Traditional Chinese medicine preparation for treating ulcerative colitis and application thereof

Abstract

The invention provides a traditional Chinese medicine preparation for treating ulcerative colitis and application thereof, and relates to the technical field of traditional Chinese medicine pharmacy, and the traditional Chinese medicine preparation is prepared from the following eleven traditional Chinese medicines: radix aconiti lateralis preparata, herba epimedii, herba cistanche, ginger processed cortex magnoliae officinalis, rhizoma cyperi, acanthopanax, herba lycopi, humifuse euphorbia herb, fructus chebulae, cuttlebone and honey-fried licorice root. The traditional Chinese medicine preparation disclosed by the invention can be used for effectively inhibiting oxidative stress injury of H2O2 induced IEC-6 cells; meanwhile, the level of inflammatory factors TNF-alpha and IL-1beta secreted by IEC-6 cells induced by LPS can be remarkably reduced, so that inflammatory response is relieved; besides, the traditional Chinese medicine preparation can obviously reduce the DAI score of an ulcerative colitis model mouse, repair and improve the structural integrity of colon tissue, enhance the intestinal barrier function, inhibit the expression of proinflammatory cytokines IL-6, TNF-alpha and IL-1beta, promote the expression of an anti-inflammatory cytokine IL-10 and effectively improve the inflammatory reaction, has a remarkable anti-oxidation effect, and can be used for preparing the traditional Chinese medicine preparation for treating the ulcerative colitis. A new treatment scheme is provided for the treatment of ulcerative colitis.

Description

Technical Field

[0001] The invention relates to the technical field of traditional Chinese medicine pharmaceutical preparation, and in particular to a traditional Chinese medicine preparation for treating ulcerative colitis and application thereof. Background Art

[0002] Ulcerative colitis (UC) usually develops from distal to proximal. It is a chronic disease that affects the rectum and colon, manifested by persistent inflammatory response and ulcer formation. Clinical manifestations include diarrhea, abdominal pain and cramps before and after defecation, urgent defecation, mucus, pus and blood in stool, fever and weight loss, and there may be repeated attacks and remissions. Its etiology is not yet fully understood. Risk factors include but are not limited to recent bacterial infection, family history of UC, diet, immune system abnormalities and environmental factors. With the changes in modern lifestyle and environment, the incidence rate has gradually increased. The possibility of patients developing colorectal cancer in the late stage of the disease is 70% higher than that of the general population, which to some extent reduces the health level and quality of life of patients. At present, the drug treatment for ulcerative colitis is mainly aminosalicylic acid, glucocorticoids and immunosuppressants, but there are shortcomings such as difficulty in treatment, long medication cycle, easy recurrence and many adverse reactions, which limit its clinical application.

[0003] In traditional Chinese medicine theory, UC is often classified as "intestinal problems, diarrhea, chronic dysentery" and other categories. Its basic pathogenesis is spleen deficiency and dampness, intestinal dysfunction, and the site of disease is the spleen, stomach and large intestine, which is closely related to the liver and kidney. "Medical Enlightenment" records: "The method of treating dysentery must be combined with the meaning of treating carbuncle", which can be attributed to "internal ulcer". The "redness, swelling, heat and pain" of inflammation is similar to the pathogenic characteristics of heat evil "burning and inflammation, easy to cause sores and carbuncle" to a certain extent; the typical symptoms of UC are diarrhea, mucus, pus and blood in the stool, and the pathogenic characteristics of dampness "sticky and downward" are similar to the exudation of inflammatory reaction. Studies have found that some traditional prescriptions and Chinese medicine extracts can effectively relieve inflammation and clinical symptoms of UC animal models by regulating the function of intestinal mucosal barrier. In addition, compared with Western medical treatment, Chinese medicine focuses on understanding the disease from a holistic perspective and treating it based on syndrome differentiation. It has significant efficacy in treating ulcerative colitis, alleviating patient symptoms, and reducing disease recurrence, and has the advantages of low price, low recurrence, and fewer adverse reactions. However, the current treatment of ulcerative colitis with traditional Chinese medicine still has disadvantages such as relatively slow efficacy and low acceptance among some patients. Therefore, there is an urgent need to develop an efficient, safe and easily accepted Chinese medicine preparation to better meet the treatment needs of patients with ulcerative colitis. Summary of the invention

[0004] 1. Technical issues to be solved

[0005] In view of the deficiencies in the prior art, the present invention provides a traditional Chinese medicine preparation for treating ulcerative colitis and application thereof.

[0006] (II) Technical solution

[0007] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0008] The invention provides a traditional Chinese medicine preparation for treating ulcerative colitis. The traditional Chinese medicine preparation is composed of the following raw materials in weight fractions: 10-28 parts of processed aconite root, 8-26 parts of epimedium, 3-22 parts of cistanche deserticola, 12-30 parts of zingiber officinale, 5-28 parts of cyperus rotundus, 5-27 parts of acanthopanax senticosus, 5-27 parts of zedoaria, 8-27 parts of euphorbia cerevisiae, 10-32 parts of terminalia chebula, 3-20 parts of cuttlebone and 10-32 parts of roasted liquorice.

[0009] Furthermore, the Chinese medicine preparation is composed of the following raw materials in weight fractions: 15-25 parts of processed aconite, 10-20 parts of epimedium, 5-15 parts of Cistanche deserticola, 15-25 parts of Magnolia officinalis, 10-20 parts of Cyperus rotundus, 10-20 parts of Acanthopanax senticosus, 10-20 parts of Eupatorium fortunei, 10-20 parts of Euphorbia milii, 10-20 parts of Herba Euphorbiae, 15-25 parts of Terminalia chebula, 5-15 parts of Cuttlebone, and 15-25 parts of Radix Glycyrrhizae.

