Application of kadsura root-bark extract in preventing and treating inflammatory bowel disease

Abstract

The invention provides application of a Chinese redbud bark extract in preventing and treating inflammatory bowel diseases. Relates to the technical field of medicine. The kadsura root-bark extract is obtained by performing heating reflux twice and vacuum concentration drying on dried kadsura root-bark. The kadsura root-bark extract can prevent colon shortening, inhibit the expression level of myeloperoxidase (MPO) and improve the pathological structure of colon tissue so as to prevent and treat the inflammatory bowel disease, especially colitis, and is applied to preparation of drugs, food or health care products for preventing and treating the inflammatory bowel disease.

Description

Technical Field

[0001] The invention relates to the technical field of medicines, and in particular to an application of a Bauhinia bark extract for preventing and treating inflammatory bowel disease. Background Art

[0002] Inflammatory Bowel Disease (IBD) is a type of disease characterized by chronic intestinal inflammation, mainly including ulcerative colitis (UC) and Crohn's disease (CD). Its specific cause is not yet fully understood, but it is generally believed to be related to genetic factors, immune system disorders, environmental factors and changes in intestinal flora. In recent years, the incidence of inflammatory bowel disease has been increasing worldwide.

[0003] Inflammatory bowel disease is characterized by persistent inflammation of the intestine, which may lead to severe gastrointestinal symptoms and systemic complications. Ulcerative colitis mainly affects the colon and rectum, and is often manifested by abdominal pain, diarrhea (usually accompanied by blood and mucus), tenesmus, and weight loss; Crohn's disease can affect the entire digestive tract, and clinical symptoms include right lower abdominal pain, diarrhea, gastrointestinal obstruction, fistula formation, weight loss, and malnutrition. Both diseases can present with extraintestinal symptoms such as arthritis, skin lesions (such as erythema nodosum), eye inflammation, and liver disease.

[0004] In the study of animal models, commonly used inflammatory bowel disease models include chemically induced models and transgenic models. Chemically induced models such as dimethyl sulfoxide (DSS) and trifluorochlorothiol (TNBS) are used to induce colitis in mice or rats, simulating the pathological characteristics of ulcerative colitis and Crohn's disease. Transgenic models, such as IL-10-deficient mice, are prone to develop colitis due to their defective immune system and are widely used to study immune-mediated enteritis.

[0005] The diagnosis and monitoring of IBD rely on a variety of disease markers, including serum markers, fecal markers, and histological markers. Common serum markers include myeloperoxidase (MPO) and C-reactive protein (CRP), which reflect the inflammatory state in the body; fecal markers such as fecal calprotectin and lactoferrin can reflect the level of intestinal inflammation; in addition, histological examination can also provide a basis for the diagnosis of IBD by observing the morphological changes of intestinal tissue and the degree of infiltration of inflammatory cells.

[0006] At present, the drug treatment of IBD mainly includes salicylic acid preparations, glucocorticoids, immunosuppressants, biological agents, etc. Although the existing treatment methods can alleviate the condition to a certain extent, there are still problems such as poor treatment effect, prolonged ulcers, large side effects, and easy recurrence, which have a serious impact on the quality of life of patients. Therefore, there is an urgent need to develop more safe and efficient therapeutic drugs. Therefore, finding safe and effective inflammatory bowel disease (IBD) therapeutic drugs from natural products of traditional Chinese medicine has become a new research direction.

[0007] Bauhinia bark is the bark of Cercis chinensis Bunge, a plant of the Leguminosae family. The earliest record of its nature and flavor can be found in the Song Dynasty's "Kaibao Materia Medica", which records that Bauhinia wood is bitter, flat, non-toxic, and has the effect of "breaking old blood and relieving stranguria". The Ming Dynasty's "Compendium of Materia Medica" further supplemented the efficacy of Bauhinia bark on the basis of previous research, adding "diuresis and detoxification" and other effects. In clinical prescription applications, the efficacy and main indications of Bauhinia bark have been further expanded. The Song Dynasty's "Ren Zhai Zhi Zhi" records that Bauhinia bark can be used to treat various swellings, ulcers, and joint pains; the Ming Dynasty's "Xianchuan Surgical Secret Prescriptions" uses Chonghe Xian Gao to treat primary carbuncles with unclear cold and heat, back bleeding, and other symptoms; the Qing Dynasty's "Shangke Huizan" uses it to treat eye swelling and pain, dog bites, and insect poisons.

