Matrine composition and application thereof

Abstract

The present application relates to the technical field of medicine. The present application provides a matrine composition and application thereof, the composition comprising the following molar parts of raw materials: matrine 1-10 parts, angelica sinensis lactone A 1-10 parts. When the composition of the present application is applied to HaCaT cells, compared with the use of matrine alone, it can more significantly inhibit the proliferation of keratinocytes and reduce the production of inflammatory factors. In a psoriasis-like mouse model, compared with the use of matrine alone, the composition can more effectively improve the psoriasis symptoms of mice and significantly enhance the therapeutic effect of matrine on psoriasis.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a matrine composition and application thereof. Background Art

[0002] Psoriasis, commonly known as psoriasis, is a chronic skin disease mediated by environmental factors, polygenic inheritance, and immune mechanisms. Its clinical manifestations include scaly erythema or plaques, with some patients also experiencing joint symptoms and nail abnormalities. Psoriasis not only impacts patients' quality of life but is also associated with the development of multiple conditions, including metabolic syndrome and cardiovascular disease. In severe cases, psoriasis can be life-threatening.

[0003] Oxymatrine (OMT) is a natural alkaloid extracted from Chinese herbs such as Sophora flavescens and Sophora flavescens. 15 H 24 N2O2. Studies have shown that matrine has a wide range of pharmacological activities, including anti-tumor, anti-fibrotic, antiviral, and anti-inflammatory effects. Multiple reports indicate that matrine has potential in the treatment of psoriasis. For example, Li N et al. found that matrine inhibited keratinocyte proliferation in BALB / c mice with imiquimod-induced psoriatic dermatitis, reduced PCNA+ and CD3+ cell infiltration in lesions, and decreased the expression of MyD88 protein and the inflammatory cytokines IL-12, IL-23, and IL-1β, thereby alleviating psoriatic dermatitis. Jiang WW et al. also demonstrated that matrine inhibited HaCaT cell proliferation in a dose- and time-dependent manner. When used in combination with acitretin, matrine induced cell cycle arrest and autophagy by activating the PI3K / Akt / mTOR pathway, resulting in a synergistic effect. However, current research has largely focused on the effects of individual components, lacking the exploration of combining matrine with other active ingredients to enhance its therapeutic efficacy. Summary of the Invention

[0004] The purpose of the present invention is to provide a matrine composition and its application, which significantly enhances the effect of matrine in treating psoriasis by adding compound ingredients.

[0005] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0006] The invention provides a matrine composition, which comprises the following raw materials in molar parts: 1-10 parts of matrine and 1-10 parts of angelica lactone A.

[0007] Preferably, the composition comprises the following raw materials in molar proportions: 2 to 8 parts of matrine and 2 to 8 parts of angelica lactone A.

[0008] Preferably, the composition comprises the following molar parts of raw materials: matrine 3-7 parts, ligustilide A 3-7 parts.

[0009] Preferably, the composition comprises the following molar parts of raw materials: matrine 4-6 parts, ligustilide A 4-5 parts.

[0010] The application further provides use of the composition in preparation of a medicine for preventing or treating psoriasis.

[0011] The application further provides a medicine for preventing or treating psoriasis, wherein the composition comprises the composition.

[0012] Preferably, the medicine is in the form of an injection.

[0013] The application provides a matrine composition and use thereof, wherein the composition comprises the following molar parts of raw materials: matrine 1-10 parts, ligustilide A 1-10 parts. When applied to HaCaT cells, the composition of the application can more significantly inhibit the proliferation of keratinocytes and reduce the production of inflammatory factors, as compared with matrine alone. In a psoriasis-like mouse model, the composition can more effectively improve the psoriasis symptoms of mice and significantly enhance the therapeutic effect of matrine on psoriasis, as compared with matrine alone. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 Figure 1 shows the effect of matrine on imiquimod-induced HaCaT cells, wherein A shows the effect of different concentrations of imiquimod on promoting the proliferation of HaCaT cells; and B shows the effect of matrine on imiquimod-induced HaCaT cells.