[0010] Furthermore, the Chinese medicine preparation is composed of the following raw materials in weight fractions: 20 parts of processed aconite, 15 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of Magnolia officinalis, 15 parts of Cyperus rotundus, 15 parts of Acanthopanax senticosus, 15 parts of Eupatorium fortunei, 15 parts of Herba Euphorbiae, 20 parts of Terminalia chebula, 10 parts of Cuttlebone, and 20 parts of Radix Glycyrrhizae.

[0011] The present invention provides a method for preparing a Chinese medicine preparation for treating ulcerative colitis. The Chinese medicine preparation is prepared into any oral dosage form acceptable to the human body, including pills, powders, granules, oral liquids, decoctions and tablets, by conventional pharmaceutical methods and with medically acceptable adjuvants.

[0012] Furthermore, a Chinese medicine decoction for ulcerative colitis is prepared according to the following steps:

[0013] (1) Prepare aconite root, magnolia bark and licorice as follows: Prepared aconite root: soak aconite root in a bile solution overnight, then add salt and continue soaking, take out and dry in the sun every day, and gradually extend the drying time until a large number of crystallized salt particles (salt frost) appear on the surface of the aconite root and the body becomes hard, thus obtaining the prepared aconite root; Ginger magnolia bark: wash and mash ginger, add appropriate amount of water, squeeze out ginger juice, take magnolia bark, add ginger juice in a ratio of 10:1, mix well, put in a pot and stir-fry over low heat until the ginger juice is absorbed, take out and dry to obtain ginger magnolia bark; Roasted licorice root: dilute refined honey with appropriate amount of boiling water, add licorice in a ratio of 4:1, mix well, simmer thoroughly, put in a pot and stir-fry over low heat until it turns yellow to dark yellow, take out and cool when it is no longer sticky, thus obtaining roasted licorice root.

[0014] (2) Weigh each raw material according to the above weight proportions, wash, dry, and grind into coarse powder, and sieve through a 24-mesh sieve.

[0015] (3) Put the above coarse powder in a pot, add water to submerge the coarse powder, soak for 20-30 minutes, heat to boiling, keep slightly boiling for 30-60 minutes, filter the decoction, boil the residue again 1-2 times according to the above method, combine the filtrate, and concentrate under reduced pressure and vacuum to a drug mass concentration of 1 g / mL to obtain the water decoction of the Chinese medicine preparation of the present invention.

[0016] The present invention provides application of the traditional Chinese medicine preparation in preparing medicine for preventing and / or treating ulcerative colitis.

[0017] The pharmacological effects of each Chinese medicine component in the Chinese medicine preparation of the present invention are as follows:

[0018] Processed Aconite: Spicy and sweet, hot in nature, enters the heart, kidney, and spleen meridians; has the effects of restoring yang, replenishing fire and supporting yang, dispelling cold and relieving pain; used for yang-deficiency collapse, cold limbs and weak pulse, insufficient heart yang, chest pain, vomiting and diarrhea due to deficiency and cold, cold pain in the abdomen, kidney yang deficiency, impotence and cold uterus, edema due to yin cold, exogenous infection due to yang deficiency, and pain due to cold and dampness. Aconite contains a large amount of alkaloid components, which have strong toxic effects, so it is often processed in clinical practice. As its main active ingredient, alkaloids also show good anti-inflammatory effects in the treatment of UC.

[0019] Epimedium: spicy, sweet, warm in nature; enters the liver and kidney meridians; has the effects of tonifying kidney yang, strengthening tendons and bones, and removing rheumatism; used for kidney yang deficiency, impotence and spermatorrhea, tendon and bone weakness, rheumatic arthralgia, numbness and cramps.

[0020] Cistanche deserticola: sweet and salty in taste, warm in nature, enters the kidney and large intestine meridians; has the effects of tonifying kidney yang, replenishing essence and blood, and moistening the intestines and promoting bowel movements; used for insufficient kidney yang, deficiency of essence and blood, impotence and infertility, soreness of the waist and knees, weakness of tendons and bones, and dry intestines and constipation.

[0021] Ginger Magnolia Bark: bitter, pungent, warm in nature; enters the spleen, stomach, lung, and large intestine meridians; has the effects of drying dampness and eliminating phlegm, and removing fullness from the lower qi; used for dampness stagnation, abdominal distension, vomiting and diarrhea, food accumulation and qi stagnation, abdominal distension and constipation, phlegm and cough. Roasting ginger can eliminate its irritation to the throat and enhance the function of warming the middle and harmonizing the stomach.

[0022] Cyperus rotundus: spicy, slightly bitter, slightly sweet, neutral in nature; enters the liver, spleen, and triple burner meridians; has the effects of soothing the liver and relieving depression, regulating qi and relieving fullness, regulating menstruation and relieving pain; used for liver depression and qi stagnation, chest and flank distension and pain, hernia pain, breast distension and pain, spleen and stomach qi stagnation, abdominal distension and pain, irregular menstruation, amenorrhea and dysmenorrhea.

[0023] Acanthopanax: spicy, slightly bitter, warm in nature, enters the spleen, kidney, and heart meridians; has the effects of invigorating qi and strengthening the spleen, nourishing the kidney and calming the mind; used for spleen and lung qi deficiency, physical weakness, loss of appetite, lung and kidney deficiency, long-term cough and asthma, kidney deficiency, low back and knee pain, heart and spleen deficiency, insomnia and dreaminess.

[0024] Ze Lan: bitter, pungent, slightly warm in nature. Enters the liver and spleen meridians; has the effects of promoting blood circulation and regulating menstruation, removing blood stasis and eliminating carbuncle, promoting diuresis and reducing swelling; used for irregular menstruation, amenorrhea, dysmenorrhea, abdominal pain caused by blood stasis after childbirth, swelling and poison caused by sores, carbuncle, edema and ascites.

[0025] Herba Dijincao: spicy in taste, neutral in nature; enters the liver and large intestine meridians; has the effects of clearing away heat and detoxifying, cooling blood and stopping bleeding, promoting dampness and relieving jaundice; used for dysentery, diarrhea, hemoptysis, hematuria, bloody stool, metrorrhagia, sores, carbuncles, and damp-heat jaundice.

