Application and extraction method of prinsepia utilis royle polysaccharide

Abstract

The invention discloses application and an extraction method of prinsepia utilis royle polysaccharide, which comprises the following steps: extracting prinsepia utilis royle residues with 95% ethanol at 85 DEG C, filtering, extracting the residues with deionized water at 85-95 DEG C for 2-3 times, merging extracts, performing alcohol precipitation with 90%-95% ethanol at 2-6 DEG C, removing protein through a Sevage method, centrifugally collecting supernate, and freeze-drying to obtain the prinsepia utilis royle polysaccharide. And finally determining the content of the prinsepia utilis royle polysaccharide by using an anthrone-sulfuric acid colorimetric method, wherein the obtained prinsepia utilis royle polysaccharide is used for treating and improving skin lesion of psoriasis. Tests prove that prinsepia utilis royle polysaccharide can significantly improve psoriasis skin lesion, reduce inflammatory factor expression, enhance skin barrier function and increase expression of epidermal related protein, and is helpful for relieving symptoms of psoriasis patients and improving skin conditions.

Description

Technical Field

[0001] The invention belongs to the field of natural active ingredient extraction and application, and particularly relates to an application of prickle fruit polysaccharide and an extraction method thereof. Background Art

[0002] Prinia prickly ash, also known as pricking thorn, flat-core wood, and oil-making fruit, is an evergreen shrub belonging to the genus Flat-core Wood in the Rosaceae family. The kernels of Prinia prickly ash are traditionally used for oil extraction, and the residue is called Prinia prickly ash oil cake. Prinia prickly ash oil cake is rich in polysaccharides, which is one of its main active substances.

[0003] Principal prickle polysaccharide has multiple physiological functions, including anti-cancer, anti-obesity, blood sugar control, lowering cholesterol and blood lipids, anti-tumor and immune enhancement. In addition, Principal prickle polysaccharide also has the function of regulating cell growth and aging, controlling cell division and differentiation. However, no studies have reported the role of Principal prickle polysaccharide in improving psoriasis lesions.

[0004] Psoriasis is a common chronic, recurrent, inflammatory skin disease, mainly manifested by symptoms such as erythema, scaling and skin thickening. The exact cause of the disease is still unclear, and it is currently believed that its occurrence is closely related to immune system abnormalities. Existing treatments include topical medications, systemic treatments and phototherapy, but there are still problems such as unstable efficacy and large side effects. Therefore, finding new and effective treatments has become one of the research hotspots.

[0005] Extracting pharmacologically active ingredients from natural plants and using them to treat diseases is an important direction in current drug research and development. Although polysaccharides from the fruit of the thorn tree have been shown to have multiple physiological functions, their application in the treatment of skin diseases, especially psoriasis, has not been systematically studied. There is still a lot of work to be done in this field in order to find safer and more effective treatments.

[0006] In this context, the inventors of this patent focused on the extraction of polysaccharides from Prunus prickle fruit and its medical applications, especially its potential for treating skin diseases, and further revealed its potential health benefits through scientific experiments and research.

[0007] Therefore, the present invention provides an application of Principis utilis polysaccharide and an extraction method thereof. Summary of the invention

[0008] In order to solve the above technical problems, the present invention proposes an application of Principia principe polysaccharide and an extraction method thereof, indicating that Principia principe polysaccharide has significant clinical application prospects, especially in the treatment of psoriasis-related skin diseases.

[0009] In order to achieve the above technical effects, the present invention is implemented by the following technical solutions:

[0010] An application of Principessa principe polysaccharide, which is used for treating and improving psoriasis skin lesions; the Principessa principe polysaccharide has obvious effects in reducing scales, erythema and infiltration and thickening, effectively reduces transepidermal water loss of the skin, increases the moisture content of the skin, plays a positive role in enhancing the skin barrier function, significantly reduces the expression of inflammatory cytokines, has good anti-inflammatory effects, increases the expression of epidermal barrier-related proteins, and helps to improve the skin health of patients with psoriasis.

[0011] Another object of the present invention is to provide a method for extracting polysaccharides from Principal Insects, characterized in that it comprises the following steps:

[0012] The residue after oil is squeezed from the kernel of the prickle fruit is extracted 2-3 times with 95% ethanol at 80-95°C; the ethanol extract is cooled to room temperature and filtered, and the residue is extracted 2-3 times with deionized water at 85-95°C; the extract solution is concentrated and combined under reduced pressure, and then alcohol precipitation is performed with 90%-95% ethanol at 2-6°C; the protein in the precipitate is removed by Sevage method, and the supernatant is collected by centrifugation; the supernatant is freeze-dried to obtain the prickle fruit polysaccharide.

[0013] The beneficial effects of the present invention are:

[0014] 1. The experiment showed that the polysaccharide from Principia utilis had a significant effect in improving psoriasis skin lesions. Compared with the model group, the PASI score of mice in the polysaccharide group was significantly reduced, indicating its effectiveness in reducing scaling, erythema and infiltration thickening.