[0015] Figure 2 Figure 2 shows the effect of ligustilide A on the inhibition of HaCaT cell proliferation by matrine, wherein A shows the structure of ligustilide A; B shows the change in the viability of HaCaT cells after the addition of ligustilide A, as detected by CCK-8; and C shows the change in the number of HaCaT cells after the addition of ligustilide A, as shown by Giemsa staining.

[0016] Figure 3 Figure 3 shows the effect of ligustilide A on the inhibition of the secretion of inflammatory factors by HaCaT cells by matrine, wherein A shows the change in the inflammatory factor IL-17; and B shows the change in the inflammatory factor IL-23.

[0017] Figure 4 Figure 4 shows the effect of ligustilide A on the improvement of the symptoms of psoriasis mice by matrine, wherein A shows the change in the appearance of the back of a psoriasis mouse; and B shows HE staining of the skin. DETAILED DESCRIPTION

[0018] The invention provides a matrine composition, which comprises the following raw materials in molar parts: 1-10 parts of matrine and 1-10 parts of angelica lactone A.

[0019] In the present invention, the composition preferably comprises the following raw materials in molar proportions: 2 to 8 parts of matrine and 2 to 8 parts of angelica lactone A.

[0020] In the present invention, the composition preferably comprises the following raw materials in molar proportions: 3 to 7 parts of matrine and 3 to 7 parts of angelica lactone A.

[0021] In the present invention, the composition preferably comprises the following raw materials in molar proportions: 4-6 parts of matrine and 4-5 parts of angelica lactone A.

[0022] The present invention also provides use of the composition in preparing a medicament for preventing or treating psoriasis.

[0023] The present invention also provides a medicine for preventing or treating psoriasis, wherein the composition contains the composition.

[0024] In the present invention, the dosage form of the drug is an injection.

[0025] The technical solutions provided by the present invention are described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0026] Example

[0027] 1. Cell culture:

[0028] HaCaT cells (China Center for Type Culture Collection, Wuhan) were cultured at 37°C and 5% CO2 in DMEM medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin.

[0029] 2. CCK-8 cell activity assay:

[0030] Cell counting: When HaCaT cells reach a monolayer covering the bottom of the culture dish, trypsinize the cells for 10 minutes. Then, add 2 μL of trypan blue dye to 18 μL of cell suspension for staining. Count the cells using a cell counter and adjust the cell concentration to 6 × 10 4 / mL.

[0031] Cell plating: After counting, the cell suspension was seeded into a 96-well plate at a concentration of 6000 cells / well. PBS was added to the surrounding wells to reduce the edge effect. After standing for 5 minutes, the plate was transferred to a CO2 incubator for further culture.

[0032] Imiquimod modeling: After 12 hours of cell attachment, prepare imiquimod solutions at concentrations of 3.6 μg / mL, 7.8 μg / mL, 15.6 μg / mL, 31.2 μg / mL, 62.5 μg / mL, and 125 μg / mL. Aspirate the top layer of culture medium from each well and add 100 μL of culture medium containing imiquimod. For the blank control group, only complete culture medium was added. Six replicate wells were set up for each group and the cells were transferred to a CO2 incubator for further incubation for 24 hours.

[0033] Oxymatrine treatment: After cells adhered, oxymatrine treatment solutions were added at concentrations ranging from 7.5 mM to 45 mM. The model group was treated with culture medium containing imiquimod and cultured for another 24 hours.

[0034] Treatment with matrine and angelica lactone A: After adherence, different concentrations of angelica lactone A (2, 4, and 6) and 7.5 mM matrine were added. Six replicate wells were set up for each group, and the cells were transferred to a CO2 incubator and cultured for 24 hours.

