A traditional Chinese medicine composition for treating ulcerative colitis, a preparation method and use thereof

Abstract

This invention relates to the field of traditional Chinese medicine (TCM) technology, and discloses a TCM composition for treating ulcerative colitis, its preparation method, and its uses. The TCM composition comprises the following raw materials in parts by weight: 2-10 parts of charred Typhae Pollen, 3-7 parts of charred Scutellariae Radix, 4-6 parts of charred Angelicae Sinensis Radix, 3-8 parts of charred Sophorae Fructus, 6-10 parts of Prunus Mume, 2-10 parts of prepared Zingiberis Rhizoma, 5-7 parts of prepared Aconitum Carmichaelii Radix, 3-7 parts of Coptise Rhizoma, and 1-8 parts of Indigo Naturalis Radix. Among these, Prunus Mume is the principal ingredient, with prepared Zingiberis Rhizoma and prepared Aconitum Carmichaelii Radix as assistant ingredients, and charred Angelicae Sinensis Radix, charred Sophorae Fructus, charred Typhae Pollen, charred Scutellariae Radix, Coptise Rhizoma, and Indigo Naturalis as adjuvants. The synergistic effect among the various TCM components gives the TCM composition of this invention the effects of soothing the liver and strengthening the spleen, astringing the intestines and stopping bleeding. It is mainly used to treat moderate to severe ulcerative colitis. Experiments have demonstrated that the traditional Chinese medicine composition of this invention can inhibit intestinal inflammatory response in mice with ulcerative colitis; improve pathological histological damage and promote intestinal mucosal repair; regulate macrophage polarization and restore the balance of M1 / M2 macrophages; regulate CD4+ T cell differentiation and reshape Th17 / Treg cell immune homeostasis.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine technology, specifically to a traditional Chinese medicine composition for treating ulcerative colitis, its preparation method, and its uses. Background Technology

[0002] Ulcerative colitis (UC) is a chronic, nonspecific inflammatory disease of unknown cause that primarily affects the colorectal mucosa and submucosa. Currently, immune dysregulation is considered the main mechanism of UC pathogenesis. Therefore, 5-aminosalicylic acid preparations (5-ASA) are currently first-line drugs for the treatment of mild to moderate ulcerative colitis. They can affect the synthesis of inflammatory factors (prostaglandins, leukotrienes, etc.) in the intestinal mucosa and submucosa, thereby regulating the intestinal immune response, inhibiting inflammatory reactions, and protecting the intestinal mucosa.

[0003] Patients with moderate to severe ulcerative colitis and more pronounced clinical symptoms should consider step-up therapy using steroids, immunosuppressants, or biologics. Steroids are commonly used clinically to treat moderate to severe ulcerative colitis or 5-ASA failure, with a clinical remission rate of approximately 36%. Long-term use can cause adverse reactions such as moon face, acne, insomnia, and decreased immunity. Immunosuppressants are suitable for ulcerative colitis patients who are unresponsive to or dependent on steroids, or intolerant to aminosalicylic acid preparations, and as a "rescue" treatment for severe ulcerative colitis patients who do not respond to adequate intravenous steroid therapy. However, long-term use can cause serious consequences such as liver and kidney toxicity and bone marrow suppression. Biologics also target the immune response, with a clinical remission rate of approximately 15-40% in ulcerative colitis patients. However, studies have found that the use of biologics may significantly increase the risk of complications such as tuberculosis, tumors, and opportunistic infections. Therefore, while current medications for ulcerative colitis can modulate the immune response and achieve some therapeutic effect, they suffer from limitations in the types of drugs available, unstable efficacy, high relapse rates, and significant side effects. Thus, for patients with moderate to severe ulcerative colitis, there is an urgent need to find a treatment method that can effectively induce remission, is highly safe, and has fewer adverse reactions. Summary of the Invention

[0004] In view of this, the present invention provides a traditional Chinese medicine composition for treating ulcerative colitis, its preparation method and uses, to solve the problems of limited drug types, unstable efficacy, easy relapse and obvious toxic side effects in the step-up treatment of moderate to severe patients using steroid hormones, immunosuppressants or biological agents.

[0005] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating ulcerative colitis.

[0006] In one optional embodiment, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 2-10 parts of charred cattail pollen, 3-7 parts of charred scutellaria baicalensis, 4-6 parts of charred angelica sinensis, 3-8 parts of charred sophora japonica, 6-10 parts of dried plum, 2-10 parts of prepared ginger, 5-7 parts of black aconite, 3-7 parts of coptis chinensis, and 1-8 parts of indigo naturalis.

[0007] In one optional embodiment, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 4-6 parts of charred cattail pollen, 5-7 parts of charred scutellaria baicalensis, 4-6 parts of charred angelica sinensis, 5-7 parts of charred sophora japonica, 6-10 parts of dried plum, 4-8 parts of prepared ginger, 5-7 parts of black aconite, 3-7 parts of coptis chinensis, and 2-5 parts of indigo naturalis.

[0008] In one optional embodiment, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 5 parts of charred cattail pollen, 6 parts of charred scutellaria baicalensis, 5 parts of charred angelica sinensis, 6 parts of charred sophora japonica, 8 parts of dried plum, 6 parts of prepared ginger, 6 parts of black aconite root slices, 5 parts of coptis chinensis, and 3 parts of indigo naturalis.

[0009] Secondly, the present invention provides a method for preparing a traditional Chinese medicine composition for treating ulcerative colitis. In an optional embodiment, the method for preparing the traditional Chinese medicine composition for treating ulcerative colitis includes: weighing raw materials according to a specified ratio, mixing all raw materials and then extracting them using conventional extraction methods, or extracting them separately using conventional extraction methods and then mixing them to obtain the final product.

[0010] In one optional embodiment, the conventional extraction method includes one or more of the following: decoction extraction, maceration extraction, percolation extraction, reflux extraction, ultrasonic extraction, and steam distillation extraction; the extraction is performed at least once; and each extraction takes at least 10 minutes.

[0011] And / or, the extraction solvent is selected from water or an alcohol solution with a volume percentage of 5-98%;

[0012] And / or, the weight of the extraction solvent is more than three times the total weight of the raw materials;

[0013] And / or, the temperature of the water extraction is 90-100℃.

[0014] In one optional implementation, the conventional extraction method specifically includes:

[0015] (1) Weigh the raw materials according to the proportion, mix all the raw materials and add water 3-10 times the total weight of the raw materials to form a medicinal solution;

[0016] (2) Heat the medicinal liquid to 90-100℃, then keep it at 90-100℃ for 0.5-1 hour, filter, and concentrate the filtrate to obtain the product;

[0017] Optionally, in step (2), the medicinal solution is soaked for 1-2 hours before heating;

[0018] The concentration includes: concentrating the filtrate to 1 / 3-2 / 3 of its original volume at 50-70°C;

[0019] Alternatively, the conventional extraction method specifically includes:

[0020] (1) Weigh the raw materials according to the ratio, add the prepared ginger, black aconite slices, coptis and indigo to 3-10 times the weight of the raw materials in an ethanol solution with a volume percentage of 5-98%, extract at room temperature for 30-50 minutes by ultrasonic extraction, and then mix the extracts to form mixed drug solution A.