[0008] Bauhinia bark has the effects of promoting blood circulation, relieving stranguria, reducing swelling and detoxifying, and is mainly used to treat rheumatic arthralgia, abdominal pain caused by stasis, irregular menstruation, painful urination, scabies, carbuncle, swelling, injuries from falls, insect and snake bites, etc. In clinical practice, Bauhinia bark is often used for external application to reduce swelling and relieve pain. For example, Bauhinia pain relief patch with Bauhinia bark, rhubarb, catechu, borneol, and menthol as main ingredients can be used to treat acute ankle sprains, which can effectively reduce pain in the affected area, eliminate swelling, and improve ankle function.

[0009] Studies have shown that the ethyl acetate extract of Bauhinia bark can be used to relieve acute inflammation and chronic inflammation of granulomas, and has good anti-inflammatory and analgesic activity. In addition, there are studies that extracts obtained by drying after reflux extraction with 75% ethanol and 95% ethanol twice have a significant inhibitory effect on primary and secondary foot lesions in rat adjuvant arthritis models, and are potential drugs for the treatment of rheumatoid arthritis. However, there are no literature reports on the use of its extracts in the prevention and treatment of inflammatory bowel disease and other gastrointestinal inflammatory diseases at home and abroad. Summary of the invention

[0010] In view of the technical problems existing in the prior art, the present invention aims to provide an application of a Bauhinia bark extract in preventing and treating inflammatory bowel disease.

[0011] Preferably, the Bauhinia bark extract is used to prepare a medicament for preventing and treating inflammatory bowel disease.

[0012] Preferably, the inflammatory bowel disease comprises colitis.

[0013] Preferably, the drug is prepared into a pharmaceutically acceptable dosage form with the extract as the active ingredient, alone or in combination with other drugs, and with the addition of pharmaceutically acceptable excipients, auxiliary ingredients or carriers.

[0014] Preferably, the dosage form is a hard capsule, a soft capsule, a powder, a granule, a tablet, a pill, a honey paste, an oral solution, a suppository, an alcoholic solution or an injection.

[0015] Preferably, the preparation method of the Bauhinia bark extract comprises: The dried Bauhinia bark was extracted twice with an organic solvent under heating and reflux, and the obtained extract was vacuum concentrated and dried to constant weight to obtain a brown paste extract.

[0016] Preferably, the organic solvent includes one or more of 75% ethanol, ethyl acetate, petroleum ether, propanol, glycerol or methanol.

[0017] Preferably, the mass ratio of the organic solvent to the dried Bauhinia bark is 4-5:1.

[0018] Preferably, the heating reflux is performed twice, the first temperature is 80° C., the reflux time is 1.5-2.5 h, and the second temperature is 80° C., the reflux time is 1-2 h.

[0019] Preferably, the vacuum drying temperature is 70-90°C.

[0020] Beneficial effects of the present invention: The present invention provides an application of a Bauhinia bark extract in preventing and treating inflammatory bowel disease. The Bauhinia bark extract can prevent colon shortening, inhibit the expression level of myeloperoxidase (MPO), improve the pathological structure of colon tissue, and thus prevent and treat inflammatory bowel disease, especially colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 This is the weight change trend of mice within 12 days; Figure 2 This is the changing trend of the disease activity index (DAI) of mice within 12 days; Figure 3 Photographs of mouse colon measurements; Figure 4 Comparison of mouse colon length; Figure 5 This is the H&E staining picture of mouse colon tissue; Figure 6 is the MPO expression level. DETAILED DESCRIPTION

[0022] The invention provides an application of a Bauhinia bark extract in preventing and treating inflammatory bowel disease. The Bauhinia bark extract is used in preparing a medicine for preventing and treating inflammatory bowel disease.

[0023] Bauhinia bark extract can prevent colon shortening and inhibit the expression level of myeloperoxidase (MPO), thereby preventing and treating inflammatory bowel disease.

[0024] In a preferred embodiment of the present invention, the inflammatory bowel disease includes colitis.

[0025] In a preferred embodiment of the present invention, the drug is prepared into a pharmaceutically acceptable dosage form with the extract as the active ingredient, alone or in combination with other drugs, and with the addition of pharmaceutically acceptable excipients, auxiliary ingredients or carriers.

[0026] In a preferred embodiment of the present invention, the dosage form is a hard capsule, a soft capsule, a powder, a granule, a tablet, a pill, a honey paste, an oral liquid, a suppository, an alcoholic solution or an injection.

[0027] In the present invention, the medicine or health care product can be prepared into various dosage forms known in the art according to actual needs, such as tablets, capsules, pills, etc. in solid preparations, injections, solutions, etc. in liquid preparations, gels in semi-solid preparations, aerosols, sprays, etc. in gas preparations. Alternatively, the food can be prepared into types known in the art, such as biscuits, bread, etc. according to actual needs.

