阅读说明：本技术 柴胡皂苷a在制备防治银屑病的药物中的应用 (Application of saikosaponin a in preparing medicine for preventing and treating psoriasis ) 是由 郑焱 刘萌 邵永平 于 2019-11-11 设计创作，主要内容包括：本发明提供了涉及柴胡皂苷a在制备防治银屑病的药物中的应用,属于生物医药技术领域,所述药物以柴胡皂苷a为活性成分,以生理盐水为溶剂溶解柴胡皂苷a获得。采用本发明所述药物适当的剂量灌胃治疗咪喹莫特银屑病模型小鼠,能够抑制小鼠皮损的炎症反应,改善鳞屑和皮损厚度,能够抑制人表皮角质形成细胞HEKa细胞系的过度增殖；柴胡皂苷a通过诱导细胞线粒体跨膜电位Δψ降低和氧自由基ROS产生来减缓银屑病的进展以及促进皮损恢复正常。相比其它治疗银屑病药物,作用机制更加明确；安全性更好。(The invention provides application of saikosaponin a in preparation of a medicine for preventing and treating psoriasis, and belongs to the technical field of biological medicines. By adopting the medicine disclosed by the invention to treat the imiquimod psoriasis model mouse through intragastric administration at a proper dose, the inflammatory reaction of the mouse skin damage can be inhibited, the scale and skin damage thickness can be improved, and the excessive proliferation of the human epidermal keratinocyte HEKa cell line can be inhibited; saikosaponin a slows the progression of psoriasis and promotes the return of skin lesions to normal by inducing a decrease in the cellular mitochondrial transmembrane potential Δ ψ and the production of oxygen free radical ROS. Compared with other medicines for treating psoriasis, the medicine has more definite action mechanism; the safety is better.)

1. The application of saikosaponin a in preparing the medicine for preventing and treating psoriasis is characterized in that the medicine is obtained by taking saikosaponin a as an active ingredient and dissolving the saikosaponin a by taking normal saline as a solvent.

2. The use of claim 1, wherein the concentration of saikosaponin a in the medicament is 10-20 mg/ml.

3. The use according to claim 2, wherein the concentration of saikosaponin a in the medicament is 12-18 mg/ml.

4. The use according to claim 1, wherein the physiological saline is a NaCl solution with a mass concentration of 0.9%.

5. Application of saikosaponin a in preparing medicine for inhibiting excessive proliferation of human epidermal keratinocyte is provided.

6. Application of saikosaponin a in preparing medicine for reducing expression of inflammatory factor mRNA in psoriasis mouse model is provided.

7. The use of claim 6, wherein said inflammatory factors comprise TNF-a, IL-17A, IL-8 and IL-1 β.

8. Application of saikosaponin a in preparing medicine for inducing reduction of mitochondrial transmembrane potential delta psi in cells is provided.

9. Application of saikosaponin a in preparing medicine for inducing the generation of oxygen free radical ROS in cells is provided.

Technical Field

The invention belongs to the technical field of biological medicines, and particularly relates to application of saikosaponin a in preparation of a medicine for preventing and treating psoriasis.

Background

Psoriasis is common immune-mediated chronic recurrent inflammatory skin diseases and is characterized by epidermal hyperplasia, immune cell infiltration, skin angiogenesis increase and local upregulation of various inflammatory mediators, typical skin lesions of psoriasis are represented by squamous erythema or plaques, repeated attacks and long-term courses of the psoriasis seriously affect the body, psychology, spirit and the like of patients, the organization and pathology of psoriasis are mainly represented by hyperkeratosis and parakeratosis, neutrophilic granulocyte aggregation is often seen in parakeratosis areas, the area is called Munro micro-abscess, epidermal process is extended, dermal papilla is extended to be in a shape, the upper spinous layer is thinned, capillary vessels in papilla are circuitously dilated and congested, peritubular inflammatory cell infiltration is achieved, the psoriasis is mainly treated by external drugs in a mild way, is treated by a moderate and severe available system, can be treated by properly selecting a targeted biological agent for patients with poor treatment effect of the traditional systemic drugs, and the like, but the psoriasis still faces difficulty and is the key point of research.