[0021] (1) Mix the charred cattail pollen, charred scutellaria baicalensis, charred angelica sinensis, charred sophora japonica and dried plum, and add water at 3-10 times the total weight of the mixed raw materials to form mixed medicine solution B;

[0022] (2) Mix the mixed medicine solution B and the mixed medicine solution A and heat to 90-100℃, then keep warm at 90-100℃ for 0.5-1 hour, filter, and concentrate the filtrate to obtain the product;

[0023] Optionally, the concentration includes concentrating the filtrate to 1 / 3-2 / 3 of its original volume at 50-70°C.

[0024] Thirdly, the present invention provides a traditional Chinese medicine preparation.

[0025] In one optional embodiment, the active ingredient of the traditional Chinese medicine preparation is the traditional Chinese medicine composition prepared by the method described above or by the method described above.

[0026] Fourthly, the present invention provides the use of the traditional Chinese medicine composition, the traditional Chinese medicine composition prepared by the preparation method of the traditional Chinese medicine composition, or the traditional Chinese medicine preparation described herein in the preparation of a medicament for relieving, adjuvant treatment, or treatment of ulcerative colitis.

[0027] In one alternative embodiment, the drug for relieving, adjunctive treatment, or treating ulcerative colitis includes at least one of the following effects (1)-(6):

[0028] (1) Soothes the liver and strengthens the spleen, and / or astringes the intestines and stops bleeding;

[0029] (2) Inhibit intestinal inflammatory response and reduce the degree of inflammatory cell infiltration;

[0030] (3) Improve pathological and histological damage and promote the repair of intestinal mucosal barrier function;

[0031] (4) Regulate macrophage polarization and restore the M1 / M2 macrophage balance;

[0032] (5) Regulate CD4+ T cell differentiation and reshape Th17 / Treg cell immune homeostasis.

[0033] In one alternative embodiment, the ulcerative colitis is ulcerative colitis caused by liver yin deficiency;

[0034] And / or, the ulcerative colitis is ulcerative colitis caused by liver stagnation or liver reversal;

[0035] Optionally, the ulcerative colitis is moderate to severe ulcerative colitis.

[0036] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0037] This invention provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following raw materials in parts by weight: 2-10 parts of charred cattail pollen, 3-7 parts of charred scutellaria baicalensis, 4-6 parts of charred angelica sinensis, 3-8 parts of charred sophora japonica, 6-10 parts of dried plum, 2-10 parts of prepared ginger, 5-7 parts of prepared aconite root, 3-7 parts of coptis chinensis, and 1-8 parts of indigo naturalis. In this traditional Chinese medicine composition, dried plum is the principal ingredient. On one hand, its sour and sweet properties nourish the body fluids, thus nourishing yin and softening the liver. With the liver nourished, yin and blood gradually recover, and the spleen and stomach strengthen. On the other hand, its sour and astringent properties consolidate the intestines, thus stopping diarrhea. Thirdly, its sour and sweet properties nourish yin, relieve spasms and pain, thus alleviating abdominal pain. Prepared ginger and prepared aconite root are used as assistant ingredients; both are warming and tonifying herbs, which can assist the principal ingredient, dried plum, in strengthening its warming and spleen-strengthening effects. With the spleen and stomach strengthened, dampness is eliminated, and diarrhea stops. At the same time, prepared ginger can also warm the meridians and relieve pain, assisting dried plum in enhancing its ability to stop abdominal pain. Prepared ginger itself also has the effect of strengthening the spleen and stopping bleeding, and can be used to stop bleeding. The formula is supplemented with charred angelica root, charred sophora japonica flower, charred cattail pollen, charred scutellaria baicalensis root, coptis chinensis root, and indigo naturalis. Angelica root is sweet and pungent in taste, warm in nature, and enters the liver, heart, and spleen meridians. It can enhance the blood-nourishing and liver-soothing effects of dried plum, while the charred form enhances the hemostatic effect of roasted ginger. Charred sophora japonica flower, charred cattail pollen, and charred scutellaria baicalensis root can help dried plum soothe the liver and clear heat, as well as clear the intestines and stop bleeding. Coptis chinensis root and indigo naturalis root are used to cool the blood and stop bleeding. The synergistic effect of the various Chinese herbal components gives the herbal composition of this invention the effects of soothing the liver and strengthening the spleen, astringing the intestines and stopping bleeding. It can be used to treat ulcerative colitis, especially for moderate to severe ulcerative colitis.

[0038] Experiments have demonstrated that the traditional Chinese medicine composition of this invention can inhibit intestinal inflammatory response in mice with ulcerative colitis, reduce the degree of inflammatory cell infiltration, improve pathological histological damage, promote intestinal mucosal repair, regulate macrophage polarization, restore the balance of M1 / M2 macrophages, regulate CD4+ T cell differentiation, and reshape Th17 / Treg cell immune homeostasis. Attached Figure Description

[0039] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0040] Figure 1 This describes the changes in body weight of mice in each group in the experimental examples of this invention;

[0041] Figure 2 These are the DAI scores of mice in each group in the experimental examples of this invention;

[0042] Figure 3 This describes the colon length of mice in each group in the experimental examples of this invention;

[0043] Figure 4 This refers to the ratio of spleen weight to body weight in each group of mice in the experimental examples of this invention;

[0044] Figure 5 These are the HE staining and pathological scores of mice in each group in the experimental examples of this invention; where A: HE staining of mice in each group (×200); B: pathological histological scores of mice in each group.

[0045] Figure 6 This invention compares the regulatory effects of M1 macrophages in different groups of UC mice in the experimental examples; where A: M1 macrophages in UC mice; B: Analysis of the content level of M1 macrophages in UC mice.

[0046] Figure 7 This is a comparison of the regulatory effects of M2 macrophages in different groups of UC mice in the experimental examples of this invention; where A: M2 macrophages in UC mice; B: analysis of the content level of M2 macrophages in UC mice.

[0047] Figure 8 This describes the regulation of Th17 cells in each group of UC mice in the experimental examples of this invention; where A: Th17 cells in UC mice; B: Th17 cell content analysis in UC mice;

[0048] Figure 9 This describes the regulation of Treg cells in UC mice in each group of experimental examples of this invention; A: Treg cells in UC mice; B: Analysis of Treg cell content levels in UC mice. Detailed Implementation

[0049] The following embodiments are provided to better understand the present invention and are not limited to the preferred embodiments described. They do not constitute a limitation on the content and scope of protection of the present invention. Any product that is the same as or similar to the present invention, derived by any person under the guidance of the present invention or by combining the features of the present invention with other prior art, falls within the protection scope of the present invention.

[0050] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.

[0051] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the application.

[0052] Currently, 5-aminosalicylic acid (5-ASA) is the first-line drug for treating mild to moderate ulcerative colitis, while patients with moderate to severe UC need to use step-up therapy such as corticosteroids, immunosuppressants or biologics. However, these treatments have problems such as limited drug types, unstable efficacy, easy relapse, and significant toxic side effects.