[0035] Cell viability assay: After administration, the culture medium was removed, 100 μL of 10% Cell Counting Kit-8 solution was added, and the cells were incubated for 40 minutes. The absorbance was measured at 450 nm using a microplate reader.

[0036] 3. Giemsa staining:

[0037] HaCaT cells were seeded in 6-well plates (2×10 5 After cells adhered, imiquimod, 7.5 mM matrine, and 2, 4, or 6 picocyanine lactone A treatment solutions were added. After 24 hours, the medium was aspirated, the cells were washed with PBS, and after drying, 1 mL of methanol was added for 5 minutes to fix the cells. The methanol was then aspirated, and 1 mL of Giemsa stain was added for 30 minutes. The stain was diluted with PBS (1x concentration), and the cells were washed four times and photographed.

[0038] 4. Determination of inflammatory factors IL-17 and IL-23

[0039] Remove the kit from the refrigerator 30 minutes in advance and equilibrate to room temperature. Based on the number of samples and standards, prepare the required solutions and determine the number of strips. Add 100 μL of sample or standard solution to each well (add only standard or sample diluent to the blank control wells). Seal the wells with sealing tape and incubate at 37°C for 90 minutes (except for the blank control wells). After incubation, shake off all liquid in the wells. Add 350 μL of wash solution to each well, let it sit for 30 seconds, shake off all liquid, and pat dry on absorbent paper. Repeat four times. Then, add biotinylated antibody working solution (100 μL / well), incubate at 37°C for 60 minutes, and wash again four times. Next, add enzyme conjugate working solution (100 μL / well), incubate at 37°C for 30 minutes, and wash four times. Finally, add the color developing agent (100 μL / well), incubate in the dark for 20 minutes, add the stop solution (100 μL / well), mix well and quickly measure the OD value at 450 nm (within 5 minutes).

[0040] 5. Psoriasis mouse modeling and drug administration

[0041] All mice were housed in the GLP (Good Laboratory Practice) animal room of Lanzhou University. Fifty mice were randomly divided into a blank control group, a model group, a methotrexate group, a matrine group, and a matrine + angelica lactone A group, with 10 mice in each group.

[0042] Hair was removed from a 2cm×3cm area on the back of the mice. The control group was treated with 62.5mg of petrolatum, and the model group was treated with 62.5mg of imiquimod cream. The treatment groups were given methotrexate (7mg / kg), oxymatrine (25mg / kg), and oxymatrine plus angelica lactone A (25mg / kg + 6 doses) in addition to the model group. Imiquimod was applied in the morning and the corresponding drug treatment was given in the afternoon for a total of 7 days. After the treatment, photos were taken and samples were taken for HE staining (performed by Wuhan Sewell Company).

[0043] result

[0044] 1. Inhibitory effect of oxymatrine on imiquimod-induced HaCaT cell proliferation

[0045] The results of CCK-8 assay showed that compared with the blank control group, the viability of HaCaT cells treated with imiquimod (IMQ) was significantly increased, and different concentrations of imiquimod showed an increasing trend, among which the proliferation-promoting effect at concentrations of 3.6 μg / mL and 7.2 μg / mL was the most significant (see Figure 1 A), indicating that the HaCaT cell proliferation model was successfully established. Oxymatrine at a concentration of 7.5-45mM can significantly inhibit the proliferation-promoting effect of imiquimod on HaCaT cells. Within the concentration range studied, the increase in the concentration of oxymatrine and the inhibitory effect on HaCaT cell proliferation showed a clear dose-dependent relationship (see Figure 1 B) Curve fitting analysis revealed that the IC50 value of matrine for inhibiting HaCaT cell proliferation was 18.6 mM.