[0026] Terminalia chebula: bitter, sour, and astringent in taste, neutral in nature, enters the lung and large intestine meridians; has the effects of astringing the intestines to stop diarrhea, astringing the lungs to stop coughing, reducing fire and relieving sore throat; used for long-term diarrhea and dysentery, rectal prolapse due to blood in the stool, lung deficiency, wheezing and coughing, long-term cough, sore throat and hoarseness.

[0027] Cuttlebone: salty and astringent in taste, warm in nature, enters the spleen and kidney meridians; has the effects of astringing and stopping bleeding, astringing semen and stopping leukorrhea, relieving acid and relieving pain, and astringing moisture and healing ulcers; used for vomiting blood, epistaxis, metrorrhagia, bloody stool, spermatorrhea, leukorrhea, stomach pain and acid regurgitation; used externally for bleeding injuries, eczema, eczema, and ulcers that do not heal.

[0028] Roasted licorice root: sweet in taste, neutral in nature, enters the heart, lung, spleen, and stomach meridians; has the effects of tonifying the spleen and stomach, replenishing qi and restoring the pulse; used for weak spleen and stomach, fatigue, palpitations, and irregular pulse.

[0029] (III) Beneficial effects

[0030] The invention provides a traditional Chinese medicine preparation for treating ulcerative colitis and application thereof. The traditional Chinese medicine preparation is composed of eleven traditional Chinese medicines, namely, processed aconite root, epimedium, cistanche deserticola, ginger magnolia bark, cyperus rotundus, acanthopanax senticosus, zedoaria, herba rutaecarpae, terminalia chebula, cuttlebone and roasted licorice. The processed aconite root is used as the main medicine in the prescription, which can warm the middle and dispel cold, replenish fire and benefit earth, and has the effects of restoring yang and rescuing adverse conditions, replenishing fire and yang, dispelling cold and relieving pain; the epimedium, cistanche deserticola, ginger magnolia bark and cyperus rotundus are used as the assistant medicines, the epimedium can enhance the power of the processed aconite root in warming and tonifying kidney yang, the cistanche deserticola has mild medicinal properties, and can synergistically enhance the effects of tonifying kidney yang and benefiting essence and blood, and the combination of cyperus rotundus and ginger magnolia bark can enhance the power of promoting qi, so that the whole body qi can be unobstructed, and the stagnation of kidney-tonifying medicine can be prevented. Qi movement, which helps the transportation and absorption of drugs; Acanthopanax, Zedoaria, Herba Dijincao, Terminalia chebula, and Cuttlebone are used as adjuvants. Acanthopanax can not only assist the main and auxiliary drugs in tonifying the kidney, but also take into account the spleen and stomach, so that the acquired foundation can be nourished and the innate kidney can be better nourished; Zedoaria and Herba Dijincao can promote blood circulation and stop bleeding, which can prevent the warming yang drugs from helping fire to generate heat while preventing the occurrence of endogenous blood stasis and damp heat or bleeding. Terminalia chebula and Cuttlebone can astringe the intestines and stop diarrhea, and are used together with kidney-yang tonifying drugs to warm and astringe, treat both the symptoms and the root causes. At the same time, Cuttlebone can protect the spleen and stomach to prevent the drugs from irritating the gastrointestinal tract; Roasted Licorice is used as a guiding drug to tonify the spleen and replenish Qi while harmonizing the various drugs, so that the whole prescription can attack and tonify at the same time, warm without dryness, and tonify without stagnation.

[0031] The Chinese medicine preparation of the present invention can effectively inhibit the oxidative stress damage of IEC-6 cells induced by H2O2; at the same time, it can also significantly reduce the levels of inflammatory factors TNF-α and IL-1β secreted by IEC-6 cells induced by LPS, thereby alleviating the inflammatory response; in addition, the Chinese medicine preparation of the present invention can significantly reduce the DAI score of ulcerative colitis model mice, repair and improve the structural integrity of colon tissue, enhance the intestinal barrier function, inhibit the expression of pro-inflammatory cytokines IL-6, TNF-α and IL-1β, promote the expression of anti-inflammatory cytokine IL-10, effectively improve the inflammatory response, and has a significant antioxidant effect, thereby providing a new treatment plan for the treatment of ulcerative colitis. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The effect of each group of drugs on the colon length of UC mice. Note: A is the Control group, B is the Model group, C is the Etiasa group, D is the TL group, E is the TM dosage group, and F is the TH dosage group.

[0033] Figure 2 The effect of each group of drugs on HE staining of colon tissue of UC mice (×200). DETAILED DESCRIPTION

[0034] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0035] Example 1

[0036] A traditional Chinese medicine preparation for treating ulcerative colitis and application thereof. The traditional Chinese medicine preparation is composed of the following raw materials in weight fractions: 20 parts of processed aconite root, 15 parts of epimedium, 10 parts of cistanche deserticola, 20 parts of zingiber officinale, 15 parts of cyperus rotundus, 15 parts of acanthopanax senticosus, 15 parts of zedoaria, 15 parts of euphorbia pulegosae, 20 parts of terminalia chebula, 10 parts of cuttlebone and 20 parts of roasted liquorice.

[0037] Example 2

[0038] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 28 parts of processed aconite, 26 parts of epimedium, 22 parts of Cistanche deserticola, 30 parts of Magnolia officinalis, 28 parts of Cyperus rotundus, 27 parts of Acanthopanax senticosus, 27 parts of Zedoariae, 27 parts of Herba Euphorbiae, 32 parts of Terminalia chebula, 20 parts of Cuttlebone, and 32 parts of Radix Glycyrrhizae.