[0053] Active ulcerative colitis can be classified into mild, moderate, and severe according to clinical severity. The modified Mayo Criterion (MCC) is commonly used to assess clinical severity and treatment efficacy. The MCC criteria are based on the "Consensus Opinion on the Integrated Traditional Chinese and Western Medicine Diagnosis and Treatment of Ulcerative Colitis (2017)," the "Guidelines for the Diagnosis and Treatment of Ulcerative Colitis with Traditional Chinese Medicine (2023)," and the "Guidelines for the Diagnosis and Treatment of Ulcerative Colitis in China (2023·Xi'an)," as detailed in the table below:

[0054]

[0055] Note: a) Each subject serves as a self-control to evaluate the degree of abnormality in bowel movement frequency; b) Daily bleeding score, representing the most severe bleeding in a day; c) Physician's overall evaluation includes three criteria: subject's review of abdominal discomfort, overall well-being, and other manifestations such as physical examination findings and subject performance status.

[0056] Based on years of clinical experience, the inventors of this application have discovered that mild to moderate ulcerative colitis is often due to spleen deficiency, leading to impaired digestion and absorption, internal fluid production, and the downward movement of fluids into the intestines, resulting in damp-heat and damage to the intestinal network. For moderate to severe ulcerative colitis, it is necessary to further focus on the root cause of spleen deficiency. According to the Five Elements theory in Traditional Chinese Medicine, the spleen belongs to Earth, and the liver belongs to Wood; the two have a mutually restraining relationship. If the Liver Wood element is excessive and suppresses the Spleen Earth element, it will lead to spleen and stomach weakness. In our clinical practice, we have found that patients with moderate to severe ulcerative colitis often suffer from excessive worry, excessive sleep deprivation, or qi stagnation leading to internal heat, which damages the liver and depletes liver yin. Insufficient liver yin results in a lack of nourishment, leading to liver stagnation or liver qi reversal due to insufficient material support for the liver's function of regulating qi. This weakens the spleen and stomach in the middle jiao, causing internal phlegm and dampness, which, over time, transforms into heat. This damp-heat descends to the intestines, damaging the intestinal vessels and resulting in diarrhea and bloody, mucous stools, ultimately developing into ulcerative colitis. Furthermore, insufficient liver yin deprives the body's vessels of nourishment, leading to pain due to lack of nourishment, which can manifest as abdominal pain, or abdominal pain and diarrhea with pain relieved after defecation—common symptoms of ulcerative colitis. Therefore, the treatment of this disease should focus on the liver, employing methods such as soothing the liver, strengthening the spleen, and astringing the intestines to stop bleeding.

[0057] Based on this, according to a first aspect of the present invention, the present invention provides a traditional Chinese medicine composition for treating ulcerative colitis.

[0058] In some embodiments, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 2-10 parts of charred cattail pollen, 3-7 parts of charred scutellaria baicalensis, 4-6 parts of charred angelica sinensis, 3-8 parts of charred sophora japonica, 6-10 parts of dried plum, 2-10 parts of prepared ginger, 5-7 parts of black aconite, 3-7 parts of coptis chinensis, and 1-8 parts of indigo naturalis.

[0059] In the traditional Chinese medicine composition of the present invention, smoked plum is the monarch drug. Smoked plum is sour, astringent, and neutral in nature, and belongs to the liver, spleen, lung, and large intestine meridians. "Compendium of Materia Medica" records that it can "astringe the lung and arrest diarrhea, stop chronic cough and dysentery". "Explanation of the Divine Farmer's Classic of Materia Medica" records that "smoked plum is sour... sour can astringe deficient fire, transform body fluids, and secure the prolapsed intestine". Therefore, smoked plum is selected in the present invention, which has three functions: on the one hand, sour and sweet can generate body fluids, nourish the liver and soften the liver, so that the liver can be nourished, yin and blood gradually recover, and the spleen and stomach become stronger; on the other hand, sour astringency can secure the prolapsed intestine and arrest diarrhea; thirdly, sour and sweet can nourish yin and relieve spasm and pain, and relieve abdominal pain. The ministerial drugs are prepared ginger and processed aconite root. Prepared ginger belongs to the spleen, stomach, and kidney meridians, and can warm the middle-jiao to stop pain and strengthen the spleen to stop bleeding; processed aconite root belongs to the heart, kidney, and spleen meridians, and can reinforce yang to promote fire and warm the middle-jiao to stop diarrhea. The two drugs can assist the monarch drug smoked plum to strengthen the function of warming yang and strengthening the spleen. Once the spleen and stomach are strengthened, dampness and water will naturally disappear, and diarrhea will stop; at the same time, prepared ginger can also warm the meridians to stop pain and assist smoked plum to enhance the power of relieving abdominal pain; prepared ginger itself also has the effect of strengthening the spleen to control bleeding and can be used to stop bleeding. The adjuvant drugs are carbonized angelica, carbonized sophora flower, carbonized cattail pollen, carbonized scutellaria baicalensis, coptis chinensis, and indigo naturalis. Angelica is sweet, pungent, and warm in nature, and belongs to the liver, heart, and spleen meridians. It can not only assist smoked plum to strengthen the function of nourishing blood and softening the liver, but also char to form carbon, enhancing the hemostatic effect of prepared ginger. Carbonized sophora flower belongs to the liver and large intestine meridians, cooling blood to stop bleeding and clearing the liver to purge fire; carbonized cattail pollen belongs to the liver meridian and pericardium meridian; cooling blood to stop bleeding and activating blood circulation to remove stasis; carbonized scutellaria baicalensis belongs to the lung meridian, gallbladder meridian, spleen meridian, large intestine meridian, and small intestine meridian, and can purge excessive fire, remove damp-heat, and stop bleeding. The combination of the three drugs can not only assist smoked plum to soften the liver and purge fire, but also clear the intestine and stop bleeding; coptis chinensis belongs to the heart, spleen, stomach, liver, gallbladder, and large intestine meridians, and can clear heat and dry dampness, purge fire and detoxify; indigo naturalis belongs to the liver meridian, and can clear heat and detoxify, cool blood to astringe sores, purge fire and calm fright. The two are combined to cool blood to stop bleeding. In summary, the above-mentioned medicinal materials are combined, and the various traditional Chinese medicine components synergistically enhance the effect, making the traditional Chinese medicine composition of the present invention have the effects of softening the liver and strengthening the spleen, astringing the intestine and stopping bleeding. Combining with the pathogenesis characteristics of liver yin deficiency, damp-heat stasis and binding in moderate and severe ulcerative colitis, this formula has good curative effects on moderate and severe ulcerative colitis (moderate and severe ulcerative colitis).

[0060] Moreover, it has been experimentally proven that through the effects of softening the liver and strengthening the spleen, astringing the intestine and stopping bleeding, the traditional Chinese medicine composition of the present invention can not only inhibit the inflammatory response of ulcerative colitis mice, promote mucosal repair, but also regulate the polarization balance of M1 / M2 macrophages and reshape the immune homeostasis of Th17 / Treg cells, so as to achieve the purpose of treating ulcerative colitis.

[0061] In some embodiments, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 4-6 parts of carbonized cattail pollen, 5-7 parts of carbonized scutellaria baicalensis, 4-6 parts of carbonized angelica, 5-7 parts of carbonized sophora flower, 6-10 parts of smoked plum, 4-8 parts of prepared ginger, 5-7 parts of processed aconite root, 3-7 parts of coptis chinensis, and 2-5 parts of indigo naturalis.

[0062] In some embodiments, the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 5 parts of charred cattail pollen, 6 parts of charred scutellaria baicalensis, 5 parts of charred angelica sinensis, 6 parts of charred sophora japonica, 8 parts of dried plum, 6 parts of prepared ginger, 6 parts of black aconite root slices, 5 parts of coptis chinensis, and 3 parts of indigo naturalis.