[0046] 2. Enhancement of the inhibitory effect of oxysporum flavescentis on HaCaT cell proliferation by oxysporum flavescentis lactone A

[0047] In this example, the concentration of oxymatrine was 7.5 mM as the basic concentration for subsequent studies. The experimental results showed that when different amounts of oxymatrine A (LeA) were added, namely 2 parts, 4 parts, 6 parts and 8 parts, the inhibitory effect of oxymatrine on HaCaT cells was gradually enhanced compared to the use of 7.5 mM oxymatrine alone, and the enhancement effect became more significant with the increase of oxymatrine A concentration (see Figure 2 B). When the amount of angelica lactone A is 4 parts, the IC50 value of oxymatrine can be achieved. Compared with the 18.6mM of oxymatrine used alone, the oxymatrine concentration can be reduced by 11mM. In addition, the Giemsa staining results further confirmed this point. Oxmatrine can inhibit the proliferation of HaCaT cells, and the inhibitory effect on HaCaT cells is more obvious after the addition of angelica lactone A (see Figure 2 C), With the addition of angelica lactone A, the number of HaCaT cells gradually decreased.

[0048] 3. The enhancing effect of angelica lactone A on the inhibitory effect of matrine on the inflammatory response of HaCaT cells

[0049] In this example, a concentration of 7.5 mM of matrine was selected for further study. The results showed that matrine could significantly reduce the levels of inflammatory factors IL-17 and IL-23 in HaCaT cells induced by imiquimod. After adding different amounts of angelica lactone A, the ability of matrine to reduce the level of inflammatory factors was further enhanced, and there was a significant difference (see Figure 3 ).

[0050] 4. Angelica lactone A enhances the effect of matrine on improving psoriasis-like mice

[0051] In animal experiments, compared with the blank group, mice in the imiquimod-treated group showed obvious scales on their backs and thickened keratin, which were typical symptoms of psoriasis. The positive control drug methotrexate (MTX) can significantly reduce the number of scales and thin the stratum corneum. Oxymatrine also showed a similar effect, significantly reducing the number of scales and the thickness of the stratum corneum. The combination of oxymatrine and angelica lactone A showed a more obvious effect than the oxymatrine alone group. The morphology of the back skin was close to that of the blank group, with almost no scales (see Figure 4 HE staining results showed that matrine could effectively reduce the thickness of the mouse back skin, and when used in combination with angelica lactone A, the effect of reducing skin thickness was more significant (see Figure 4B) In conclusion, angelica lactone A can significantly enhance the improvement effect of matrine on psoriasis-like mice.

[0052] As can be seen from the above examples, the present invention provides a matrine composition and its application, wherein the composition comprises the following raw materials in molar proportions: 1 to 10 parts of matrine and 1 to 10 parts of angelica lactone A. When applied to HaCaT cells, the composition of the present invention can more significantly inhibit the proliferation of keratinocytes and reduce the production of inflammatory factors compared to the use of matrine alone. In a psoriasis-like mouse model, the composition can more effectively improve the psoriasis symptoms of mice compared to the use of matrine alone, significantly enhancing the therapeutic effect of matrine on psoriasis.

[0053] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as within the scope of protection of the present invention.

Claims

1. A matrine composition, characterized in that: The composition is composed of the following raw materials in molar parts: 1-10 parts of oxymatrine and 1-10 parts of angelica lactone A.

2. The composition according to claim 1, characterized in that The composition is composed of the following raw materials in molar parts: 2-8 parts of oxymatrine and 2-8 parts of angelica lactone A.

3. The composition according to claim 2, characterized in that The composition is composed of the following raw materials in molar parts: 3-7 parts of oxymatrine and 3-7 parts of angelica lactone A.

4. The composition according to claim 3, characterized in that The composition is composed of the following raw materials in molar parts: 4-6 parts of oxymatrine and 4-5 parts of angelica lactone A.

5. Use of the composition according to any one of claims 1 to 4 in the preparation of a medicament for preventing or treating psoriasis.

6. A drug for preventing or treating psoriasis, characterized in that: The medicine contains the composition according to any one of claims 1 to 4, and the dosage form of the medicine is an injection.

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