CN112107564A - Application of schizandrol A in preparing medicine for preventing and treating ulcerative colitis - Google Patents

Abstract

The invention relates to the field of medicine and pharmacology, and particularly relates to medical application of schisandrin. The schisandrin can be used for preventing and treating ulcerative colitis.

Description

Application of schizandrol A in preparing medicine for preventing and treating ulcerative colitis

Technical Field

The invention belongs to the field of medicine. In particular to application of schisandrin in preparing a medicine for preventing and treating enteritis.

Background

Ulcerative Colitis (UC) is an inflammatory disease of the intestinal tract with an unknown etiology and a high recurrence rate, and constitutes Inflammatory Bowel Disease (IBD) together with Crohn's Disease (CD). UC can occur in all age groups and is difficult to cure, and the incidence rate tends to increase remarkably in China, so that UC is one of modern difficult diseases. Chronic nonspecific inflammation mainly affecting colorectal mucosa due to pathological changes is mainly abdominal pain, diarrhea, mucous bloody stool and tenesmus, is delayed in course of disease, is easy to recur, and has the possibility of canceration. At present, the treatment of UC is mainly based on western medicines, including steroids and non-steroids, such as glucocorticoids, aminosalicylates, antibiotics, immunosuppressants and the like. However, the long-term use of these drugs has large side effects and the drugs are likely to rebound after the drug is stopped, for example, clinical SASP can cause hepatotoxicity. Because Chinese medicine resources are rich, the new application of screening active new compounds or natural compounds with known structures from natural plants becomes one of the main approaches for developing new medicines. Therefore, it is an important subject in the art to find natural drugs for treating ulcerative colitis from natural plants.

Schisandrin A is a lignan compound separated from Schisandra chinensis of Magnoliaceae, and has antiinflammatory, antioxidant, liver protecting, tranquilizing, hypnotic, and blood sugar lowering effects.

Disclosure of Invention

The invention aims to provide a new medical application of schisandrin.

Specifically, the invention provides an application of schizandrol A in preparing a medicine for preventing and treating ulcerative colitis.

In a preferred embodiment, the schisandrin is used as the only active ingredient in the preparation of the medicament for preventing and treating ulcerative colitis.

The details of various aspects of the invention are set forth in subsequent sections. The features, objects, and advantages of the invention will be apparent from the description and from the claims.

Drawings

FIG. 1 shows the effect of schizandrol A on the body weight of mice with ulcerative colitis;

FIG. 2 shows the effect of schizandrol A on the incidence of bloody stool in mice with ulcerative colitis;

FIG. 3 shows the effect of schizandrin A on the length of the colon of mice with ulcerative colitis, FIG. 3A showing the overall appearance of the colon of mice, FIG. 3B showing the length of the colon of mice;

FIG. 4 shows the effect of schizandrin A on colon histopathology in ulcerative colitis mice, FIG. 4A mouse colon HE staining, FIG. 4B histopathology score;

(in the above-mentioned figure: a represents a normal control group, b represents a model control group, c represents a SASP group, and d represents a schizandrol A group)

Detailed Description

The invention arose in part from the unexpected discovery that: the schisandrin can obviously improve the symptoms of weight loss, bloody stool, colon shortening, histopathological injury and the like of experimental ulcerative colitis mice. Therefore, the schisandrin can be used for preparing the medicine for preventing and treating enteritis, in particular ulcerative colitis.

Furthermore, the invention provides the application of the schisandrin in preparing the medicine for preventing and treating enteritis.

The molecular formula of the schisandrin A is as follows: c₂₄H₃₂O₇The molecular weight is: 432.513, having the formula:

the schisandrin of the present invention can be obtained commercially from Chengdou method Biotechnology Co., Ltd, Chengdou mart Biotechnology Co., Ltd, Shanghai source leaf Biotechnology Co., Ltd, etc. The purity of the product meets the pharmaceutical standard. The purity of the schizandrol A is more than or equal to 97 percent optimally.