[0063] Secondly, the present invention provides a method for preparing a traditional Chinese medicine composition for treating ulcerative colitis. In some embodiments, the method for preparing the traditional Chinese medicine composition for treating ulcerative colitis includes: weighing raw materials according to a specified ratio, mixing all raw materials and then extracting them using conventional extraction methods, or extracting them separately using conventional extraction methods and then mixing them to obtain the final product.

[0064] In some embodiments, the conventional extraction method includes one or more of the following: decoction extraction, maceration extraction, percolation extraction, reflux extraction, ultrasonic extraction, and steam distillation extraction; the extraction is performed at least once; and each extraction takes at least 10 minutes.

[0065] And / or, the extraction solvent is selected from water or an alcohol solution with a volume percentage of 5-98%;

[0066] And / or, the weight of the extraction solvent is more than three times the total weight of the raw materials;

[0067] And / or, the temperature of the water extraction is 90-100℃.

[0068] In some embodiments, the conventional extraction method specifically includes:

[0069] (1) Weigh the raw materials according to the proportion, mix all the raw materials and add water 3-10 times the total weight of the raw materials to form a medicinal solution;

[0070] (2) Heat the medicinal liquid to 90-100℃, then keep it at 90-100℃ for 0.5-1 hour, filter, and concentrate the filtrate to obtain the product;

[0071] Optionally, in step (2), the medicinal solution is soaked for 1-2 hours before heating;

[0072] The concentration includes: concentrating the filtrate to 1 / 3-2 / 3 of its original volume at 50-70°C;

[0073] Alternatively, the conventional extraction method specifically includes:

[0074] (1) Weigh the raw materials according to the ratio, add the prepared ginger, black aconite slices, coptis and indigo to 3-10 times the weight of the raw materials in an ethanol solution with a volume percentage of 5-98%, extract at room temperature for 30-50 minutes by ultrasonic extraction, and then mix the extracts to form mixed drug solution A.

[0075] (1) Mix the charred cattail pollen, charred scutellaria baicalensis, charred angelica sinensis, charred sophora japonica and dried plum, and add water at 3-10 times the total weight of the mixed raw materials to form mixed medicine solution B;

[0076] (2) Mix the mixed medicine solution B and the mixed medicine solution A and heat to 90-100℃, then keep warm at 90-100℃ for 0.5-1 hour, filter, and concentrate the filtrate to obtain the product;

[0077] Optionally, the concentration includes concentrating the filtrate to 1 / 3-2 / 3 of its original volume at 50-70°C.

[0078] Thirdly, the present invention provides a traditional Chinese medicine preparation.

[0079] In some embodiments, the active ingredient of the traditional Chinese medicine preparation is the traditional Chinese medicine composition prepared by the method of preparing the traditional Chinese medicine composition.

[0080] Fourthly, the present invention provides the use of the traditional Chinese medicine composition, the traditional Chinese medicine composition prepared by the preparation method of the traditional Chinese medicine composition, or the traditional Chinese medicine preparation described herein in the preparation of a medicament for relieving, adjuvant treatment, or treatment of ulcerative colitis.

[0081] The traditional Chinese medicine composition of the present invention is dried using a vacuum freeze-drying agent and then pulverized into powder. The daily dosage for adults is 0.83g / kg, which can relieve, assist in the treatment or treat ulcerative colitis.

[0082] In some embodiments, the medication for relieving, adjunctive treatment or treating ulcerative colitis includes at least one of the following effects (1)-(5):

[0083] (1) Soothes the liver and strengthens the spleen, and / or astringes the intestines to stop bleeding;

[0084] (2) Inhibits intestinal inflammatory response and reduces the degree of inflammatory cell infiltration;

[0085] (3) Improves pathological and histological damage and promotes the repair of intestinal mucosal barrier function;

[0086] (4) Regulate macrophage polarization and restore the M1 / M2 macrophage balance;

[0087] (5) Regulate CD4+ T cell differentiation and reshape Th17 / Treg cell immune homeostasis.

[0088] In some embodiments, the ulcerative colitis is caused by liver yin deficiency;

[0089] And / or, the ulcerative colitis is caused by liver stagnation or liver reversal;

[0090] Optionally, the ulcerative colitis is moderate ulcerative colitis.

[0091] Liver yin deficiency is caused by excessive worry or staying up late, which damages the liver and depletes liver yin, leading to a deficiency of liver yin fluid. Liver stagnation is caused by long-term emotional distress leading to stagnation of liver qi, while liver rebellion is caused by long-term excessive liver fire leading to rebellious liver qi.

[0092] The traditional Chinese medicine composition of the present invention has a better therapeutic effect on ulcerative colitis caused by liver yin deficiency, liver stagnation, or liver reversal.

[0093] The present invention will be further described in detail below with reference to specific embodiments, which should not be construed as limiting the scope of protection claimed by the present invention.

[0094] All active pharmaceutical ingredients used in the following examples were provided by the pharmacy of Dongfang Hospital, Beijing University of Chinese Medicine. For experiments where specific experimental steps or conditions are not specified, the procedures and conditions described in the literature in this field can be followed. Unless otherwise specified, all raw materials, reagents, or instruments used are commercially available conventional reagent products.

[0095] Example 1

[0096] This embodiment provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 6g of charred sophora japonica, 8g of dried plum, 6g of prepared ginger, 6g of prepared aconite root, 5g of coptis chinensis, and 3g of indigo naturalis.

[0097] This embodiment also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, the preparation method of which is as follows:

[0098] Weigh the above ingredients according to the proportions, mix all the ingredients together, and add 10 times the total weight of water to form a mixed medicine solution.

[0099] First, soak the mixed medicinal solution for 2 hours, then heat it to 90℃, keep it at 90℃ for 0.5 hours, then filter it using a 100-mesh filter membrane, and concentrate the filtrate to 1 / 3 of its original volume at 70℃ to obtain the final product.

[0100] Example 2

[0101] This embodiment provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 4g of charred cattail pollen, 7g of charred scutellaria baicalensis, 4g of charred angelica sinensis, 7g of charred sophora japonica, 6g of dried plum, 8g of prepared ginger, 5g of prepared aconite root, 7g of coptis chinensis, and 2g of indigo naturalis.

[0102] This embodiment also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, the preparation method of which is as follows:

[0103] Weigh the above ingredients according to the proportions, mix all the ingredients together, and add water at a ratio of 5 times the total weight of the ingredients to form a mixed medicinal solution.

[0104] First, soak the mixed medicinal solution for 1.5 hours, then heat it to 95°C and keep it at 95°C for 0.5-1 hour. Then filter it using a 100-mesh filter membrane. Concentrate the filtrate to 3 / 5 of its original volume at 60°C to obtain the final product.

[0105] Example 3

[0106] This embodiment provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 6g of charred cattail pollen, 5g of charred scutellaria baicalensis, 6g of charred angelica sinensis, 5g of charred sophora japonica, 10g of dried plum, 4g of prepared ginger, 7g of prepared aconite root, 3g of coptis chinensis, and 5g of indigo naturalis.

[0107] This embodiment also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, the preparation method of which is as follows:

[0108] Weigh the above ingredients according to the proportions, mix all the ingredients together, and add water three times the total weight of the ingredients to form a mixed medicine solution.