[0039] Example 3

[0040] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 10 parts of processed aconite, 8 parts of epimedium, 3 parts of Cistanche deserticola, 12 parts of Magnolia officinalis, 5 parts of Cyperus rotundus, 5 parts of Acanthopanax senticosus, 5 parts of Zedoariae, 8 parts of Herba Euphorbiae, 10 parts of Terminalia chebula, 3 parts of Cuttlebone, and 10 parts of Radix Glycyrrhizae Preparata.

[0041] Example 4

[0042] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 27 parts of processed aconite, 24 parts of epimedium, 20 parts of Cistanche deserticola, 28 parts of Magnolia officinalis, 25 parts of Cyperus rotundus, 25 parts of Acanthopanax senticosus, 26 parts of Zedoariae, 25 parts of Herba Euphorbiae, 30 parts of Terminalia chebula, 18 parts of Cuttlebone, and 30 parts of Radix Glycyrrhizae Preparata.

[0043] Example 5

[0044] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 15 parts of processed aconite, 18 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of Magnolia officinalis, 14 parts of Cyperus rotundus, 12 parts of Acanthopanax senticosus, 12 parts of Zedoariae, 16 parts of Herba Euphorbiae, 18 parts of Terminalia chebula, 8 parts of Cuttlebone, and 15 parts of Radix Glycyrrhizae Preparata.

[0045] Example 6

[0046] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 12 parts of processed aconite, 10 parts of epimedium, 5 parts of Cistanche deserticola, 14 parts of Magnolia officinalis, 7 parts of Cyperus rotundus, 7 parts of Acanthopanax senticosus, 7 parts of Zedoariae rhizome, 10 parts of Herba Euphorbiae, 12 parts of Terminalia chebula, 5 parts of Cuttlebone, and 12 parts of Radix Glycyrrhizae Preparata.

[0047] Example 7

[0048] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 25 parts of processed aconite, 22 parts of epimedium, 17 parts of Cistanche deserticola, 26 parts of Magnolia officinalis, 24 parts of Cyperus rotundus, 24 parts of Acanthopanax senticosus, 25 parts of Zedoariae, 25 parts of Herba Euphorbiae, 28 parts of Terminalia chebula, 16 parts of Cuttlebone, and 26 parts of Radix Glycyrrhizae Preparata.

[0049] Example 8

[0050] The difference between this embodiment and embodiment 1 is that the Chinese medicine preparation is composed of the following raw materials in weight fractions: 23 parts of processed aconite, 20 parts of epimedium, 16 parts of Cistanche deserticola, 24 parts of Magnolia officinalis, 22 parts of Cyperus rotundus, 18 parts of Acanthopanax senticosus, 20 parts of Zedoariae, 24 parts of Herba Euphorbiae, 24 parts of Terminalia chebula, 14 parts of Cuttlebone, and 24 parts of Radix Glycyrrhizae Preparata.

[0051] Test Example 1

[0052] 1 Materials and methods

[0053] 1.1 Cells

[0054] Rat IEC-6 cells were purchased from Shanghai Cell Bank.

[0055] 1.2 Preparation of Chinese medicine stock solution of the present invention

[0056] Experimental group 1: 20 parts of processed aconite root, 15 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of ginger and magnolia bark, 15 parts of Cyperus rotundus, 15 parts of Acanthopanax senticosus, 15 parts of Eupatorium, 15 parts of Herba Euphorbiae, 20 parts of Terminalia chebula, 10 parts of Cuttlebone, and 20 parts of roasted Licorice root.

[0057] Experimental group 2: 10 parts of Cistanche deserticola, 20 parts of Magnolia officinalis, 15 parts of Cyperus rotundus, 15 parts of Acanthopanax senticosus, 15 parts of Eupatorium fortunei, 15 parts of Herba Euphorbiae Ichthyophthirius, 20 parts of Terminalia chebula, 10 parts of Cuttlebone, and 20 parts of Radix Glycyrrhizae Preparata.

[0058] Experimental group 3: 20 parts of processed aconite root, 15 parts of epimedium, 15 parts of Acanthopanax senticosus, 15 parts of Zedoariae, 15 parts of Herba Euphorbiae, 20 parts of Terminalia chebula, 10 parts of Cuttlebone, and 20 parts of roasted Licorice root.

[0059] Experimental group 4: 20 parts of processed aconite root, 15 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of ginger and magnolia bark, 15 parts of Cyperus rotundus, 20 parts of Terminalia chebula, 10 parts of Cuttlebone, and 20 parts of roasted Licorice root.

[0060] Experimental group 5: 20 parts of processed aconite root, 15 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of ginger magnolia bark, 15 parts of Cyperus rotundus, 15 parts of Acanthopanax senticosus, 15 parts of Eupatorium, and 15 parts of Euphorbia pulegosa.

[0061] Weigh each raw material according to the above-mentioned weight proportions, wash, dry, and grind into powder, add 10 times (m / V) amount of distilled water, soak for 30 minutes, decoct for 1 hour, obtain filtrate, add 8 times amount of distilled water again and decoct for 1 hour, combine the filtrate, and concentrate to a drug mass concentration of 1 g / mL to obtain the test 1-5 groups of Chinese medicine stock solution, and place in a 4°C refrigerator for use.

[0062] 1.3 Cell culture

[0063] IEC-6 cells were inoculated in DMEM high-glucose medium containing 10% FBS and 1% double antibody (hereinafter referred to as "complete medium"), cultured at 37°C and 5% CO2 saturated humidity, and the medium was changed every 2-3 days. Subsequent experiments were performed after the cells grew to the logarithmic phase.