[0015] 2. The experimental results show that the polysaccharide of Principesto utilis can effectively reduce the transepidermal water loss of the skin and increase the moisture content of the skin, which indicates that it has a positive effect on enhancing the skin barrier function;

[0016] 3. The polysaccharide from Psoralea corylifolia showed good anti-inflammatory effects by reducing the expression of inflammatory cytokines (such as IL-17A, IL-23, IL-6, IL-1β, TNF-β). The reduction of these cytokines helps to alleviate the inflammatory response caused by psoriasis.

[0017] 4. The present invention increases the expression of epidermal barrier-related proteins, which are essential for maintaining the structure and function of the skin and help improve the skin health of psoriasis patients. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings required for describing the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other accompanying drawings can be obtained based on these accompanying drawings without paying creative work.

[0019] Figure 1 This is a diagram showing the changes in skin lesions of mice in different treatment groups in the present invention;

[0020] Figure 2 This is a trend chart of the PASI score of mice of the present invention;

[0021] Figure 3 This is a graph showing the determination of epidermal water loss (TEWL) values ​​in mice according to the present invention;

[0022] Figure 4 This is a graph showing the measurement of mouse epidermal moisture value according to the present invention;

[0023] Figure 5 This is a graph showing the content of IL-17A detected by ELISA of the present invention;

[0024] Figure 6 This is a graph showing the content of IL-23 detected by ELISA of the present invention;

[0025] Figure 7 This is a graph showing the content of IL-6 detected by ELISA of the present invention;

[0026] Figure 8 This is a graph showing the content of TGF-β detected by ELISA of the present invention;

[0027] Fig. 9 This is a graph showing the content of IL-1β detected by ELISA of the present invention;

[0028] Fig.10 This is a graph showing the content of ceramide detected by ELISA of the present invention;

[0029] Fig.11 This is a graph showing the cholesterol content detected by ELISA of the present invention;

[0030] Fig.12 This is a graph showing the expression level of epidermal barrier-related protein Filaggrin detected by WB of the present invention;

[0031] Fig.13 This is a graph showing the expression level of Loricrin, an epidermal barrier-related protein, detected by WB of the present invention;

[0032] Fig.14 This is a graph showing the expression level of the epidermal barrier-related protein Involucrin detected by WB of the present invention; DETAILED DESCRIPTION

[0033] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0034] Example 1

[0035] See also Figures 1 to 14 As shown, a method for extracting polysaccharides from Principal Utility Fruit is characterized by comprising the following steps:

[0036] S1. Extracting the residue after oil extraction from the pruning nut kernels with 95% ethanol for 3 times at 85° C. for 2 hours;

[0037] S2, cooling the ethanol extract to room temperature and filtering it, and extracting the residue with deionized water three times at 90°C for 2 hours;

[0038] S3, concentrating the combined extract solution under reduced pressure, and then performing alcohol precipitation with 95% ethanol at 4°C;

[0039] S4, remove the protein in the precipitate by Sevage method, and collect the supernatant by centrifugation;

[0040] S5. The supernatant was freeze-dried to obtain the polysaccharide of Psoralea utilis. The content of the polysaccharide was determined by anthrone-sulfuric acid colorimetry at a wavelength of 582 nm using D-glucose as a standard, and the result was 32.2%.

[0041] Example 2

[0042] Biological activity test of polysaccharide from Prinus utilis:

[0043] 1. Experimental animals: 6-8 weeks old SPF female C57 mice were selected. After 7 days of adaptive feeding, 62.5 mg of 5% imiquimod cream was applied to the depilated skin of the mice for 10 days to construct a psoriasis mouse model.

[0044] 2. Treatment group: During the modeling process, the polysaccharide of Principia utilis was applied to the mouse skin, and the positive control drug was calcipotriol cream.

[0045] 3. Experimental content:

[0046] (1) Skin lesion scoring: Photos of mouse skin lesions were taken daily. According to the PASI scoring standard, the degree of erythema, scaling, and infiltration thickening of the mouse skin lesions was scored 0-4. The total score was obtained by adding the three scores.

[0047] (2) Evaluation of epidermal barrier function: Determination of skin TEWL and moisture value.

[0048] (3) Morphological observation: The dorsal skin of the mice was taken after death and stained with HE to observe the morphological changes.

[0049] (4) Cytokine detection: ELISA was used to detect the contents of ceramide, cholesterol, IL-17A, IL-23, IL-6, TGF-β, and IL-1β.

[0050] 4. Experimental results:

[0051] (1) Changes in psoriasis lesions such as Figure 1 shown.