[0109] First, soak the mixed medicinal solution for 1 hour, then heat it to 100℃, keep it at 100℃ and decoct for 1 hour, then filter it using a 100-mesh filter membrane, and concentrate the filtrate to 2 / 3 of its original volume at 50℃ to obtain the final product.

[0110] Example 4

[0111] This embodiment provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 10g of charred cattail pollen, 7g of charred scutellaria baicalensis, 6g of charred angelica sinensis, 3g of charred sophora japonica flower, 10g of dried plum, 2g of prepared ginger, 7g of prepared aconite root, 3g of coptis chinensis, and 8g of indigo naturalis.

[0112] This embodiment also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, the preparation method of which is as follows:

[0113] Weigh the above raw materials according to the ratio, add the prepared ginger, black aconite slices, coptis and indigo to 6 times the weight of the raw materials in a 50% ethanol solution, extract by ultrasonication at room temperature for 50 minutes, and then mix the extracts to form mixed drug solution A.

[0114] Mix the charred cattail pollen, charred scutellaria baicalensis, charred angelica sinensis, charred sophora japonica, and charred dried plum, then add water equal to 6 times the total weight of the mixed raw materials to form mixed medicine solution B;

[0115] Mix the medicinal solution B and the medicinal solution A, heat to 100°C, keep warm at 100°C and decoct for 1 hour, then filter using a 100-mesh filter membrane, and concentrate the filtrate to 1 / 3 of its original volume at 50°C to obtain the final product.

[0116] Example 5

[0117] This embodiment provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 2g of charred cattail pollen, 3g of charred scutellaria baicalensis, 4g of charred angelica sinensis, 8g of charred sophora japonica, 6g of dried plum, 10g of prepared ginger, 5g of black aconite root slices, 7g of coptis chinensis, and 1g of indigo naturalis.

[0118] This embodiment also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, the preparation method of which is as follows:

[0119] Weigh the above raw materials according to the ratio, add the prepared ginger, black aconite slices, coptis and indigo to 6 times the weight of the raw materials in a 50% ethanol solution, extract by ultrasonication at room temperature for 30 minutes, and then mix the extracts to form mixed drug solution A.

[0120] Mix the charred cattail pollen, charred scutellaria baicalensis, charred angelica sinensis, charred sophora japonica, and charred dried plum, then add water equal to 6 times the total weight of the mixed raw materials to form mixed medicine solution B;

[0121] Mix the medicinal solution B and the medicinal solution A, heat to 100°C, keep warm at 100°C and decoct for 1 hour, then filter using a 100-mesh filter membrane, and concentrate the filtrate to 1 / 3 of its original volume at 50°C to obtain the final product.

[0122] Comparative Example 1

[0123] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 6g of charred sophora japonica, 6g of prepared ginger, 6g of prepared aconite root, 5g of coptis chinensis, and 3g of indigo naturalis.

[0124] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0125] Comparative Example 2

[0126] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 6g of charred sophora japonica flower, 8g of dried plum, 6g of dried aconite root slices, 5g of coptis chinensis, and 3g of indigo naturalis.

[0127] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0128] Comparative Example 3

[0129] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 6g of charred sophora japonica flower, 8g of dried plum, 6g of prepared ginger, 5g of coptis chinensis, and 3g of indigo naturalis.

[0130] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0131] Comparative Example 4

[0132] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 6g of charred sophora japonica flower, 8g of dried plum, 6g of prepared ginger, 6g of prepared aconite root, 5g of coptis chinensis, and 3g of indigo naturalis.

[0133] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0134] Comparative Example 5

[0135] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 8g of dried plum, 6g of prepared ginger, 6g of prepared aconite root, 5g of coptis chinensis, and 3g of indigo naturalis.

[0136] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0137] Comparative Example 6

[0138] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 6g of Scutellaria baicalensis charcoal, 5g of Angelica sinensis charcoal, 6g of Sophora japonica charcoal, 8g of Prunus mume, 6g of Zingiber officinale (processed), 6g of Aconitum carmichaelii (processed), 5g of Coptis chinensis, and 3g of Indigo naturalis.

[0139] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0140] Comparative Example 7

[0141] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 5g of charred angelica sinensis, 6g of charred sophora japonica flower, 8g of dried plum, 6g of prepared ginger, 6g of prepared aconite root, 5g of coptis chinensis, and 3g of indigo naturalis.

[0142] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0143] Comparative Example 8

[0144] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 6g of charred sophora japonica flower, 8g of dried plum, 6g of prepared ginger, 6g of black aconite root slices, and 3g of indigo naturalis.

[0145] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0146] Comparative Example 9

[0147] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of charred cattail pollen, 6g of charred scutellaria baicalensis, 5g of charred angelica sinensis, 6g of charred sophora japonica, 8g of dried plum, 6g of prepared ginger, 6g of black aconite root slices, and 5g of coptis chinensis.

[0148] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0149] Comparative Example 10

[0150] This comparative example provides a traditional Chinese medicine composition for treating ulcerative colitis, comprising the following ingredients: 5g of Typha pollen, 6g of Scutellaria baicalensis, 5g of Angelica sinensis, 6g of Sophora japonica, 8g of Prunus mume, 6g of prepared ginger, 6g of processed aconite, 5g of Coptis chinensis, and 3g of Indigo naturalis.

[0151] This comparative example also provides a pharmaceutical preparation comprising the above-mentioned traditional Chinese medicine composition, which is prepared in the same way as in Example 1.

[0152] Experimental Example: Observation of the efficacy of the traditional Chinese medicine composition described in this invention in a mouse model of ulcerative colitis.

[0153] 1. Materials and Methods

[0154] 1.1 Laboratory Animals

[0155] 108 female, 6-week-old SPF-grade healthy C57BL / 6 mice, weighing 18-20g, were provided by Spiefol (Beijing) Biotechnology Co., Ltd. and housed in the experimental animal facility of Dongfang Hospital, Beijing University of Chinese Medicine. The mice were acclimatized for 7 days in a quiet and suitable environment, with the room temperature controlled at (22±2)℃, relative humidity maintained at 40%–65%, and 12 hours of light per day (8:00-20:00 daytime light; 20:00-8:00 nighttime darkness). During the rearing period, the mice were allowed free movement, had ample clean drinking water, and were fed standard rodent food.

[0156] 1.2 Experimental reagents:

[0157] Traditional Chinese medicine: The traditional Chinese medicine compositions of Examples 1-5 and Comparative Examples 1-10 were dried by vacuum freeze dryer and then pulverized to make medicinal powder.

[0158] Positive control drugs: Mesalazine enteric-coated granules (daily dose for mice: 606 mg / kg, drug weight / mouse body weight), prepared with distilled water to a final concentration of 606 mg / 10 ml before use, administered by gavage daily; 5% chloral hydrate (anesthetic dose: 70 μL / 10 g mouse body weight). All required drugs were provided by the pharmacy of Dongfang Hospital, Beijing University of Chinese Medicine.

[0159] 1.3 Main Reagents and Instruments

[0160] Dextran sulfate sodium salt (DSS), average molecular weight 36,000-50,000 (purchased from MP Biomedicals); fecal occult blood test strips (Zhuhai Beso Biotechnology Co., Ltd.); RPMI 1640 medium (Hyclone); FBS (fetal bovine serum, Gibco).

