hematoxylin staining solution for 5 min.

[0035] (8) Washing: Rinse with running tap water for about 15 minutes to turn the slices blue (or place them in alkaline water), but be careful not to let the water flow too strong to prevent the slices from falling off.

[0036] (9) Differentiation: Place the slice in 1% hydrochloric acid ethanol solution to decolorize. After about 2 seconds, the slice turns red and the color becomes lighter.

[0037] (10) Rinse: Place the slices in running tap water to restore their blue color.

[0038] (11) Dehydration I: Slices are placed in 50% ethanol → 70% ethanol → 80% ethanol for 3-5 min each.

[0039] (12) Counterstaining: Stain with 0.5% eosin ethanol solution for 1 min.

[0040] (13) Dehydration II: Wash away excess red color by immersing the slices in 95% ethanol, then immerse them in anhydrous ethanol for 3-5 minutes. Finally, blot away excess ethanol with absorbent paper.

[0041] (14) Transparency: Place the slices in xylene I and II for 3-5 min each.

[0042] (15) Sealing: Seal with neutral resin.

[0043] (2) Operation table

[0044]

[0045] MDA content detection:

[0046] (1) Sample processing is the same as SOD

[0047] (2) Operation table

[0048]

[0049] Cover the centrifuge tubes, poke a small hole in the cover with a needle, vortex mix, and incubate in a 95°C water bath (or boil uncovered) for 40 minutes. After incubation, cool under running water, then centrifuge at 3500–4000 rpm for 10 minutes. Collect the supernatant and measure the absorbance of each tube at 532 nm.

[0050] (III) Experimental Results

[0051] This experiment used BALB / c mice to investigate the effects of different concentrations of the test drug on melasma, and the following results were obtained.

[0052] 1. Effects of different concentrations of the test drug applied topically on the histopathology of skin tissue in mice with melasma.

[0053] Compared with the normal control group, the model group showed significantly thickened epidermis, abnormally elongated basal "pegs," inflammatory cell infiltration, and deformed appendages due to compression. Compared with the model group, the epidermal thickening was reduced in the stable silica aqueous solution intervention group, the abnormal morphology of the basal "pegs" was repaired, and inflammatory cell infiltration was significantly alleviated. Among them, the epidermal thickness in the 1% stable silica aqueous solution group was close to normal, the morphology of the pegs was regular, and the dermal inflammation / structural compression was significantly relieved. The positive control group showed epidermal thickness and peg morphology close to the normal control group, with the most significant pathological improvement.

[0054] 2. Effects of different concentrations of the test drug on serum and skin SOD levels in mice with melasma

[0055] After treatment, compared with the blank control group (135.2 U / mgprot), the SOD content in the skin of mice in the model group was significantly reduced (84.1 U / mgprot). The SOD content in mice treated with stable silicic acid aqueous solution and tranexamic acid was reduced to varying degrees compared with the blank control group. Specifically, the SOD content in mice treated with 1% stable silicic acid aqueous solution (106.7 U / mgprot) was significantly higher than that in the model group, as was the SOD content in mice treated with 2% stable silicic acid aqueous solution (103.7 U / mgprot). In the positive control group, the SOD content in mice treated with tranexamic acid (97.3 U / mgprot) was significantly higher than that in the model group. These results indicate that 1% stable silicic acid aqueous solution, 2% stable silicic acid aqueous solution, and the positive control drug tranexamic acid can all increase the SOD content in the skin of mice with melasma.

[0056] 3. Effects of different concentrations of the test drug on serum and skin MDA levels in mice with melasma.

[0057] After treatment, compared with the blank control group (7.1 nmol / mg prot), the MDA content in the skin of mice in the model group (4.4 nmol / mg prot) was significantly reduced. MDA content in mice treated with stable silicic acid aqueous solution and tranexamic acid showed varying degrees of reduction compared with the blank control group. Specifically, the MDA content in the skin of mice treated with 0.5% stable silicic acid aqueous solution (6.6 nmol / mg prot) was significantly higher than that in the model group, while the MDA content in the skin of mice treated with 1% stable silicic acid aqueous solution (5.3 nmol / mg prot) and the positive control tranexamic acid (5.1 nmol / mg prot) was increased compared with the model group. These results indicate that 0.5% stable silicic acid aqueous solution, 1% stable silicic acid aqueous solution, and the positive control tranexamic acid can all increase the SOD content in the skin of mice with melasma.

[0058] This experiment investigated the effects of different concentrations of the test drug on a mouse model of melasma through animal experiments. Pathological results showed that three different concentrations of stable silicic acid aqueous solutions could alleviate the epidermal thickness and morphological structural variations induced by progesterone in mice with melasma, improving pathological structural characteristics, with the 1% stable silicic acid aqueous solution showing the most significant effect. The detection of SOD and MDA levels in mouse skin indicated that different concentrations of stable silicic acid aqueous solutions affected the SOD and MDA levels in mouse skin. The results of 1% stable silicic acid aqueous solution treatment were consistent with those of mice treated with the positive control drug tranexamic acid, simultaneously increasing the SOD and MDA levels in the skin of mice with melasma.

[0059] Therefore, the above experimental results show that stabilized silicic acid aqueous solution can alleviate symptoms in mouse melasma model, especially 1% concentration stabilized silicic acid aqueous solution can alleviate symptoms in mouse melasma model, and its effect is close to that of positive drug tranexamic acid treatment.

[0060] Furthermore, when used topically, the stabilized silica aqueous solution is dispersed as an active ingredient in the ointment matrix to obtain a more suitable application method.

[0061] Furthermore, the present invention provides a product for treating melasma, wherein the active ingredient of the product is a stable silicic acid aqueous solution with a mass concentration of 0.5% to 2%, and the product is a medical device or a drug.

[0062] The stabilized silicic acid aqueous solution of this invention, as the active ingredient, can treat melasma at a mass concentration of 0.5% to 2%. Furthermore, the raw materials for the stabilized silicic acid aqueous solution are inexpensive, environmentally friendly, and safe, making it a viable alternative to other medications for treating melasma with good efficacy.

[0063] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them; although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications can still be made to the specific implementation of the present invention or equivalent substitutions can be made to some technical features without departing from the spirit of the technical solutions of the present invention, and all such modifications and substitutions should be covered within the scope of the technical solutions claimed in the present invention.

Claims

1. The application of a stable silicic acid aqueous solution in the preparation of products for treating melasma, wherein, The mass concentration of the stabilized silicic acid aqueous solution is between 0.5% and 2%.

2. The application according to claim 1, characterized in that: When used topically, the stabilized silica aqueous solution is dispersed as an active ingredient in the ointment base.

3. A product characterized in that, The product is used to treat melasma. The active ingredient of the product is a stable silicic acid aqueous solution with a mass concentration of 0.5% to 2%. The product is a medical device or a drug.

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