Bupleurum root, belonging to the dried root of Bupleurum scorzonerifolium or Bupleurum scorzonerifolium of Umbelliferae, has various pharmacological actions, such as immunoregulation, anti-inflammation and liver protection, anti-tumor and anti-cell adhesion. The main components of bupleurum root can be divided into saikosaponin a, b, c and d according to different chemical structures, and the saikosaponin a and the saikosaponin d are taken as main components, have various biological activities, and are important indexes for evaluating the quality of bupleurum root.

Saikosaponin a belongs to penta-triterpenoid oleanane type derivative, and its chemical structural formula is C₄₂H₆₈O₁₃Molecular weight is 780.98, and its chemical structural formula is shown in figure 1. Saikosaponin a has antiinflammatory, immunity regulating, and antiviral effects.

At present, the mechanism and the effect of the saikosaponin a for preventing and treating the psoriasis are not clear.

Disclosure of Invention

In view of the above, the invention aims to provide application of saikosaponin a in preparing a medicine for preventing and treating psoriasis.

In order to achieve the above purpose, the invention provides the following technical scheme:

the invention provides application of saikosaponin a in preparing a medicine for preventing and treating psoriasis, and the medicine

The saikosaponin a is obtained by dissolving saikosaponin a with normal saline as solvent.

Preferably, the concentration of the saikosaponin a in the medicine is 10-20 mg/ml.

Preferably, the concentration of the saikosaponin a in the medicine is 12-18 mg/ml.

Preferably, the physiological saline is a NaCl solution with a mass concentration of 0.9%.

The invention provides application of saikosaponin a in preparing a medicament for inhibiting excessive proliferation of human epidermal keratinocytes.

The invention provides application of saikosaponin a in preparing a medicament for reducing expression of inflammatory factor mRNA in a psoriasis mouse model.

Preferably, the inflammatory factors include TNF-a, IL-17A, IL-8 and IL-1 β.

The invention provides application of saikosaponin a in preparing a medicament for inducing reduction of mitochondrial transmembrane potential delta psi in cells.

The invention provides application of saikosaponin a in preparation of a medicine for inducing generation of oxygen free radicals ROS in cells.

The invention has the beneficial effects that: the application of the saikosaponin a in preparing the medicine for preventing and treating psoriasis is obtained by dissolving the saikosaponin a with normal saline as a solvent by taking the saikosaponin a as an active ingredient. By adopting the medicine disclosed by the invention to treat the imiquimod psoriasis model mouse through intragastric administration at a proper dose, the inflammatory reaction of the mouse skin damage can be inhibited, and the scales and the skin damage thickness can be improved; the medicine can inhibit hyperproliferation of human epidermal keratinocyte HEKa cell line, thereby inhibiting psoriasis. Saikosaponin a slows the progression of psoriasis and promotes the return of skin lesions to normal by inducing a decrease in the cellular mitochondrial transmembrane potential Δ ψ and the production of oxygen free radical ROS. The saikosaponin a can inhibit psoriasis by inducing ROS generation in a psoriasis pathogenesis channel and reducing the expression level of inflammatory factors, and compared with other psoriasis treatment medicines, the saikosaponin a has a more definite action mechanism; the saikosaponin a and the physiological saline serving as the auxiliary material in the medicine have no toxic or side effect on human bodies and have good safety.