[0161] Percoll cell separation medium, red blood cell lysis buffer, 1×PBS buffer, penicillin and antibiotics, and bovine serum albumin were all purchased from Solarbio.

[0162] Hematoxylin staining solution, eosin staining solution, hematoxylin differentiation solution, and hematoxylin blueing solution were all purchased from Wuhan Saiweier Biotechnology Co., Ltd.

[0163] Flow cytometry antibodies CD3, CD4, F4 / 80, CD206, CD16 / 32, IL-17, and FOXP3 were all purchased from Biolegend; Trizol reagents were from Invitrogen; a DP74 microscope (Olympus); a flow cytometer (BD Biosciences); and a 7300 real-time quantitative PCR system (Applied Biosystems).

[0164] 1.4 Animal grouping, modeling, and drug administration

[0165] (1) Grouping: The experimental mice were randomly divided into the following 18 groups according to their body weight: blank group, model group, traditional Chinese medicine composition group of Examples 1-5 and Comparative Examples 1-10, and positive control group, with 6 mice in each group.

[0166] (2) Modeling: Except for the blank group mice, the other experimental groups mice were allowed to drink 2.5% (w / v) DSS solution freely for 7 days to prepare the UC model. The blank group mice were allowed to eat and drink freely throughout the process.

[0167] (3) Drug administration: After the UC model was prepared, drug administration began on day 8;

[0168] The traditional Chinese medicine compositions of Examples 1-5 or Comparative Examples 1-10 were administered the corresponding drug solutions by gavage for 7 consecutive days. The daily dose for experimental mice was 7.853 g / kg (medicinal material weight / mouse body weight). The powder obtained from the experimental drugs in 1.2 was used and stored at 4°C. Before use, the powder was prepared with distilled water to a final concentration of 7.583 mg / 10 ml. 100 μl / 10 g (drug solution / mouse body weight for each group) was administered by gavage daily.

[0169] The positive control group was given mesalazine aqueous solution (mesalazine enteric granules were prepared with distilled water to a final concentration of 606 mg / 10 ml) by gavage, 100 μL / 10 g (drug solution / mouse body weight) daily for 7 consecutive days; to maintain parallelism, other groups (blank group and model group) were given the same volume of sterile PBS at the same time.

[0170] During the intervention period, the mice were observed daily for general condition and symptoms, their weight was measured, fecal occult blood was tested, fecal characteristics were recorded, and the Disease Activity Index (DAI) was calculated. After the intervention, tissue samples were collected from the mice. Colonic tissue was collected 1 cm above the anus, pathologically embedded, and stained with hematoxylin and eosin (HE) to assess the degree of tissue damage in mice with ulcerative colitis. Assessment indicators included the number of crypts, the extent of intestinal epithelial damage, and the extent of mucosal and submucosal damage. Spleen samples were ground using a 70µm cell sieve, subjected to schizochrome staining, density gradient centrifugation, and flow cytometry staining. Flow cytometry was used to assess the content of major macrophage subsets in the mouse spleen. Mesenteric lymph node samples were ground, subjected to density gradient centrifugation, and flow cytometry staining. Flow cytometry was used to assess the content of major CD4+ T lymphocyte subsets in the mouse mesenteric lymph nodes.

[0171] 1.5 Observation Indicators and Methods

[0172] 1.5.1 General Observation

[0173] During the experiment, the mice's weight, diet, activity level, mental state, fur condition, frequency and characteristics of defecation were observed and recorded.

[0174] 1.5.2 Efficacy Evaluation Criteria

[0175] The Disease Activity Index (DAI) was calculated based on the criteria for ulcerative colitis mice (UC mice). DAI = (weight loss index + fecal characteristics + fecal occult blood) / 3, and the specific evaluation criteria are shown in Table 1.

[0176] Table 1 Scoring criteria for the Disease Activity Index

[0177]

[0178] 1.5.3 Colon length and spleen-to-body weight ratio

[0179] When collecting samples, the total length of the mouse colon was measured, the spleen was removed and weighed, and the spleen-to-body weight ratio was calculated.

[0180] 1.5.4 Pathological morphological observation

[0181] During tissue collection, a transverse section of the mouse colon, 1-1.5 cm from the anus, was harvested. Feces were removed, and the tissue was washed with PBS and fixed in 10% formalin for 24 hours. After removal, the tissue was dehydrated in a gradient of alcohols, then embedded in paraffin, sectioned at 5 μm thickness, mounted, dewaxed, stained with hematoxylin and eosin (HE), and mounted for observation of the mouse colon tissue morphology and structure under a light microscope. During the preparation process, it is crucial to fix the animal tissue rapidly after sacrifice to preserve its original morphological structure. The scalpels and scissors used for cutting tissue blocks must be sharp, and the tissue should not be gripped too tightly to avoid deformation of the tissue and cells due to compression. Tissue should be accurately collected according to the anatomical location, and the material should be kept clean.

[0182] 1.5.5 Flow cytometry detection of macrophage subsets in the spleen of UC mice

[0183] The entire procedure was performed at 4°C. Mouse spleens were harvested, and the tissue was gently crushed using a sterile syringe plunger. The cells were filtered through a 70μm cell sieve, subjected to erythrolysis for 10 min, centrifuged, the supernatant was discarded, and the cells were resuspended and blocked for 20 min. After centrifugation at 1600 rpm for 5 min at 4°C, the cells were resuspended. Flow cytometry antibodies for F4 / 80, CD16 / 32, and CD206 were added, and the cells were incubated at 4°C in the dark for 40 min for cell membrane staining. After washing and centrifugation, the cells were resuspended and analyzed by flow cytometry. Flowjo was used to analyze the flow cytometry data.

[0184] 1.5.6 Flow cytometry detection of CD4+ T cell subsets in mesenteric lymph nodes of UC mice

[0185] The entire procedure was performed at 4°C. Mesenteric lymph nodes from mice were collected, and the tissue was gently crushed using a sterile syringe plunger. The mixture was filtered through a 70μm cell sieve to obtain a single-cell suspension of mesenteric lymph nodes. After centrifugation and discarding the supernatant, the cells were resuspended and blocked for 20 min. The cells were then centrifuged again at 4°C, 1600 rpm for 5 min and resuspended. Stimulants were added, and the cells were incubated at 37°C with 5% CO2 for 8 h to stimulate the secretion of related factors. After stimulation, the cells were collected, centrifuged, and the supernatant was discarded. After blocking for 20 min, the cells were centrifuged again and the supernatant was discarded. CD3 and CD4 flow cytometry antibodies were added for surface antibody staining, and the cells were incubated at 4°C in the dark for 40 min for cell membrane staining. The cells were fixed with fixative, and after rupture of the membrane for 40 min, IL-17 and FOXP3 flow cytometry antibodies were added for staining. The cells were incubated at 4°C in the dark for 40 min for staining. After washing and centrifugation, the cells were resuspended and analyzed by flow cytometry. Flowjo was used to analyze the flow cytometry data.

[0186] 2. Data Statistics

[0187] SPSS 22.0 and GraphPad Prism 9.5 software were used for data analysis and graphing. Quantitative data were... The data were tested for homogeneity of variance, and one-way ANOVA was used for comparisons between groups. When the variances were homogeneous, the LSD test was used for comparisons between the two groups, and the Dunnett t3 test was used for comparisons between groups with unequal variances. The overall efficacy rate was categorical data, and the chi-square test was used. A p-value < 0.05 was considered statistically significant.

