Application of ethanol extract of wild horsetail and its flavonoids in preparing medicine for preventing or treating ulcerative colitis

Abstract

The invention discloses an application of an ethanol extract of wild horsetail and its flavonoids in preparing a drug for preventing or treating ulcerative colitis. The invention uses sodium dextran sulfate to induce mice to establish an ulcerative colitis mouse model, and uses the ethanol extract of wild horsetail and the flavonoids of wild horsetail extracted from wild horsetail as therapeutic drugs for treating ulcerative colitis, and then observes the effects of the therapeutic drugs on the pharmacological indicators of ulcerative colitis mice. The experimental results show that the ethanol extract of wild horsetail and the flavonoids of wild horsetail can improve the inflammation level in ulcerative colitis mice, inhibit the complement level, significantly reduce the DAI score, reduce the shortening degree of the colon, alleviate the symptoms of congestion and swelling of the colon, and reduce the spleen index. The flavonoids of wild horsetail can also improve colon tissue damage, showing a significant anti-ulcerative colitis effect, and can be used to prepare drugs for preventing and treating ulcerative colitis.

Description

Technical Field

[0001] The invention relates to the technical field of biomedicine, and in particular to application of an ethanol extract of Herba Cynoglossi and a flavonoid fraction thereof in preparing a drug for preventing or treating ulcerative colitis. Background Art

[0002] Ulcerative colitis (UC) is a nonspecific inflammatory disease of unknown cause. The lesions are located in the large intestine and are distributed continuously and diffusely, mostly involving the rectum and sigmoid colon. Its typical clinical course includes bloody stools with mucus, pus, abdominal pain, and diarrhea. The disease lasts for a long time and is prone to recurrence and even canceration, which seriously affects the patient's quality of life. At present, the true cause of UC is still unclear. Multiple factors such as immune regulation dysfunction, genetic factors, smoking, diet, living environment, infection, and mental and psychological factors may promote the occurrence and development of this disease. Clinically, the treatment of UC is mainly based on drugs. ① Aminosalicylic acid preparations, such as sulfasalazine, have a long medication cycle and are prone to dose-related adverse reactions, such as gastrointestinal symptoms, urticaria, dyspnea, joint pain, and hepatotoxicity; ② Glucocorticoids and immunosuppressants have many adverse reactions and cannot control UC recurrence; ③ Anti-tumor necrosis factor α monoclonal antibodies are prone to recurrence and have many adverse reactions; ④ Probiotic therapy has a general effect on improving UC symptoms and is often used in combination with other drugs in clinical practice; ⑤ Enema treatments such as tin powder, gentamicin, 2% procaine, and dexamethasone have poor patient compliance (Li Nan, Research Progress in Ulcerative Colitis, Jilin Medicine, 2014, 35(7):1486-1487.; Ma Qinglin, Du Lidong, Zang Kaihong, Sun Min, Cao Rubing, Ren Yuan, Overview of Research Progress in Ulcerative Colitis, Journal of Medical Forum, 2020, 41(1):175-177. Taku Kobayashi, Britta Siegmund, Catherine Le Berre, Shu Chen Wei, Marc Ferrante, Bo Shen, Charles N. Bernstein, Silvio Danese, Laurent Peyrin-Biroulet & Toshifumi Hibi. Ulcerative colitis. Nature Reviews Disease Primers. 2020, 6: 74.).

[0003] The flavonoid fraction of Eupatorium lindleyanum refers to the flavonoid fraction of Eupatorium lindleyanum containing cyanidin and lutein, which has anti-acute lung injury and anti-hepatitis B virus activity (Chunjun Chu, ShiYao, Jinglei Chen, XiaochenWei, Long Xia, Daofeng Chen, Jian Zhang. Eupatorium lindleyanum DC. flavonoids fraction attenuates lipopolysaccharide-induced acute lung injury inmice. International Immunopharmacology. 2016, 39: 23-33.; Zhang Jian, Chen Daofeng, Ren Huiling, Chu Chunjun, Li Xianlun, Yan Biao, Wu Shuangqing, Application of the flavonoid fraction of Eupatorium lindleyanum in the preparation of anti-hepatitis B virus drugs, ZL201810046069.6).