The schisandrin of the invention can be used alone or in the form of a pharmaceutical composition. The pharmaceutical composition comprises the schisandrin A as an active ingredient and a pharmaceutically acceptable carrier. Preferably, the pharmaceutical composition of the present invention contains 0.1 to 99.9% by weight of the schizandrol a of the present invention as an active ingredient. The pharmaceutical carrier does not damage the pharmaceutical activity of the schisandrin A, and the effective dosage of the schisandrin A is nontoxic to human body when the pharmaceutical carrier acts.

Such pharmaceutically acceptable carriers include, but are not limited to: lecithin, aluminum stearate, alumina, ion exchange materials, self-emulsifying drug delivery systems, tweens or other surfactants, serum proteins, buffer substances such as phosphates, glycine, sorbic acid, water, salts, electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, magnesium silicate, mixtures of saturated fatty acid partial glycerides, and the like.

Other conventional pharmaceutical adjuvants such as binder (e.g. microcrystalline cellulose), filler (e.g. starch, glucose, anhydrous lactose and lactose beads), disintegrant (e.g. crosslinked PVP, croscarmellose sodium, low-substituted hydroxypropylcellulose), lubricant (e.g. magnesium stearate), and absorption enhancer, adsorption carrier, flavoring agent, sweetening agent, excipient, diluent, wetting agent, etc.

The schizandrol A and the pharmaceutical composition thereof of the present invention can be prepared according to conventional methods in the art and can be administered by gastrointestinal or parenteral or topical skin routes. The oral preparation comprises capsule, tablet, oral liquid, granule, pill, powder, pellet, and unguent; parenteral preparations include injections and the like; topical skin preparations include ointments, plasters, rubber plasters, pastes, liniments, lotions, plastics, iontophoresis agents, and the like. Topical dermal formulations are preferred.

The administration route of the schisandrin and the pharmaceutical composition thereof can be oral administration, sublingual administration, transdermal administration, intramuscular administration, subcutaneous administration, skin mucosa administration and the like.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers. All percentages, ratios, proportions, or parts are by weight unless otherwise specified.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

The features mentioned above with reference to the invention, or the features mentioned with reference to the embodiments, can be combined arbitrarily. All the features disclosed in this specification may be combined in any combination and each feature disclosed in this specification may be replaced by alternative features serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the features disclosed are merely generic examples of equivalent or similar features.

The first embodiment is as follows:

1. experimental Material

1.1 medicinal materials

Schisandrin (CAS: 7432-28-2, molecular weight 432.513, HPLC purity not less than 98%, Chengdu Purui method science and technology development Co., Ltd.); dextran sulfate sodium (DSS, CAS: 9011-18-1, MW 36-50kDa, manufactured by MP Biomedicals, USA); sulfasalazine (SASP, CAS: 599-79-1, molecular weight 398.39, HPLC purity 98% or more, manufactured by Sigma-Aldrich Co., USA).

1.2 Experimental animals

C57BL/6J female mice weighing 20 ± 2g (8 weeks), provided by the experimental animals center of the university of medicine, shanghai, animal certification No.: SYXK (Shanghai) 2018-0032. Placing in conventional breeding environment, and freely taking food and drinking water.

2. Experimental methods

2.1 establishment of ulcerative colitis model

Female mice (20 + -2 g) of C57BL/6J, which were of uniform body weight, were selected and given free access to food and water at the beginning of the study using the International general ulcerative colitis modeling method (Gastroenterology 2002,123: 256-70; PLoS One 2012,7: e36075), and water was changed to 4% (w/v) DSS for 8 days after the initial acclimation phase to induce ulcerative colitis.

2.2 methods of grouping and administering drugs

Normal control group: 10, and normal diet.

Model control group: 10, free drinking of 4% DSS solution was given for 8 days.

SASP group: 10 of these, 4% DSS solution was given simultaneously with gastric gavage of SASP (350mg/kg) for 8 consecutive days.

Schizandrol group A: 10 patients were given 4% DSS solution simultaneously with a gavage of schizandrol A (50mg/kg) for 8 consecutive days.