[0064] 1.4 Effects of each group of drugs on H2O2-induced oxidative stress in IEC-6 cells

[0065] IEC-6 cells in the logarithmic growth phase were taken and 1×10 5 / mL were inoculated in a 96-well plate, 100μL per well. After the cells adhered to the wall, they were divided into a blank control group, a model control group, and test groups 1-5, with 3 replicates in each group. 200μL of complete medium was added to the blank control group, 200μL of complete medium containing 60μmol / L H2O2 was added to the model control group, and 200μL of complete medium containing 60μmol / L H2O2 and 50μmol / L of the original Chinese medicine solution of test groups 1-5 were added to the test groups 1-5, respectively. After continuing to culture for 24h, the cell supernatant was collected, and the MDA content and SOD activity were detected using an ELISA reader according to the instructions of the kit.

[0066] 1.5 Effects of each group of drugs on LPS-induced IEC-6 cell secretion of inflammatory cytokines

[0067] IEC-6 cells in the logarithmic growth phase were taken and 1×10 5 / mL were inoculated in a 96-well plate, 100μL per well. After the cells adhered to the wall, they were divided into a blank control group, a model control group, and test groups 1-5, with 3 replicates in each group. 50μL of complete medium containing 50μmol / L of the original solution of the Chinese medicine in test groups 1-5 was added to the test groups 1-5, and the blank control group and the model control group were given an equal volume of complete medium. After 1h, 50μL of LPS with a final concentration of 1μg / mL was added to each well for stimulation, and the supernatant was collected after continued culture for 24h, and the secretion levels of TNF-α and IL-1β in the supernatant were determined according to the instructions of the ELISA kit.

[0068] 1.6 Statistical methods

[0069] All data were processed using Graphpad Prism 8.0 software and presented as mean ± standard deviation. When the data series of each group were normally distributed, ANOVA was used for variance analysis. When the variances were equal, LSD-test was used for inter-group comparison; when the variances were unequal, Dunnett's test was used for inter-group comparison. The difference was considered statistically significant when P < 0.05.

[0070] 2 Results

[0071] 2.1 Effects of each group of drugs on H2O2-induced oxidative stress in IEC-6 cells

[0072] The effects of each group of drugs on the oxidative stress of IEC-6 cells induced by H2O2 are shown in Table 1. Compared with the blank control group, the MDA content in the model control group was significantly increased, and the SOD activity was significantly decreased (P<0.01); compared with the model control group, the MDA content in the IEC-6 cells of the test groups 1-5 was significantly decreased (P<0.01), and the SOD activity increased to varying degrees. Among them, the test group 1 showed the best antioxidant effect, with the largest reduction in MDA content and the highest increase in SOD activity. Due to the lack of some Chinese medicine components, the antioxidant effects of the test groups 2-5 were inhibited to varying degrees, indicating that the Chinese medicine preparation of the present invention can inhibit the oxidative stress damage of IEC-6 cells induced by H2O2, and has a potential antioxidant stress effect. The Chinese medicine preparation of the present invention is treated according to the syndrome differentiation of traditional Chinese medicine and is reasonably formulated, and each component is indispensable.

[0073] Table 1 Comparison of MDA content and SOD activity in IEC-6 cells of each group

[0074] Grouping MDA content (nmol / mL) SOD activity (U / mL) Blank control group 4.15±0.11 0.49±0.07 Model control group <![CDATA[9.24±0.10 ## ]]> <![CDATA[0.13±0.04 ## ]]> Test Group 1 <![CDATA[5.33±0.25 ** ]]> <![CDATA[0.41±0.04 ** ]]> Test Group 2 <![CDATA[7.54±0.13 ** ]]> 0.19±0.01 Experiment 3 groups <![CDATA[7.05±0.07 ** ]]> <![CDATA[0.26±0.02 ** ]]> Test 4 groups <![CDATA[6.51±0.12 ** ]]> <![CDATA[0.30±0.02 ** ]]> Test 5 groups <![CDATA[6.31±0.18 ** ]]> <![CDATA[0.34±0.03 ** ]]>

[0075] Note: Compared with the blank control group, # P<0.05, ## P<0.01; compared with the model control group, * P<0.05,** P<0.01.

[0076] 2.2 Effects of each group of drugs on the secretion of inflammatory factors by IEC-6 cells induced by LPS

[0077] The effects of each group of drugs on the secretion of inflammatory factors by IEC-6 cells induced by LPS are shown in Table 2. Compared with the blank control group, the TNF-α and IL-1β contents in the model control group were significantly increased (P<0.01); compared with the model control group, the TNF-α and IL-1β in the test groups 1-5 were significantly decreased (P<0.05 or P<0.01), among which the test group 1 showed the best anti-inflammatory effect, and its TNF-α and IL-1β secretion levels decreased the most. Due to the lack of some Chinese medicine components, the inhibitory effect of the test groups 2-5 on the secretion level of inflammatory factors was weakened, indicating that the Chinese medicine preparation of the present invention can effectively inhibit the secretion of inflammatory factors by IEC-6 cells induced by LPS, thereby alleviating the inflammatory response, and has a potential effect in treating ulcerative colitis. The Chinese medicine preparation of the present invention is based on the unique syndrome differentiation and treatment system of traditional Chinese medicine, and is carefully formulated, and each component is indispensable.

[0078] Table 2 Comparison of the levels of inflammatory factors secreted by IEC-6 cells in each group

[0079]

[0080]

[0081] Note: Compared with the blank control group, # P<0.05, ## P<0.01; compared with the model control group, * P<0.05, ** P<0.01.

[0082] 3 Conclusion

[0083] The Chinese medicine preparation of the present invention can significantly reduce the MDA content in IEC-6 cells induced by H2O2, enhance the SOD activity, and effectively inhibit the oxidative stress damage of IEC-6 cells induced by H2O2; at the same time, the Chinese medicine preparation of the present invention can also significantly reduce the levels of inflammatory factors TNF-α and IL-1β secreted by IEC-6 cells induced by LPS, thereby alleviating the inflammatory response. These results show that the Chinese medicine preparation of the present invention has potential application value in the treatment of ulcerative colitis.