[0004] At present, there is no report on whether the flavonoids of Yemazhui have an anti-ulcerative colitis effect. The present invention finds that the flavonoids of Yemazhui have an anti-ulcerative colitis effect, so the flavonoids of Yemazhui can be used to prepare drugs related to the prevention and treatment of ulcerative colitis. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide an application of an ethanol extract of Herba Cynoglossi and a flavonoid fraction thereof in preparing a medicine for preventing or treating ulcerative colitis.

[0006] In order to solve the above technical problems, the present invention provides the following technical solutions:

[0007] The first aspect of the present invention provides the use of the flavonoid fraction of YEMAZHUAI in preparing a drug for preventing or treating ulcerative colitis.

[0008] Furthermore, the flavonoid fraction of Herba Cynanchii is preferably a flavonoid fraction of Herba Cynanchii that contains cyanidin and lutein.

[0009] Furthermore, the preparation of the flavonoid fraction of YEMAZHUAI comprises the following steps:

[0010] (1) taking the dried whole herb of wild horsetail, cutting it into 2-3 cm segments, adding 8-15 times the volume of 60% to 95% ethanol aqueous solution, refluxing for 1-3 hours, and recovering the solvent under reduced pressure at 40-60° C. to obtain wild horsetail ethanol extract;

[0011] (2) dissolving the ethanol extract of the wild horsetail prepared in step (1) in water to prepare a 0.1-0.2 g / mL solution, adsorbing it on a weakly polar macroporous adsorption resin, first eluting it with water to remove macromolecular compounds, then eluting it with a volume fraction of 90%-95% ethanol aqueous solution, and recovering the solvent under reduced pressure at 40-60° C. to obtain a macroporous resin fraction;

[0012] (3) The macroporous resin portion prepared in step (2) is adsorbed on an ODS reverse phase silica gel column, eluted with a 50% by volume methanol aqueous solution, then eluted with a 70%-75% by volume methanol aqueous solution, and the solvent is recovered under reduced pressure at 40-60° C. to obtain an extract, and the extract is then adsorbed on a 30-60 mesh polyamide column, first eluted with a 70% by volume ethanol aqueous solution, then eluted with a 95% by volume ethanol aqueous solution, and the solvent is recovered under reduced pressure at 40-60° C. to obtain the wild horsetail flavonoid portion.

[0013] Furthermore, in step (2), the weakly polar macroporous adsorption resin is preferably AB-8, D101, HPD-100, HPD-200A, HPD-200B, HPD-300, HPD-700 or HPD-722 macroporous adsorption resin.

[0014] Furthermore, in step (2), in the step of removing the macromolecular compounds by eluting with water, the volume ratio of water to the weakly polar macroporous adsorption resin is 8-15:1, for example 8:1, 9:1, 10:1, 12:1, 13:1, 14:1, 15:1, etc., including but not limited to the volume ratios listed above.

[0015] Furthermore, in step (2), in the step of eluting with a 90%-95% ethanol aqueous solution by volume, the volume ratio of the 90%-95% ethanol aqueous solution to the weakly polar macroporous adsorption resin is 8-15:1, for example, 8:1, 9:1, 10:1, 12:1, 13:1, 14:1, 15:1, etc., including but not limited to the volume ratios listed above.

[0016] Furthermore, in step (3), the ODS reverse-phase silica gel chromatographic column is a DAISOGEL ODS reverse-phase silica gel chromatographic column, a Cosmsil ODS reverse-phase silica gel chromatographic column or a YMCODS-A reverse-phase silica gel chromatographic column; the filler particle size of the ODS reverse-phase silica gel chromatographic column is 50-70 μm, for example, 50 μm, 55 μm, 60 μm, 65 μm, 70 μm, etc., including but not limited to the particle sizes listed above.

[0017] Furthermore, in step (3), in the step of eluting with a 50% methanol aqueous solution by volume, the volume ratio of the 50% methanol aqueous solution to the ODS reverse phase silica gel chromatography column is preferably 4-6:1.