During the experiment, dosage formulations were prepared fresh daily, dissolved in 0.5% sodium carboxymethyl cellulose and used within 1 hour of preparation.

2.3 evaluation of enteritis

Diarrhea, bloody stool, histopathological analysis, etc. were performed according to the methods described in the earlier published papers (Am J Physiol gastroenterest Liver Physiol,2015,309(7): G517-G527). Body weight changes, diarrhea and bloody stool symptoms were recorded daily during the experiment. After the experiment was completed, the mice were sacrificed by anesthesia and cervical dislocation, the abdominal cavity was opened, the colon was removed, and the length of the colon from the cecum to the rectum was measured. Taking the tail colon as a tissue section, carrying out H & E staining, and carrying out pathological scoring on the H & E stained colon specimen: (1) exudation scoring criteria for inflammatory cells: minute quantity of inflammatory cells exist in the 0 minute-mucous membrane inherent layer, more inflammatory cells exist in the 1 minute-mucous membrane inherent layer or the inflammatory cells in the mucous membrane inherent layer are increased, 2 minute-inflammatory cells are diffused to the submucosa, and 3 minute-whole layer has inflammatory cell exudation; (2) tissue damage scoring criteria: 0 point-no mucosal damage, 1 point-discontinuous mucosal epithelial damage, 2 points-superficial mucosal erosion, and 3 points-extensive mucosal damage and deep extension to intestinal wall. The exudation score and tissue damage score of inflammatory cells were added and a histopathological score (1-6 points) was calculated.

2.4 detection of IL-6 content in colonic tissue by ELISA

After the experiment was completed, the mice were sacrificed by anaesthesia and cervical dislocation, the abdominal cavity was opened, the colon was removed, weighed and homogenized. The homogenate was centrifuged at 2000 rpm for 10 minutes at 4 ℃. mu.L of the supernatant was used to determine the IL-6 content using an ELISA kit (R & D systems, Minneapolis, MN, USA), the details of which were referred to the kit instructions. Protein content was determined by BCA method. The results are expressed as pg/mg protein.

2.5 statistical analysis

All experimental data were repeated 3 times, the results were expressed as mean ± standard deviation, and One-way analysis of variance (One-way ANOVA) and LSD test were performed on the experimental data using SPSS 16.0 statistical software, with P <0.05 being statistically significantly different.

3. Results

3.1 therapeutic Effect of Schizandrol A on ulcerative colitis

3.1.1 Effect on body weight in mice with ulcerative colitis

FIG. 1 shows the effect of schizandrol A on the body weight of mice with ulcerative colitis. The body weight of the normal control group mice changed steadily throughout the experiment, with a 0.9% increase in body weight (22.0. + -0.2 g) at day 8; the body weight of the model group mice continuously decreased, and the weight loss rate on day 8 (16.7. + -. 0.2g) was 24.2%; the weight loss rates of mice in the SASP group (17.9 +/-0.3 g) and the schizandrol A group (18.9 +/-0.4 g) are 19.2% and 14.1%, respectively, and are significantly different compared with the model group (P <0.01 and P < 0.001). The SASP (350mg/kg) and the schizandrol A (50mg/kg) can obviously improve the weight loss symptom of the mice with the ulcerative colitis caused by the DSS.

3.1.2 Effect on the incidence of bloody stools in mice with ulcerative colitis

FIG. 2 shows the effect of schizandrol A on the incidence of bloody stool in mice with ulcerative colitis. Throughout the experiment, normal control mice developed no blood. Starting on day 4, 3 groups of mice other than the normal control group began to develop bloody stool symptoms, with the model group mice being the most severe. The incidence of bloody stools on day 8 of the model group was 91.3%; the incidence of bloody stools on day 7 of the SASP group was 60.2%; the schisandrin A group had continuous hematochezia until day 8, with a hematochezia incidence of 55.0%. Compared with the model group, the degrees of diarrhea and bloody stool of the schisandrin A and SASP groups are obviously reduced, and both have obvious difference (P <0.001), which shows that SASP (350mg/kg) and schisandrin A (50mg/kg) can obviously improve the bloody stool symptoms of mice with ulcerative colitis caused by DSS.