[0084] Test Example 2

[0085] 1 Materials and methods

[0086] 1.1 Experimental animals

[0087] 72 SPF male C57BL / 6N mice, 8 weeks old, weighing 22-24g. The experimental mice were raised in an SPF animal room with a 12-hour day and night cycle, a temperature of 20-26℃, and a humidity of 40-70%. The experiment was officially carried out after one week of adaptive feeding.

[0088] 1.2 Experimental Reagents

[0089] Dextran sulfate sodium (DSS), purchased from MP Biomedicals, batch number 0216011080;

[0090] Isoflurane for pets was purchased from Shenzhen Reward Life Science Co., Ltd., batch number R510-22-10;

[0091] The modified Sirius red staining kit was purchased from Beijing Solebow Technology Co., Ltd., batch number G1472;

[0092] PBS buffer, purchased from Gibco, batch number 20012027;

[0093] Glycogen PAS staining kit was purchased from Beijing Solebow Technology Co., Ltd., batch number G1281.

[0094] 1.3 Experimental drugs

[0095] 20 parts of processed aconite, 15 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of ginger magnolia bark, 15 parts of cyperus, 15 parts of Acanthopanax senticosus, 15 parts of zedoaria, 15 parts of euphorbia, 20 parts of terminalia chebula, 10 parts of sea cucumber, and 20 parts of licorice. According to the above weight fraction, each raw material is weighed, washed, dried, and powdered, and 10 times (m / V) of distilled water is added, soaked for 30 minutes, decocted for 1 hour, and a filtrate is obtained. 8 times of distilled water is added again and decocted for 1 hour, and the filtrate is combined and concentrated to a drug mass concentration of 1g / mL to obtain the Chinese medicine stock solution of the present invention, which is stored in a 4°C refrigerator for standby use.

[0096] Mesalazine enteric-coated tablets were purchased from Jiamusi Luling Pharmaceutical Co., Ltd. of Sunflower Pharmaceutical Group.

[0097] 1.4 Animal modeling, grouping and drug administration

[0098] After one week of adaptive feeding, mice were randomly divided into a control group, a model group, a mesalazine group, a low-dose (TL) group of the Chinese medicine preparation of the present invention, a medium-dose (TM) group of the Chinese medicine preparation of the present invention, and a high-dose (TH) group of the Chinese medicine preparation of the present invention according to body weight, with 12 mice in each group. The drinking water of mice in the remaining groups except the control group was replaced with a 3% DSS solution. The administration began on the day of modeling, and the mesalazine group was gavaged with 678 mg / kg / d mesalazine enteric-coated tablets, the low-dose group of the Chinese medicine preparation of the present invention, the medium-dose group of the Chinese medicine preparation of the present invention, and the high-dose group of the Chinese medicine preparation of the present invention were gavaged with 1000 mg / kg / d, 2000 mg / kg / d, and 4000 mg / kg / d of the Chinese medicine stock solution of the present invention, respectively, and the control group and the model group were gavaged with equal volumes of ultrapure water, once a day, for 7 consecutive days.

[0099] 1.5 Detection indicators

[0100] 1.5.1 Disease Activity Index

[0101] From the first day of modeling, the body weight, stool characteristics and bloody stool of mice were recorded and scored every day. According to the disease activity index (DAI) scoring standard, two experimenters performed double-blind scoring on the modeling mice, and finally took the average of the percentage of body weight loss, stool characteristics and bloody stool scores as the DAI score, and the scoring results were statistically analyzed.

[0102] Table 1 DAI scoring criteria

[0103] Weight loss Stool form Blood in the stool 0 0-1％ Normal morphology No blood in stool 1 1％-5％ Slightly loose stools Mild blood in stool 2 5％-10％ Small amount of loose stools Small amount of blood in stool 3 10％-18％ Lots of loose stools Heavy blood in the stool 4 Greater than 18% Severe diarrhea Blood fills the entire colon

[0104] 1.5.2 Colon gross observation

[0105] Clinically, the continuous chronic inflammatory response of the colonic mucosa of UC patients leads to fibrosis of the colonic mucosal interstitium, which in turn affects the function of the colonic smooth muscle. The colonic muscle layer gradually loses its motility, which eventually leads to shortening of the colon in UC patients. Therefore, the colon morphology of each group of mice was observed and the length was measured during the autopsy. After the last administration, the mice were exposed to excessive CO2 and killed. After the abdomen was opened, all the intestinal segments from the lower end of the cecum to the pubic symphysis were removed, and the fat outside the intestinal wall was carefully removed. The tissue was rinsed in pre-prepared saline, and the water was absorbed with absorbent paper. The colon of each group of mice was photographed, and the natural length of the colon of each mouse was recorded.

[0106] 1.5.3 Detection of inflammatory factors in plasma and colon tissue

[0107] After the last administration, blood was collected from the abdominal aorta of the mouse and placed in an EDTA anticoagulant tube. The blood was gently shaken up and down 2-3 times and mixed for 30 minutes. The blood was centrifuged at 4°C and 3000 r / min for 10 minutes. The supernatant was taken to obtain the plasma sample and stored in a -80°C refrigerator.

[0108] Accurately weigh the mouse colon tissue, add PBS at a ratio of weight (g): volume (mL) = 1:18, put in grinding beads, grind in a freezing grinder, centrifuge at 3500r / min at 4°C for 20min, and dispense the supernatant into EP tubes to obtain colon tissue homogenate.

[0109] The contents of IL-6, IL-10, IL-1β and TNF-α in plasma and colon tissue homogenate were detected according to the instructions of the ELISA kit.

[0110] 1.5.4 Pathological examination of colon tissue HE staining

[0111] The colon tissue of about 0.5 cm from the end was fixed in 4% paraformaldehyde for 48 h, and then cut into blocks and dehydrated in different concentrations of ethanol. The dehydrated tissue was embedded in paraffin and sliced ​​at a thickness of 4 μm. The pathological morphological changes of the colon tissue were observed under a microscope after HE staining, and the colon pathological injury score (Colonic mucosal damage index, CMDI) was calculated for the colon pathological sections according to Table 2.