[0018] Furthermore, in step (3), in the step of eluting with 70%-75% methanol aqueous solution, the volume ratio of the 70%-75% methanol aqueous solution to the ODS reverse phase silica gel chromatography column is preferably 4-6:1.

[0019] Furthermore, in step (3), the mass volume ratio of the extract to the polyamide chromatographic column is 1 mg:20-30 mL, for example 1:25 mg / mL.

[0020] Furthermore, in step (3), in the step of eluting with 70% ethanol aqueous solution by volume, the volume ratio of the 70% ethanol aqueous solution to the polyamide chromatographic column is 3-5:1, for example 4:1.

[0021] Furthermore, in step (3), in the step of eluting with a 95% ethanol aqueous solution by volume, the volume ratio of the 95% ethanol aqueous solution to the polyamide chromatographic column is 3-5:1, for example 4:1.

[0022] Furthermore, in step (3), the flavonoid part of the wild horsetail contains 63%-68% of cyanidins and 24%-29% of luteolin.

[0023] The second aspect of the present invention provides a use of an ethanol extract of Herba Eupatorii in preparing a drug for preventing or treating ulcerative colitis. The ethanol extract of Herba Eupatorii is an extract obtained by treating Herba Eupatorii with a 60%-95% ethanol aqueous solution.

[0024] Furthermore, the preparation of the ethanol extract of the wild horsetail is as follows: take the dried whole herb of the wild horsetail, cut it into 2-3 cm segments, add 8-15 times the volume of a 60%-95% ethanol aqueous solution, reflux extraction for 1-3 hours, and recover the solvent under reduced pressure at 40-60°C to obtain the ethanol extract of the wild horsetail.

[0025] Furthermore, in the above application, the drug for preventing or treating ulcerative colitis also contains a pharmaceutically acceptable carrier and / or excipient.

[0026] Furthermore, in the above application, the auxiliary material is one or more of an antioxidant, a preservative, a solubilizer, a disintegrant, a lubricant, a colorant, a dispersant, and a surfactant.

[0027] Furthermore, in the above application, the administration method of the drug for preventing or treating ulcerative colitis is preferably gastrointestinal administration.

[0028] Furthermore, in the treatment of ulcerative colitis, the drug is used to reduce the levels of C3c, TNF-α, and IL-6 in serum.

[0029] Further, in the treatment of ulcerative colitis, the medicament is used to increase IL-10 levels.

[0030] Furthermore, in the treatment of ulcerative colitis, the drug is used to improve colon tissue damage.

[0031] Further, in the treatment of ulcerative colitis, the medicament is used to reduce the degree of colon shortening and spleen index.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] The present invention applies the ethanol extract and flavonoids of wild horse herb to drugs related to the prevention and treatment of ulcerative colitis for the first time, and verifies it by constructing a mouse ulcerative colitis model. The results show that the flavonoids of wild horse herb and the ethanol extract of wild horse herb can improve the level of inflammation in mice with ulcerative colitis and inhibit the complement level; and can improve colon tissue damage, significantly reduce the DAI score, reduce the degree of colon shortening, and alleviate the symptoms of colon congestion and swelling; in addition, it can also relieve spleen enlargement and reduce spleen index. The present invention not only verifies that the flavonoids of wild horse herb and the ethanol extract of wild horse herb have the effect of resisting ulcerative colitis through animal experiments, but also studies the mechanism of action of the flavonoids of wild horse herb and the ethanol extract of wild horse herb on ulcerative colitis, providing a scientific basis for the use of the flavonoids of wild horse herb and the ethanol extract of wild horse herb for the prevention and treatment of ulcerative colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 This is the HPLC spectrum of the flavonoids of Yemazhui, where ① is the chromatographic peak corresponding to Zedoaria lutein (7.940), and ② is the chromatographic peak corresponding to cyanidin (13.880);

[0035] Figure 2 The curves of DAI scores of mice in different experimental groups are shown. Compared with the blank group, ####P<0.0001; compared with the DSS model group, *P<0.05, ***P<0.001, ****P<0.0001, n=6;

[0036] Figure 3 is the colon length of mice in different experimental groups, among which, compared with the blank group, #### P<0.0001; compared with the DSS model group, *P<0.05, ***P<0.001, ****P<0.0001, n=6;