[0188] 3 Results

[0189] 3.1 General Information for Each Group

[0190] All groups of mice tolerated the modeling and drug intervention well. Behavioral performance was observed in each group. The control group showed no abnormalities, was energetic, had a good appetite and was active, with glossy and smooth fur. Compared to the control group, the UC mice induced by DSS exhibited obvious intestinal symptoms, such as varying degrees of bloody stools, diarrhea, and weight loss, as well as lethargy, slow movement, poor appetite, dull fur, arched backs, and gregarious behavior. After 7 days of administration of the herbal compositions or positive control solutions of Examples 1-5 or Comparative Examples 1-10, the mice showed significantly reduced mental state, behavioral changes, dull fur, diarrhea, and bloody stools compared to the model group, with Examples 1-5 showing better improvement than Comparative Examples 1-10.

[0191] 3.2 Changes in body weight of mice in each group

[0192] As seen Figure 1 As shown, the body weight of mice in the blank control group did not change significantly. Compared with the blank control group, the body weight of mice in the model group decreased significantly starting from day 3 after modeling, and gradually increased from day 8 onwards (P < 0.05). Compared with the model group, the body weight of mice in Examples 1-5, Comparative Examples 1-10, and the positive control group also increased significantly from day 8 to 14, and the increasing trend was greater than that in the model group (Examples 1-5). * P < 0.05 ** P < 0.01, positive control group # P < 0.05 (P > 0.05 for comparative groups 1-10); the weight recovery trend of comparative groups 1-10 during the experiment was weaker than that of mice in groups 1-5 of Examples.

[0193] 3.3 DAI score results of mice in each group

[0194] like Figure 2 As shown, the DAI scores of mice in the blank control group remained essentially unchanged; compared with the blank control group, the DAI scores of mice in the model group, Examples 1-5 groups, and Comparative Examples 1-10 groups were significantly increased from day 7 after modeling (P < 0.05), and gradually decreased from day 8 onwards; compared with the model group, the DAI scores of mice in Examples 1-5 groups ( * P < 0.05), comparative groups 1-10 (P > 0.05), positive control group ( #(P < 0.05) The DAI scores of mice also began to decrease, and the decreasing trend was greater than that of the model group. The DAI decreasing trend of comparative groups 1-10 during the experiment was weaker than that of mice in Example 1-5.

[0195] 3.4 Colon length of mice in each group

[0196] like Figure 3 As shown, compared with the blank group, the colon length in the model group was significantly shortened. ## P < 0.01), colonic rigidity and obvious intestinal fibrosis were observed; compared with the model group, after treatment, the colon length of groups 1-5, comparative groups 1-10, and the positive control group was significantly restored (Groups 1-5). * P < 0.05 ** P < 0.01, comparative examples 1-10 * P < 0.05 ** P < 0.01, positive control group ** (P < 0.01), among which the colon length of mice in Examples 1-5 tended to be higher than that in the positive control group. The degree of recovery of colon length in Comparative Examples 1-10 was lower than that in Examples 1-5. Table 2, Figure 3 The colon length of UC mice in each group is shown.

[0197] Table 2 Colon length of mice in each group

[0198]

[0199] 3.5 Spleen and body weight ratio of mice in each group

[0200] like Figure 4 As shown, the spleen in the control group was normal. Compared with the control group, the spleen size and length in the model group were significantly increased, and the spleen-to-body weight ratio was significantly higher. ## P < 0.01); Compared with the model group, after treatment, the spleen size and length in Examples 1-5, Comparative Examples 1-10, and the positive control group showed a trend of recovery, and the spleen weight was significantly lower than that in the model group (Example 1-5 groups). * P < 0.05 ** P < 0.01, P > 0.05 for comparative groups 1-10, and for the positive control group. ** (P < 0.01), among which the spleen-to-body weight ratio of UC mice in Examples 1-5 tended to be lower than that in the positive control group. The decreasing trend in Comparative Examples 1-10 was weaker than that in Examples 1-5 mice. Table 3. Figure 4 The spleen weight ratio of UC mice in each group.

[0201] Table 3 Spleen-to-body weight ratio of mice in each group

[0202]

[0203] 3.6 HE pathological staining and scoring

[0204] Under a light microscope, as Figure 5 As shown, the control group showed no edema or inflammatory response. Compared with the control group, the model group showed significant edema of the colonic mucosa, intestinal epithelial shedding, loss of villi, glandular deformation, and local erosion, bleeding, and inflammatory cell infiltration. ## P < 0.01. Compared with the model group, after treatment, the glandular structure of mice in Examples 1-5, Comparative Examples 1-10, and the positive control group showed significant recovery, reduced edema, and improved inflammatory cell infiltration, with no obvious erosion or epidermal cell shedding. * P<0.05 ** P < 0.01); the degree of recovery of mucosa and glands and reduction of edema in comparative groups 1-10 were also reduced (P > 0.05); the positive control group showed a certain degree of recovery of glandular structure, reduced edema compared with the model group, improved inflammatory cell infiltration, and reduced erosion. * (P<0.05); Among them, the degree of recovery of mucosa and glands and reduction of edema in groups 1-5 of Examples were slightly better than those in the positive control group (HE staining, ×200). The degree of recovery of mucosa and glands and reduction of edema in groups 1-10 of Comparative Examples were also reduced, but the degree of recovery and reduction was weaker than that in groups 1-5 of Examples.

[0205] Table 4 Figure 5 HE staining and pathological scoring of UC mice in each group; Figure 5 These are the HE staining and pathological scores of mice in each group in the experimental examples of this invention; where A: HE staining of mice in each group (×200); B: pathological histological scores of mice in each group.

[0206] Table 4. Histopathological scores of mice in each group

[0207]

[0208] 3.7 Regulatory effects of macrophages in different groups of mice

[0209] Mouse spleens were collected, single-cell suspensions were prepared for staining, and the results were analyzed using flow cytometry. Figure 6 Compared with the model group, the content of M1 macrophages (F4 / 80+, CD16 / 32+) in mice in Examples 1-5, Comparative Examples 1-10, and the positive control group was significantly reduced (Example 1-5 groups). * P < 0.05, P > 0.05 for comparative examples 1-10, and P > 0.05 for the positive control group. *P < 0.05), with the decreasing trend in Examples 1-5 groups being higher than that in Comparative Examples 1-10 groups; compared with the model group, the content of M2 macrophages (F4 / 80+, CD206+) in UC mice in Examples 1-5 groups, Comparative Examples 1-10 groups, and the positive control group was significantly increased (Example 1-5 groups). ** P < 0.01, P > 0.05 for comparative groups 1-10, and for the positive control group. * P < 0.05), where the increasing trend of Examples 1-5 was higher than that of Comparative Examples 1-10.

[0210] Figure 6 and Figure 7 Comparison of the regulatory effects of splenic macrophages in different groups of UC mice;

[0211] Figure 6 Comparison of the regulatory effects of M1 macrophages in different groups of UC mice; where A: M1 macrophages in UC mice; B: Analysis of the content level of M1 macrophages in UC mice;

[0212] Figure 7 To compare the regulatory effects of M2 macrophages in different groups of UC mice; where A: M2 macrophages in UC mice; B: Analysis of the content level of M2 macrophages in UC mice.