[0112] Table 2 CMDI scoring criteria

[0113]

[0114] 1.6 Data Processing and Statistics

[0115] All data were processed using Graphpad Prism 8.0 software and presented as mean ± standard deviation. When the data series of each group were normally distributed, ANOVA was used for variance analysis. When the variances were equal, LSD-test was used for inter-group comparison. When the variances were unequal, Dunnett's was used for inter-group comparison. When one or more data series were non-normally distributed, non-parametric tests were used. Non-parametric tests (Kruskal-Wallis H rank sum test) were used for ranked data. Data were expressed as Md(P 25 ,P 75 )express.

[0116] 2 Results

[0117] 2.1 Effects on clinical symptoms and DAI scores of UC mice

[0118] The weight of mice in the Control group increased steadily, their fur was smooth and shiny, they had no abnormal behavior, and their feces were granular, dry, and without blood in the stool. Compared with the Control group, the clinical symptoms of ulcerative colitis in the mice in the modeling group became increasingly severe. The mice had dull fur, slow movements, listlessness, curled-up bodies, and a significant decrease in weight. Their feces were loose or watery, and they even had blood in the stool.

[0119] As shown in Table 3, compared with the Control group, the DAI score of the mice in the Model group was significantly increased (P<0.01); compared with the Model group, the DAI score of the mice in each drug-treated group was significantly decreased (P<0.01 or P<0.05), among which, the TH dose group had the most obvious improvement on the clinical symptoms of the ulcerative colitis model mice, and the decrease in the DAI score was also the most significant, indicating that the Chinese medicine preparation of the present invention has a significant improvement effect on the clinical symptoms of the ulcerative colitis model mice.

[0120] Table 3 Comparison of DAI scores of mice in each group [Md(P 25 ,P 75 )]

[0121]

[0122] Note: Compared with the Control group, ## P<0.01, # P<0.05; compared with the Model group, ** P<0.01, * P<0.05.

[0123] 2.2 Effects on colon length in UC mice

[0124] Depend on Figure 1 As shown in Table 4, compared with the Control group, the colon length of the mice in the Model group was significantly shortened (P<0.01), and obvious edema and bleeding occurred; compared with the Model group, the colon length of the mice in each drug-treated group was significantly increased (P<0.01), among which the TH dose group showed a more significant effect in the recovery of colon length, which was close to the colon length of the Control group. In addition, from the appearance of the colon, the symptoms of colon edema and bleeding in the mice in each drug-treated group were significantly alleviated, indicating that the Chinese medicine preparation of the present invention can significantly improve colon tissue damage and has a significant therapeutic effect in the treatment of ulcerative colitis.

[0125] Table 4 Comparison of colon length of mice in each group ( cm)

[0126] Grouping n Dosage (mg / kg) Colon length Control 12 - 6.55±0.39 Model 12 - <![CDATA[4.55±0.39 ## ]]> Etiasa 12 678 <![CDATA[5.12±0.46 ** ]]> TL 12 1000 <![CDATA[5.12±0.52 ** ]]> TM 12 2000 <![CDATA[5.19±0.34 ** <!-- 9 -->]]> TH 12 4000 <![CDATA[5.42±0.29 ** ]]>

[0127] Note: Compared with the Control group, ##P<0.01, # P<0.05; compared with the Model group, ** P<0.01, * P<0.05.

[0128] 2.3 Effects on the expression of inflammatory cytokines in plasma and colon tissue of UC mice

[0129] As shown in Table 5, compared with the Control group, the expression levels of pro-inflammatory cytokines IL-6, TNF-α and IL-1β in the plasma of the Model group mice were significantly increased, and the level of the anti-inflammatory cytokine IL-10 was significantly decreased (P<0.05 or P<0.01); compared with the Model group, the levels of IL-6, TNF-α and IL-1β in the Etiasa group and the TH dose group mice were significantly decreased, and the level of IL-10 was significantly increased (P<0.05 or P<0.01); the levels of IL-1β and TNF-α in the TM group mice were significantly decreased (P<0.05 or P<0.01); the level of IL-1β in the TL group mice was significantly decreased (P<0.05); the TM group and the TL group mice had a tendency to reduce the expression level of IL-6 and increase the expression level of IL-10 (P>0.05); the TL group mice had a tendency to reduce the expression level of TNF-α (P>0.05). The above results show that the Chinese medicine preparation of the present invention can regulate the inflammatory cytokines in the plasma of ulcerative colitis model mice and alleviate the inflammatory response.

[0130] As shown in Table 6, compared with the Control group, the content of proinflammatory cytokines IL-6, TNF-α and IL-1β in the colon tissue of the Model group mice was significantly increased, and the content of IL-10 was significantly decreased (P<0.05 or P<0.01); compared with the Model group, the content of IL-6, TNF-α and IL-1β in the Etiasa group and TH group mice was significantly decreased, and IL-10 was significantly increased (P<0.05 or P<0.01); the content of IL-6 and TNF-α in the TM group and TL group mice was significantly decreased, and IL-10 was significantly increased (P<0.05 or P<0.01); the TM group and TL group had a tendency to reduce the expression level of IL-1β in the colon tissue of mice (P>0.05). The above results show that the Chinese medicine preparation of the present invention can reduce the inflammatory response of the colon tissue of the ulcerative colitis model mice.