[0037] Figure 4 is the spleen index of mice in different experimental groups, among which, compared with the blank group, #### P<0.0001; compared with the DSS model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, n=6;

[0038] Figure 5 is the C3c content in the serum of mice in different experimental groups. Compared with the blank group, #### P<0.0001; compared with the DSS model group, *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, n=6;

[0039] Figure 6 is the content of IL-6, IL-10 and TNF-α in the serum of mice in different experimental groups. Compared with the blank group, #### P<0.0001; compared with the DSS model group, **P<0.01, ****P<0.0001, n=6;

[0040] Figure 7 These are the H&E staining images of the colon tissues of mice in different experimental groups, a: blank group, b: model group, c: Yemazhui flavonoid part group, d: positive drug group. DETAILED DESCRIPTION

[0041] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art of the present invention. The terms used herein in the specification of the present invention are only for the purpose of describing specific embodiments and are not intended to limit the present invention. The term "and / or" used herein includes any and all combinations of one or more related listed items.

[0042] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments so that those skilled in the art can better understand the present invention and implement it, but the embodiments are not intended to limit the present invention.

[0043] Example

[0044] 1. Purpose of the experiment

[0045] To observe the effects of the flavonoids of Rhizoma Cibotii and its ethanol extract on the pharmacological indices of mice with ulcerative colitis, including DAI score, colon length, spleen index, colon tissue damage, serum inflammatory factor levels and complement.

[0046] 2. Experimental Materials

[0047] Experimental medicinal materials and reagents: Yemazhui medicinal materials were purchased from Xuyi Defeng Chinese Medicinal Materials Planting Company in Huai'an City; dextran sulfate sodium was purchased from Shanghai Xibao Biotechnology Co., Ltd.; toluene, ammonia water, hydrochloric acid, anhydrous ethanol and neutral gum were purchased from Sinopharm Chemical Reagent Co., Ltd.; sulfasalazine enteric-coated tablets were purchased from Shanghai Xinyi Tianping Pharmaceutical Co., Ltd.; ELISA kits for TNF-α, IL-10, IL-6 and C3c were purchased from Shanghai Meike Biotechnology Co., Ltd.; other chemical reagents were of analytical grade.

[0048] Experimental instruments: 702-type -80℃ refrigerator, produced by Thermo Fisher Scientific Inc., USA; TECAN full-wavelength microplate reader, model M1000 pro, produced by Nanjing Kelinde Scientific Instrument Co., Ltd.; incubator, produced by Shanghai Jinghong Experimental Equipment Co., Ltd.; balance, produced by Ohaus Instrument Co., Ltd.; water bath, produced by Shanghai Ailan Instrument Co., Ltd.; high performance liquid chromatograph, model Agilent 1260Infinity, produced by Agilent Technologies, USA.

[0049] 3. Experimental Methods

[0050] Preparation of the ethanol extract of Z. truncatum: weigh 100 g of dried whole herb of Z. truncatum, cut it into 2-3 cm segments, add 10 times the volume of 80% ethanol aqueous solution, reflux extraction for 2 hours, repeat 2 times, recover the solvent under reduced pressure at 40°C, and obtain 15 g of Z. truncatum ethanol extract (ZTW).

[0051] Preparation of the flavonoid fraction of Herba Aspergillus: The ethanol extract of Herba Aspergillus was dissolved in about 90 mL of water, adsorbed by 170 mL of D101 macroporous adsorption resin, firstly eluted with 1700 mL of water to remove macromolecular compounds, then eluted with 1700 mL of 95% ethanol aqueous solution by volume, and the solvent was recovered under reduced pressure to obtain 5 g of the macroporous resin fraction; the obtained macroporous resin fraction was adsorbed by 120 mL of YMCODS-A reverse-phase silica gel filler with a particle size of 60 μm reverse-phase silica gel chromatography column, eluted with 550 mL of 50% methanol aqueous solution by volume, and then eluted with 550 mL of 70% methanol aqueous solution, and the solvent was recovered under reduced pressure at 40° C. to obtain 2 g of the extract, and the extract was then adsorbed by 50 mL of 30-60 mesh polyamide chromatography column, firstly eluted with 200 mL of 70% ethanol aqueous solution by volume, and then eluted with 200 mL of 95% ethanol, and the solvent was recovered under reduced pressure at 40° C. to obtain 0.2 g of the flavonoid fraction of Herba Aspergillus (SHT).