[0052] (2) PASI score of mice

[0053] The skin lesions of the experimental mice were photographed daily. According to the PASI scoring standard, scaling, infiltration and erythema were evaluated on a scale of 0-4: 0 for no obvious symptoms; 1 for mild symptoms, 2 for moderate symptoms, 3 for severe symptoms, and 4 for extremely severe symptoms. The scaling, thickness and erythema scores of each group of mice were recorded and statistically analyzed daily. The total score was calculated by adding the three scores, and a PASI score trend chart was drawn to dynamically observe the changes in the skin lesions of the mice. Figure 2 As shown in the results, the PASI score of the Primulella utilis group was significantly lower than that of the model group (P<0.05), but had no significant difference with the calcipotriol group (P>0.05).

[0054] (3) Determination of epidermal water loss in mice

[0055] After measuring TEWL with a non-invasive skin detection probe every day, 100 mg / mL of Principia utilis polysaccharide and 40-50 mg of positive drugs were applied for smear administration. TEWL values ​​were measured again 4 hours after administration. Figure 3 As shown in the results, the TEWL value of the Principia utilis group was significantly lower than that of the model group (P<0.05), but had no significant difference with the calcipotriol group (P>0.05).

[0056] (4) Determination of mouse epidermal moisture content

[0057] After measuring the moisture value with the probe every day, apply 100 mg / mL of prickle fruit polysaccharide and 40-50 mg of positive drug for administration. 4 hours after administration, measure the moisture value again. Figure 4 As shown, the epidermal moisture value of the Primarina utilis group was significantly lower than that of the model group (P<0.05), and had no significant difference with the calcipotriol group (P>0.05).

[0058] (5) ELISA was used to detect the contents of ceramide (Cer), cholesterol (CHOL), IL-17A, IL-23, IL-6, TGF-β, and IL-1β.

[0059] The contents of epidermal ceramide and free fatty acids in the Prinus utilis group were significantly higher than those in the model group (P<0.05), but had no significant difference with the calcipotriol group (P>0.05). Figures 5 to 11 As shown in the figure, the cytokines IL-17A, IL-23, IL-6, IL-1β, and TNF-β were significantly lower than those in the model group (P<0.05), and there was no significant difference between the calcipotriol group and the model group (P>0.05).

[0060] (6) Western blot detection of the expression levels of epidermal barrier-related proteins Filaggrin, Loricrin, and Involucrin

[0061] like Figures 12-14 As shown in the results, the contents of Filaggrin, Loricrin and Involucrin in the epidermis of the Prinus utilis group were significantly higher than those in the model group (P<0.05), but had no significant difference with the calcipotriol group (P>0.05).

[0062] In summary, Prinus utilis polysaccharide can effectively improve psoriasis lesions, and its mechanism is related to its ability to reduce the expression of psoriasis cytokines IL-17A, IL-23, IL-6, IL-1β, and TNF-β, increase the expression of epidermal barrier-related proteins Filaggrin, Loricrin, and Involucrin, and increase intercellular lipid ceramide and free fatty acids.

[0063] The experiment showed that the polysaccharide from Principal Insect Fruit can significantly improve the psoriasis skin lesions. Compared with the model group, the PASI score of mice in the Principal Insect Fruit Polysaccharide group was significantly reduced, indicating its effectiveness in reducing scaling, erythema and infiltration thickening.

[0064] The experimental results show that the polysaccharide from the fruit of black thorn can effectively reduce the transepidermal water loss of the skin and increase the moisture content of the skin, which indicates that it has a positive effect on enhancing the skin barrier function.

[0065] Prinus utilis polysaccharide shows good anti-inflammatory effect by reducing the expression of inflammatory cytokines (such as IL-17A, IL-23, IL-6, IL-1β, TNF-β). The reduction of these cytokines helps to alleviate the inflammatory response caused by psoriasis.

[0066] The present invention increases the expression of epidermal barrier-related proteins (such as Filaggrin, Loricrin, Involucrin), which are essential for maintaining the structure and function of the skin and help improve the skin health of psoriasis patients.

Claims

1. An application of Prinus utilis polysaccharide, characterized in that: The prickle fruit polysaccharide is used to treat and improve psoriasis skin lesions; the prickle fruit polysaccharide is significantly effective in reducing scales, erythema and infiltration and thickening, effectively reduces transepidermal water loss, increases the moisture content of the skin, plays a positive role in enhancing the skin barrier function, significantly reduces the expression of inflammatory cytokines, has a good anti-inflammatory effect, increases the expression of epidermal barrier-related proteins, and helps to improve the skin health of patients with psoriasis.

2. The method for extracting polysaccharides from Principal Utility Fruit according to claim 1, characterized in that: The steps include: The residue after oil is squeezed from the kernel of the prickle fruit is extracted 2-3 times with 95% ethanol at 80-95°C; the ethanol extract is cooled to room temperature and filtered, and the residue is extracted 2-3 times with deionized water at 85-95°C; the extract solution is concentrated and combined under reduced pressure, and then alcohol precipitation is performed with 90%-95% ethanol at 2-6°C; the protein in the precipitate is removed by Sevage method, and the supernatant is collected by centrifugation; the supernatant is freeze-dried to obtain the prickle fruit polysaccharide.

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