Drawings

FIG. 1 is the molecular structural formula of saikosaponin a;

FIG. 2 is a clinical photograph of different doses of saikosaponin a in mice treated by gavage with imiquimod, wherein the positive control group is 0.5mg/kg/d methotrexate, the negative control group is 0.9% normal saline, and the treatment groups are 37.5mg/kg/d, 75mg/kg/d and 150mg/kg/d saikosaponin a;

FIG. 3 is a PASI score for control and treated imiquimod mice, wherein a is the PASI score for erythema, b is the PASI score for scaling, c is the PASI score for skin thickness, and d is the total PASI score;

FIG. 4 is a histological photograph of skin lesions of control and treated imiquimod mice;

FIG. 5 is a HPSS score for skin lesions in control and treatment groups of imiquimod mice, where a is the score for Epidermis (Epidermis), b is the score for vascular (vascular) changes, and c is the score for Infiltration (Infiltration);

FIG. 6 shows the total RNA extracted from the skin lesions of the control and treated imiquimod mice, and the relative expression change of psoriasis-related inflammatory factors detected by real-timePCR, wherein a is interleukin-17, b is tumor necrosis factor (TNF-a), and c is interleukin-1 β.

FIG. 7 shows that after HEKa cells are stimulated by saikoside a with different concentrations for 24h, 48h and 72h, the MTT method is used to detect the effect on HEKa cell proliferation;

FIG. 8 shows the change of mitochondrial membrane potential Δ ψ of each group after different concentrations of saikoside a stimulate HEKa cells for 24 h;

FIG. 9 shows the change of ROS content in each group after different concentrations of saikoside a stimulate HEKa cells for 24 h.

Detailed Description

The invention provides application of saikosaponin a in preparing a medicine for preventing and treating psoriasis, wherein the medicine is obtained by dissolving saikosaponin a as an active ingredient and normal saline as a solvent.

In the invention, the concentration of the saikosaponin a in the medicine is preferably 10-20 mg/ml, more preferably 12-18 mg/ml, and most preferably 15 mg/ml. In the present invention, the physiological saline is preferably a NaCl solution with a mass concentration of 0.9%. The invention has no special limitation on the sources and specifications of the saikosaponin a and the normal saline, and only needs to adopt the conventional commercial products in the field; the physiological saline is preferably obtained by self-preparation.

In the present invention, the preparation method of the drug is preferably as follows: centrifuging the saikosaponin a powder, and dissolving in physiological saline to obtain suspension. The rotation speed of the centrifugation is preferably 1500-2500 rpm, and more preferably 2000 rpm; the centrifugation time is preferably 50-70 s, and more preferably 60 s; the purpose of the centrifugation in the invention is to centrifuge the mural drug to the bottom of the tube and fully dissolve the mural drug.

The invention provides application of saikosaponin a in preparing a medicament for inhibiting excessive proliferation of human epidermal keratinocytes. In the invention, the human epidermal keratinocytes are preferably selected from HEKa cells for testing, and the influence on the proliferation of the HEKa cells is preferably detected by using an MTT method after the human epidermal keratinocytes are stimulated by using saikosaponin a with different concentrations for 24h, 48h and 72 h. The experimental result of the invention shows that the saikosaponin a can inhibit the excessive proliferation of human epidermal keratinocytes.

The invention provides application of saikosaponin a in preparing a medicine for reducing expression of mRNA of an inflammatory factor in a psoriasis mouse model, the inflammatory factor preferably comprises TNF-a, IL-17A, IL-8 and IL-1 β, and the expression of the mRNA of the inflammatory factor is preferably detected by real-time PCR.

The invention provides application of saikosaponin a in preparing a medicament for inducing reduction of mitochondrial transmembrane potential delta psi in cells. According to the invention, after different concentrations of saikosaponin a are adopted to stimulate the aHEKa cells for 24h, 48h and 72h, the mitochondrial transmembrane potential delta psi is detected, and the mitochondrial transmembrane potential delta psi is obviously reduced.

The invention also provides application of the saikosaponin a in preparing a medicament for inducing the generation of oxygen free radicals ROS in cells. According to the invention, after the saikosaponin a with different concentrations is adopted to stimulate the aHEKa cells for 24h, 48h and 72h, the oxygen free radical ROS in the cells is detected, and the level of the oxygen free radical ROS in the cells is obviously increased.

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