[0052] The components of the flavonoids of YEMAZHUAI were determined by HPLC, and the HPLC conditions were as follows: mobile phase A was acetonitrile (containing 0.1% formic acid); mobile phase B was water (containing 0.1% formic acid); mobile phase conditions: 0 min: 36% A, 15 min: 36% A, 30 min: 90% A; chromatographic column: YMC-PACK ODS-A (Ф4.6 mm×250 mm, 5 μm); column temperature: 28°C; detection wavelength: 365 nm; injection volume: 20 μL; flow rate: 1 mL·min -1 .

[0053] Weigh 4.67 mg of reference substance Zeolite lutein and 8.17 mg of brown cyanidin respectively; and use mobile phase A to make it up to 10 mL as the stock solution, add mobile phase A to dilute it to 1:2, 1:4, 1:8, 1:16 and 1:32, filter it with a 0.22 μm filter membrane, and set aside the filtrate. Take 5 concentrations of reference substances for chromatographic determination, and draw a standard curve. Each concentration point is analyzed in parallel 3 times. Record the peak area of ​​each chromatographic peak, and linearly regress its concentration (x) with the peak area (y) of the reference substance to obtain the standard curve of the reference substance, Zeolite lutein y = 52964x + 19.30, brown cyanidin y = 55161x + 210.79. 6 mg of the sample of the flavonoid part (SHT) of Yemazhui was made up to 10 ml volumetric flask with mobile phase A, filtered with a 0.22 μm filter membrane, and the filtrate was set aside. The filtrate was taken for chromatographic determination.

[0054] like Figure 1 As shown, after high performance liquid chromatography analysis, it was found that the sample of the flavonoid part of Houttuynia cordata contained zedoary lutein and brown cyanidin. The chromatographic signal values ​​of the samples were substituted into the standard curve to further obtain the specific content of each component. The flavonoid part of Houttuynia cordata (SHT) contained 25.2% zedoary lutein and 65.7% brown cyanidin, and the total proportion was 90.9%.

[0055] Animal grouping, modeling and drug administration: Male C57BL / 6J mice were divided into blank group (Control), model group (Model), ZTW ethanol extract group, ZTW flavonoids group (SHT), and positive drug group (Positive, sulfasalazine) with 6 mice in each group after adaptive culture. Except for the blank group, the model group, ZTW ethanol extract group, ZTW flavonoids group and positive drug group were all given 2.5% DSS solution for drinking, and the 2.5% DSS solution was changed every 3 days for 7 consecutive days. From the second day, ZTW total extract, ZTW flavonoids group and positive drug were given oral gavage treatment, the blank group was given normal saline for 8 consecutive days, the ZTW ethanol extract group and ZTW flavonoids group were both given a dose of 0.2g / kg, and the positive drug group was given a dose of 0.25g / kg.

[0056] Determination of pharmacological indicators: According to the Cooper scoring system (as shown in Table 1 below), the mouse colon tissue disease activity index (DAI) was assessed daily based on the weight loss, stool characteristics, and blood in the stool of the mice. On the 9th day, half an hour after the last administration, blood was collected from the mouse eyeball, stored at 4°C, centrifuged at 3500rpm for 6 minutes, and serum was collected and stored at -80°C for IL-6, IL-10, TNF-α and C3c determination; the mouse colon was collected for measuring the colon length; the mouse colon tissue was collected in paraformaldehyde fixative for H&E staining observation; the spleen was collected for weighing and the spleen index was calculated.

[0057] Table 1 Disease Activity Index (DAI) Scoring Criteria

[0058]

[0059] 4. Experimental Results

[0060] The following experimental data results are expressed as mean ± standard deviation (x ± SD). SPSS 21.0 was used to perform one-way analysis of variance on multiple groups of measurement data, and t-test was used for inter-group comparison. P < 0.05 was considered statistically significant. SPSS 21.0 was used to perform Turkey-HSD rank sum test on OUT.