[0213] 3.8 CD4+ T cells in the mesenteric lymph nodes of mice in each group

[0214] Mesenteric lymph nodes from mice were collected, single-cell suspensions were prepared for staining, and the results were analyzed using flow cytometry. Figure 7 As shown, compared with the model group, the levels of Th17 cells (CD4+, IL-17+) in mice in Examples 1-5, Comparative Examples 1-10, and the positive control group were significantly decreased (Example 1-5 groups). * P < 0.05, P > 0.05 for comparative examples 1-10, and P > 0.05 for the positive control group. * P < 0.05), with the decreasing trend in Examples 1-5 groups being greater than that in Comparative Examples 1-10 groups; compared with the model group, the content of Treg cells (CD4+, FOXP3+) in mice in Examples 1-5 groups, Comparative Examples 1-10 groups, and the positive control group was significantly increased (Example 1-5 groups). ** P < 0.01, P > 0.05 for comparative groups 1-10, and for the positive control group. * P < 0.05), where it can be seen that the Treg cell content in the Example 1-5 groups tends to be higher than that in the positive control group, and the increasing trend in the Comparative Example 1-10 groups is slower than that in the Example 1-5 groups.

[0215] Figure 8 and Figure 9 The regulation of CD4+ T cells in the mesenteric lymph nodes of UC mice in each group;

[0216] Figure 8 The regulation of Th17 cells in UC mice in each group; where A: Th17 cells in UC mice; B: Th17 cell content level analysis in UC mice.

[0217] Figure 9 The regulation of Treg cells in UC mice in each group; A: Treg cells in UC mice; B: Analysis of Treg cell content levels in UC mice.

[0218] The above experimental examples demonstrate that the traditional Chinese medicine composition of this invention can inhibit intestinal inflammatory response in mice with ulcerative colitis, reduce the degree of inflammatory cell infiltration, improve pathological histological damage, promote intestinal mucosal repair, regulate macrophage polarization, restore the balance of M1 / M2 macrophages, regulate CD4+ T cell differentiation, and reshape Th17 / Treg cell immune homeostasis, providing new ideas and strategies for targeted immunotherapy of ulcerative colitis.

[0219] Obviously, the above embodiments are merely illustrative examples for clear explanation and are not intended to limit the implementation. Those skilled in the art will recognize that other variations or modifications can be made based on the above description. It is neither necessary nor possible to exhaustively list all possible implementations here. However, obvious variations or modifications derived therefrom are still within the scope of protection of this invention.

Claims

1. A traditional Chinese medicine composition for treating moderate to severe ulcerative colitis, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 2-10 parts of charred cattail pollen, 3-7 parts of charred scutellaria baicalensis, 4-6 parts of charred angelica sinensis, 3-8 parts of charred sophora japonica, 6-10 parts of dried plum, 2-10 parts of prepared ginger, 5-7 parts of black aconite, 3-7 parts of coptis chinensis, and 1-8 parts of indigo naturalis.

2. The traditional Chinese medicine composition for treating ulcerative colitis according to claim 1, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 4-6 parts of charred cattail pollen, 5-7 parts of charred scutellaria baicalensis, 4-6 parts of charred angelica sinensis, 5-7 parts of charred sophora japonica, 6-10 parts of dried plum, 4-8 parts of prepared ginger, 5-7 parts of black aconite, 3-7 parts of coptis chinensis, and 2-5 parts of indigo naturalis.

3. The traditional Chinese medicine composition for treating ulcerative colitis according to claim 2, characterized in that, The traditional Chinese medicine composition is made from the following raw materials in parts by weight: 5 parts of charred cattail pollen, 6 parts of charred scutellaria baicalensis, 5 parts of charred angelica sinensis, 6 parts of charred sophora japonica, 8 parts of dried plum, 6 parts of prepared ginger, 6 parts of black aconite, 5 parts of coptis chinensis, and 3 parts of indigo naturalis.

4. A method for preparing a traditional Chinese medicine composition for treating ulcerative colitis according to any one of claims 1-3, characterized in that, include: Weigh the raw materials according to the specified ratio, mix all the raw materials and then extract them using conventional extraction methods, or extract them separately using conventional extraction methods and then mix them to obtain the final product.

5. The preparation method according to claim 4, characterized in that, The conventional extraction method includes one or more of the following: decoction extraction, maceration extraction, percolation extraction, reflux extraction, ultrasonic extraction, and steam distillation extraction; the extraction is performed at least once; and each extraction takes at least 10 minutes. And / or, the extraction solvent is selected from water or an alcohol solution with a volume percentage of 5-98%; And / or, the weight of the extraction solvent is more than three times the total weight of the raw materials; And / or, the temperature of the water extraction is 90-100℃.

6. The preparation method according to claim 5, characterized in that, The conventional extraction method specifically includes: (1) Weigh the raw materials according to the proportion, mix all the raw materials and add water at 3-10 times the total weight of the raw materials to form a medicinal solution; (2) Heat the medicinal liquid to 90-100℃, then keep it at 90-100℃ for 0.5-1 hour, filter, and concentrate the filtrate to obtain the product.

7. The preparation method according to claim 6, characterized in that, In step (2), the medicinal solution is soaked for 1-2 hours before heating; The concentration includes: concentrating the filtrate to 1 / 3-2 / 3 of its original volume at 50-70°C; Alternatively, the conventional extraction method specifically includes: (1) Weigh the raw materials according to the ratio, add the prepared ginger, black aconite slices, coptis and indigo to 3-10 times the weight of the raw materials in an ethanol solution with a volume percentage of 5-98%, extract at room temperature for 30-50 minutes by ultrasonic extraction, and then mix the extracts to form mixed drug solution A. (1) Mix the charred cattail pollen, charred scutellaria baicalensis, charred angelica sinensis, charred sophora japonica and dried plum, and add water at 3-10 times the total weight of the mixed raw materials to form mixed medicine solution B; (2) Mix the mixed medicine solution B and the mixed medicine solution A and heat to 90-100℃. Then keep warm and decoct at 90-100℃ for 0.5-1 hour, filter, and concentrate the filtrate to obtain the product.

8. The preparation method according to claim 7, characterized in that, The concentration includes concentrating the filtrate to 1 / 3-2 / 3 of its original volume at 50-70°C.

9. A traditional Chinese medicine preparation, characterized in that, The traditional Chinese medicine preparation is prepared by the method of preparing the traditional Chinese medicine composition according to any one of claims 1-3 or the traditional Chinese medicine composition according to any one of claims 4-8, with the traditional Chinese medicine composition as the active ingredient. The traditional Chinese medicine preparation also includes pharmaceutically acceptable excipients.

10. The traditional Chinese medicine preparation according to claim 9, characterized in that, The traditional Chinese medicine preparations are in the form of liquid or solid preparations.

11. The traditional Chinese medicine preparation according to claim 10, characterized in that, The dosage forms of the traditional Chinese medicine preparations are tablets, capsules, powders, granules, or ointments.

12. Use of the traditional Chinese medicine composition according to any one of claims 1-3, the traditional Chinese medicine composition prepared by the preparation method of any one of claims 4-8, or the traditional Chinese medicine preparation according to any one of claims 9-11 in the preparation of a medicament for relieving, adjuvant treatment, or treatment of ulcerative colitis.

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