[0028] Those skilled in the art can also understand that when the Bauhinia bark extract of the present invention is used as the main active ingredient to prepare medicines, foods, and health products for preventing or treating inflammatory bowel inflammation, conventional adjuvants such as leveling agents, fillers, wetting agents, disintegrants, solubilizers, lubricants, etc. can be added according to actual needs.

[0029] In a preferred embodiment of the present invention, the preparation method of the Bauhinia bark extract comprises: The dried Bauhinia bark was extracted twice with an organic solvent under heating and reflux, and the obtained extract was vacuum concentrated and dried to constant weight to obtain a brown paste extract.

[0030] In a preferred embodiment of the present invention, the organic solvent includes one or more of 75% ethanol, ethyl acetate, petroleum ether, propanol, glycerol or methanol.

[0031] In a preferred embodiment of the present invention, the mass ratio of the organic solvent to the dried Bauhinia bark is 4-5:1.

[0032] In a preferred embodiment of the present invention, the heating reflux is performed twice, the first temperature is 80° C., the reflux time is 1.5-2.5 h, and the second temperature is 80° C., the reflux time is 1-2 h.

[0033] In a preferred embodiment of the present invention, the vacuum drying temperature is 70-90°C.

[0034] Example 1 Solvent preparation: Under stirring and heating conditions, 0.5 g of dry sodium carboxymethyl cellulose (TargetMol T19232) was slowly added to 100 ml of double distilled water, and heated and ultrasonicated for 10 minutes to prepare a colorless and clear 0.5% sodium carboxymethyl cellulose solution as a solvent.

[0035] Preparation of Bauhinia bark sample for administration: 400 g of dried Bauhinia bark was placed in a 10L round-bottom flask, and extracted twice with 75% ethanol under heating and reflux, each time using 2L of solvent, the first heating temperature was 80°C, the reflux time was 2 hours, and the second heating temperature was 80°C, the reflux time was 1.5 hours. The obtained extract was vacuum concentrated and vacuum dried at 80°C to constant weight to obtain 62.4 g of brown paste extract, namely Bauhinia bark extract.

[0036] Weigh 9.36 g of Bauhinia bark extract, dissolve it in a solvent with a total volume of 50 ml, and stir thoroughly until a brown uniform suspension is formed, i.e., a Bauhinia bark alcohol extract solution with a concentration of 0.187 g / ml (equivalent to a Bauhinia bark crude drug concentration of 1.2 g / ml).

[0037] Preparation of Sulfasalazine Solution: Weigh 6 g of sulfasalazine and dissolve it in 100 ml of solvent, stir thoroughly, and obtain a 60 mg / ml sulfasalazine solution.

[0038] Preparation of Dextran Sulfate Sodium Salt (DSS) solution: Weigh DSS powder (36,000-50,000 kDa, Yisheng Bio 60316ES76) and dissolve it in sterile drinking water to prepare a 3% solution, which was then sterilized by filtering using a 0.22 μm filter membrane.

[0039] Example 2 Animal grouping and experiments After 5 days of adaptive feeding, the mice were randomly divided into 5 groups: ① healthy control group (Control), ② model group (Model), ③ low-dose Bauhinia bark group (ZJP-L), ④ high-dose Bauhinia bark group (ZJP-H), and ⑤ positive control group (Positive). The low-dose Bauhinia bark group (ZJP-L) and high-dose Bauhinia bark group (ZJP-H) were gavaged with Bauhinia bark alcohol extract solution equivalent to 6g / kg and 12g / kg Bauhinia bark crude drug, respectively, the positive control group (Positive) was gavaged with sulfasalazine solution equivalent to 0.6mg / g sulfasalazine, and the healthy control group (Control) and model group (Model) were gavaged with the same volume of 0.5% sodium carboxymethyl cellulose solution. All groups were routinely fed and had normal drinking water. After 3 days of continuous preventive administration, the healthy control group (Control) was still fed with normal drinking water, and the other groups were allowed to drink 3% DSS solution freely for 7 consecutive days to induce ulcerative colitis. On the 8th day, the water was switched back to normal purified water. After 5 days of continued drug treatment, the mice were killed and their body weights were recorded every day. It was observed that the mice in the model group showed symptoms such as weight loss, thinning feces, and reduced activity, indicating that the DSS-induced colitis model was successfully established.