[0061] DAI score: Figure 2 As shown, on the last day of modeling, the DAI score of mice in the model group was significantly increased compared with that in the blank group (P<0.0001); 8 days after oral administration, compared with the model group, the DAI score of mice in the ethanol extract group of R. truncatum (ZTW in the figure) and the flavonoid part group of R. truncatum (SHT in the figure) was significantly reduced (P<0.0001), and was better than that in the positive drug group (Positive in the figure).

[0062] Colon length: Figure 3 As shown, compared with the blank group, the colon length of mice in the model group was significantly shortened (P<0.0001); compared with the model group, the degree of colon shortening of mice in the ethanol extract of Echinops edulis group (ZTW in the figure) and the flavonoid part of Echinops edulis group (SHT in the figure, P<0.05) was reduced, but the data of the ethanol extract of Echinops edulis group were not statistically significant.

[0063] Spleen index: Figure 4 As shown, compared with the blank group, the spleen index of mice in the DSS model group was significantly increased (P<0.0001); compared with the DSS model group, the spleen index of mice in the Mustang ethanol extract group (ZTW in the figure, P<0.05), Mustang flavonoids group (SHT in the figure) and positive drug group (Positive in the figure) were all decreased, but the data of the Mustang flavonoids group (SHT in the figure) and the positive drug group (Positive in the figure) were not statistically significant.

[0064] Serum complement C3c level: Figure 5 As shown, compared with the blank group, the complement C3c level in the serum of mice in the DSS model group was significantly increased (P<0.0001); compared with the DSS model group, the complement C3c level in the serum of mice in the ethanol extract of Y. machus group (ZTW in the figure, P<0.01) and the flavonoid part of Y. machus group (SHT in the figure, P<0.001) was significantly decreased.

[0065] Serum inflammatory factor levels: Figure 6 As shown in the figure, the levels of serum TNF-α and IL-6 in the model group mice were significantly higher than those in the blank group (P<0.0001), while the level of IL-10 was significantly lower than that in the blank group (P<0.0001). Compared with the model group, the levels of serum TNF-α and IL-6 in the wild horse herb ethanol extract group (ZTW in the figure, IL-6: P<0.0001, TNF-α: P<0.0001) and the wild horse herb flavonoid part group (SHT in the figure, IL-6: P<0.0001, TNF-α: P<0.0001) were significantly reduced, and the level of IL-10 was significantly increased (P<0.0001).

[0066] H&E staining of colon tissue: H&E staining of colon tissue of mice in different experimental groups is shown in the figure below. Figure 7 As shown in the figure, the intestinal tissue structure of the mice in the model group was severely abnormal, with some epithelial cells in the mucosal layer eroded and shed, and the lamina propria exposed. Figure 7 b Arrow 1; some crypts can be seen to be slightly expanded and epithelial cells are atrophied, such as Figure 7 b Arrow 2 shows that the number of goblet cells is significantly reduced. Figure 7 b As shown by arrow 3, focal infiltration of inflammatory cells can be seen in the tissue. Figure 7 b As shown by arrow 4. The intestinal tissue structure of the mice in the group of flavonoids from Yemazhui was slightly abnormal. The mucosal layer structure was intact and neatly arranged, and no necrosis or shedding of epithelial cells was observed. The crypt structure was normal, tightly arranged, and no expansion was observed. The number of goblet cells was abundant and neatly arranged. A small amount of inflammatory cell infiltration was observed in the tissue, such as Figure 7 c As shown by arrow 1. The intestinal tissue structure of mice in the positive drug group was slightly abnormal, with some epithelial cells in the mucosal layer eroded and fell off, and the lamina propria exposed, as shown in Figure 2. Figure 7 d As shown by arrow 1; the crypt structure is normal, arranged tightly, and no expansion is observed; the goblet cells are abundant and arranged regularly and tightly; no obvious inflammatory cell infiltration is observed in the tissue.