3.1.3 Effect on Colon Length in mice with ulcerative colitis

FIG. 3 shows the effect of schizandrin A on colon length in mice with ulcerative colitis. Compared with the normal control group (9.6 +/-0.6 cm), the colon of the model group (4.6 +/-0.3 cm) mice is swollen and bleedings (figure 3A), and is obviously shortened, and the shortening rate is 52.3% (figure 3B); the colons of the mice in the SASP group (6.2 ± 0.2cm) and schizandrin (7.0 ± 0.3cm) were slightly swollen and bleeding (fig. 3A), with shortening rates of 35.4% and 27.1%, respectively (fig. 3B), and were significantly different from the model group (P <0.001 and P < 0.05). The SASP (350mg/kg) and the schizandrol A (50mg/kg) can obviously improve the colon shortening symptom of the mice with the ulcerative colitis caused by the DSS.

3.1.4 Effect on Colon histopathology in mice with ulcerative colitis

FIG. 4 shows the repairing effect of schizandrol A on colon tissue of mouse with ulcerative colitis. Colon tissues of the model group mice are typical inflammatory mucosa manifestations, and can be seen in large-amount inflammatory cell infiltration of mucosa, submucosa and even muscular layer, tissue structure disorder and fracture, submucosa bleeding, edema and telangiectasia; the colon tissue of mice in the SASP (350mg/kg) and schizandrol A (50mg/kg) groups showed reduced lesions, the ulcer healed, and the submucosal bleeding was reduced (FIG. 4A).

Normal control mice were scored as 0 on a 1-6 point scale representative of inflammatory tissue injury; the score of the model group mice (5.8 +/-0.2 points) is obviously increased; mice scored less for both the SASP (score 5 ± 0.1) and schizandrol (score 3.8 ± 0.4) groups and were significantly different compared to the model group (P <0.05 and P <0.01) (fig. 4B).

The colon histopathological analysis result shows that the SASP and the schizandrol A can obviously improve the pathological injury of the colon mucosa tissue and inflammatory cell infiltration symptoms of the mice with the ulcerative colitis caused by the DSS.

3.1.5 Effect on colonic tissue IL-6 content in ulcerative colitis mice

The colon specimen was weighed and homogenized, and the IL-6 content of the supernatant of the homogenate was measured by ELISA kit, the results are shown in Table 1.

TABLE 1 Effect of Schizandrin A on IL-6 content in colon tissue of mice with ulcerative colitis

Table 1 shows the effect of schizandrin A on IL-6 content in colon tissue of mice with ulcerative colitis. Compared with the IL-6 content (25.3 +/-3.0) of the normal control group, the IL-6 content (243.4 +/-16.2) of the colon tissue of the mouse of the model group^###) Significant increase (P)<0.001); intragastric administration of SASP (150.3 + -11.1)^***) Or schizandrol A (100.6 + -9.8)^***) Thereafter, the IL-6 content of the colon tissue of the mouse was decreased, and the difference was significant (P) compared with the model group<0.001). The SASP and the schizandrol A both can obviously reduce the tissue content of IL-6 in inflammatory reaction caused by DSS.

The research results show that the schisandrin can obviously improve the symptoms of the ulcerative colitis mouse such as weight loss, bloody stool, colon shortening, colonic mucosa injury and the like.

The various aspects of the invention are addressed above. It should be understood, however, that equivalent changes and modifications may be made thereto by those skilled in the art without departing from the spirit of the present invention, and that such changes and modifications are intended to be covered by the appended claims.

Claims (2)

1. Application of schisandrin in preparing medicine for preventing and treating ulcerative colitis is provided.

2. The use as claimed in claim 1, wherein schizandrol A is used as the sole active ingredient in the preparation of a medicament for the prevention and treatment of ulcerative colitis.

Images