[0040] (1) Changes in mouse body weight and disease activity index (DAI) Figure 1 The weight change trend of mice within 12 days. Figure 2 The change trend of the disease activity index (DAI) of mice within 12 days is shown in Figure 2. Figure 1 As shown in the figure, compared with the normal group, the body weight of mice in the model group was significantly reduced. After enema administration of Bauhinia bark extract, the weight loss of mice was reduced, and the effect on the weight change of mice was dose-dependent. The effect of the high-dose group (ZJP-H) on the weight change of mice was more significant than that of the low-dose group (ZJP-L).

[0041] like Figure 2 As shown in the figure, the DAI score of mice in the model group was the highest, indicating that their symptoms were more severe than those of mice in other groups. After treatment with Bauhinia bark extract, the DAI score was significantly reduced, indicating that Bauhinia bark can alleviate the symptoms of DSS-induced colitis.

[0042] (2) Comparison of colon length Figure 3 This is a photo of mouse colon measurement. Figure 4 Comparison of mouse colon length.

[0043] like Figure 3 and Figure 4As shown in the figure, after DSS treatment, the colon length was significantly shortened compared with the normal group. In contrast, the colon length of mice in the low-dose Bauhinia bark group (ZJP-L), high-dose Bauhinia bark group (ZJP-H) and positive control group (Positive) was basically the same as that of mice in the normal control group. Therefore, oral administration of Bauhinia bark extract can prevent colon shortening.

[0044] There was no significant difference in the effect of high-dose Bauhinia bark extract on the colon length of colitis mice compared with low-dose.

[0045] (3) H&E staining results H&E staining was used to determine the pathological changes in colon tissues of mice in different groups. Figure 5 As shown in the figure, compared with the normal group, the model group showed severe mucosal ulcers, the basic structure of the mucosa was completely destroyed, the mucosal layer basically disappeared and was replaced by a large amount of granulation tissue, the crypts disappeared, no obvious glandular epithelium was seen, the submucosal layer was obviously loose and edematous, and the inflammatory cell infiltration was obvious. In the ZJP-L group and the ZJP-H group, there were fewer mucosal ulcers in the colon tissue, the basic structure of the mucosa existed, the gland arrangement, density, and morphology were good, the submucosal layer and mucosal muscular layer were roughly normal in morphology, the inflammatory cell infiltration was less, the crypt damage was mild, and there was no significant difference between the groups. It can be seen that the tissue morphology was significantly improved after treatment with Bauhinia bark.

[0046] (4) Determination of myeloperoxidase (MPO) content Experimental materials: Enzyme-linked immunosorbent assay (ELISA) kit (Fine Bio EM0010) MPO is a commonly used inflammatory marker and plays an important role in the occurrence and development of IBD. The distal colon tissue of each group of mice was crushed and centrifuged using a glass homogenizer, and the protein content in the supernatant was determined using the bicinchoninic acid (BCA) method. The expression level of MPO per gram of total protein in colon tissue was then detected using an enzyme-linked immunosorbent assay (ELISA) kit. Figure 6 As shown, the expression level of MPO was significantly inhibited in the Bauhinia bark treatment group.

Claims

1. Application of a Bauhinia bark extract in preventing and treating inflammatory bowel disease, It is characterized in that The Bauhinia bark extract is used for preparing medicines, foods or health products for preventing and treating inflammatory bowel disease.

2. The use according to claim 1, It is characterized in that The inflammatory bowel disease includes ulcerative colitis.

3. The use according to claim 1, It is characterized in that The medicine uses the extract as an active ingredient, alone or in combination with other medicines, and is added with pharmaceutically acceptable excipients, auxiliary ingredients or carriers to prepare a pharmaceutically acceptable dosage form.

4. The use according to claim 3, It is characterized in that The dosage form is hard capsule, soft capsule, powder, granule, tablet, pill, honey paste, oral liquid, suppository, wine or injection.

5. The use according to claim 1, It is characterized in that The preparation method of the Bauhinia bark extract comprises: The dried Bauhinia bark was extracted twice with an organic solvent under heating and reflux, and the obtained extract was vacuum concentrated and dried to constant weight to obtain a brown paste extract.

6. The use according to claim 5, It is characterized in that The organic solvent includes one or more of 75% ethanol, ethyl acetate, petroleum ether, propanol, glycerol or methanol.

7. The use according to claim 5, It is characterized in that The mass ratio of the organic solvent to the dried Bauhinia bark is 4-5:

1.

8. The use according to claim 5, It is characterized in that The heating reflux is carried out twice, the first time at a temperature of 80° C., the reflux time is 1.5 to 2.5 h, and the second time at a temperature of 80° C., the reflux time is 1 to 2 h.

9. The use according to claim 5, It is characterized in that The vacuum drying temperature is 70-90°C.

Images