[0067] 5. Conclusion

[0068] The above experimental results show that the flavonoid part of wild horse herb and the ethanol extract group of wild horse herb can reduce the levels of C3c, TNF-α, and IL-6 in the serum of ulcerative colitis mice, increase the level of IL-10, improve the inflammation level in the body of ulcerative colitis mice, inhibit the complement level, improve the colon tissue damage, significantly reduce the DAI score, reduce the degree of colon shortening, and alleviate the symptoms of colon congestion and swelling; relieve splenomegaly and reduce the spleen index.

[0069] The above-described embodiments are only preferred embodiments for fully illustrating the present invention, and the protection scope of the present invention is not limited thereto. Equivalent substitutions or changes made by those skilled in the art based on the present invention are within the protection scope of the present invention. The protection scope of the present invention shall be subject to the claims.

Claims

1. The use of the flavonoids of Yemazhui in the preparation of a drug for preventing or treating ulcerative colitis, characterized in that: The preparation of the flavonoid fraction of Yemazhui comprises the following steps: (1) Take the dried whole herb of Psoralea corylifolia, cut it into 2-3 cm segments, add 8-15 times the volume of 60%-95% ethanol aqueous solution, reflux extraction for 1-3 hours, and recover the solvent under reduced pressure at 40-60 °C to obtain Psoralea corylifolia ethanol extract; (2) dissolving the ethanol extract of the wild horsetail prepared in step (1) in water to prepare a 0.1-0.2 g / mL solution, adsorbing it on a weakly polar macroporous adsorption resin, first eluting it with water to remove macromolecular compounds, then eluting it with a 90%-95% ethanol aqueous solution by volume, and recovering the solvent under reduced pressure at 40-60° C. to obtain a macroporous resin fraction; (3) The macroporous resin portion prepared in step (2) is adsorbed on an ODS reverse phase silica gel column, eluted with a 50% by volume methanol aqueous solution, then eluted with a 70%-75% by volume methanol aqueous solution, and the solvent is recovered under reduced pressure at 40-60°C to obtain an extract. The extract is then adsorbed on a 30-60 mesh polyamide column, first eluted with a 70% by volume ethanol aqueous solution, then eluted with a 95% by volume ethanol aqueous solution, and the solvent is recovered under reduced pressure at 40-60°C to obtain the flavonoid portion of the wild horsetail.

2. The use according to claim 1, characterized in that: The flavonoid fraction of YEMATU refers to the flavonoid fraction of YEMATU containing cyanidin and lutein.

3. The use according to claim 1, characterized in that: In step (2), the weakly polar macroporous adsorption resin is AB-8 macroporous adsorption resin.

4. The use according to claim 1, characterized in that: In step (3), the ODS reverse-phase silica gel chromatographic column is a DAISOGEL ODS reverse-phase silica gel chromatographic column, a Cosmsil ODS reverse-phase silica gel chromatographic column or a YMCODS-A reverse-phase silica gel chromatographic column; the filler particle size of the ODS reverse-phase silica gel chromatographic column is 50-70 μm.

5. The use according to claim 1, characterized in that: In step (3), the flavonoid fraction of the wild horsetail contains 63%-68% of cyanidins and 24%-29% of luteolin.

6. The use according to claim 1, characterized in that: The medicine further comprises a pharmaceutically acceptable carrier and / or excipient; the excipient is one or more of an antioxidant, a preservative, a solubilizer, a disintegrant, a lubricant, a colorant, a dispersant, and a surfactant.

7. The use according to claim 1, characterized in that: The administration method of the drug for preventing or treating ulcerative colitis is gastrointestinal administration.

8. Use of the ethanol extract of Rhizoma Cibotii in the preparation of a drug for preventing or treating ulcerative colitis, characterized in that: The ethanol extract of Mustard Seedlings is an extract obtained by treating Mustard Seedlings with a 60%-95% ethanol aqueous solution.

9. The use according to claim 8, characterized in that: The preparation method of the ethanol extract of Echinops chinensis is as follows: taking the dried whole herb of Echinops chinensis, cutting it into 2-3 cm segments, adding 8-15 times the volume of 60%-95% ethanol aqueous solution, refluxing for 1-3 hours, and recovering the solvent under reduced pressure at 40-60° C. to obtain the ethanol extract of Echinops chinensis.

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