[0131] Table 5 Comparison of plasma IL-6, TNF-α, IL-1β and IL-10 levels in mice of each group ( n=12)

[0132] Group IL-6 (pg / mL) TNF-α(pg / mL) IL-1β (pg / mL) IL-10 (pg / mL) Control 25.64±11.47 45.48±9.27 8.60±3.96 658.73±243.65 Model <![CDATA[55.85±28.75 # ]]> <![CDATA[99.25±27.56 ## ]]> <![CDATA[23.73±12.48 ## ]]> <![CDATA[354.65±76.15 ## ]]> Etiasa <![CDATA[28.65±5.07 * ]]> <![CDATA[57.32±21.96 ** ]]> <![CDATA[10.75±6.54 * ]]> <![CDATA[515.75±119.73 ** ]]> TL 53.90±16.66 83.55±4.32 <![CDATA[13.85±4.75 * ]]> 438.85±212.25 TM 47.75±19.12 <![CDATA[62.02±9.75 ** ]]> <![CDATA[11.85±6.88 * ]]> 399.22±102.78 TH <![CDATA[29.75±14.34 * ]]> <![CDATA[60.15±6.32 ** ]]> <![CDATA[11.74±6.20 * ]]> <![CDATA[520.14±154.10 * ]]>

[0133] Note: Compared with the Control group, ## P<0.01, #P<0.05; compared with the Model group, ** P<0.01, * P<0.05.

[0134] Table 6 Comparison of IL-6, TNF-α, IL-1β and IL-10 contents in colon tissue homogenates of mice in each group ( n=12)

[0135] Group IL-6 (pg / mg) TNF-α(pg / mg) IL-1β (pg / mg) IL-10 (pg / mg) Control 6.71±3.40 3.76±2.97 3.14±2.83 86.52±33.12 Model <![CDATA[13.69±3.10 ## ]]> <![CDATA[10.89±4.56 ## ]]> <![CDATA[15.76±10.93 # ]]> <![CDATA[32.17±11.22 ## ]]> Etiasa <![CDATA[7.87±2.18 ** ]]> <![CDATA[4.63±3.34 ** ]]> <![CDATA[3.40±1.81 * ]]> <![CDATA[59.54±26.33 * ]]> TL <![CDATA[8.78±3.45 * ]]> <![CDATA[6.56±2.32 * ]]> 11.20±3.19 <![CDATA[60.53±15.65 ** ]]> TM <![CDATA[9.46±3.15 * ]]> <![CDATA[5.69±3.00 * ]]> 5.63±2.42 <![CDATA[52.30±11.15 * ]]> TH <![CDATA[9.25±2.20 ** ]]> <![CDATA[5.25±1.19 ** ]]> <![CDATA[4.75±1.70 * ]]> <![CDATA[52.81±4.51 * ]]>

[0136] Note: Compared with the Control group, ## P<0.01, # P<0.05; compared with the Model group, ** P<0.01, * P<0.05.

[0137] 2.4 Effect of the Chinese medicine preparation of the present invention on pathological damage of colon tissue in ulcerative colitis mouse model

[0138] Histopathological examination results Figure 2 As shown. The mucosal layer, submucosal layer, muscular layer and outer membrane of the colon of the mice in the Control group had clear morphology, complete structure, regular cell arrangement, complete intestinal structure, no edema and thickening of the submucosal layer, no lymphocyte aggregation and inflammatory cell infiltration at the intestinal base, and neatly arranged intestinal crypts. The local mucosal epithelium of the mice in the Model group fell off, the epithelium was damaged, a large number of inflammatory cells infiltrated into the submucosal layer, the intestinal glands were significantly expanded, the number of intestinal crypts was significantly reduced, the intestinal submucosal layer was edematous and thickened, and the crypts were twisted and atrophied. The mucosal layer, submucosal layer, muscular layer and outer membrane of the colon of the mice in the Etiasa group had clear morphology, complete structure, regular cell arrangement, no obvious cell degeneration, and no obvious inflammatory cell infiltration. The colon mucosal epithelium of the mice in the TH group did not fall off, the mucosal epithelium structure was relatively complete, the intestinal mucosa was not significantly thickened, the cells were relatively regularly arranged, the intestinal glands were not significantly expanded, and a small number of inflammatory cells infiltrated the submucosal layer. The colon mucosal epithelium of mice in the TM group did not fall off, the mucosal epithelium structure was relatively intact, the intestinal mucosa did not show obvious thickening, the intestinal glands did not show obvious expansion, and inflammatory cells infiltrated into the submucosal layer. The colon mucosal epithelium of mice in the TL group did not fall off, the mucosal stroma thickened, inflammatory cells infiltrated into the submucosal layer, and the number of crypts decreased.

[0139] As shown in Table 7, compared with the Control group, the CMDI score of the mice in the Model group was significantly increased, and the difference was statistically significant (P<0.01); compared with the Model group, the CMDI scores of the mice in the Etiasa, TM and TH dose groups were significantly decreased, and the difference was statistically significant (P<0.01); the CMDI score of the mice in the TL dose group tended to decrease (P>0.05). The above results indicate that the Chinese medicine preparation of the present invention can repair the colon tissue damage caused by DSS.

[0140] Table 7 Effect of CMDI scores of colon pathology in each group of mice [Md (P 25 ,P 75 )]

[0141] Group n Dosage (mg / kg) CMDI score Control 12 - 0.00(0.00,0.00) Model 12 - <![CDATA[2.67(2.33,3.00) ## ]]> Etiasa 12 678 <![CDATA[1.33(0.67,1.33) ** ]]> TL 12 1000 2.33(2.00,2.33) TM 12 2000 <![CDATA[1.33(1.00,1.67) ** ]]> TH 12 4000 <![CDATA[1.00(1.00,1.33) ** ]]>

[0142] Note: Compared with the Control group, ## P<0.01, # P<0.05; compared with the Model group, ** P<0.01, * P<0.05.

[0143] 3 Conclusion

[0144] In summary, the Chinese medicine preparation of the present invention has a significant therapeutic effect on ulcerative colitis model mice, can significantly reduce the DAI scores of model mice, can repair and improve the integrity of colon tissue structure, enhance intestinal barrier function, can inhibit the expression of proinflammatory cytokines, effectively improve inflammatory response, has a significant antioxidant effect, and provides a new treatment option for the treatment of ulcerative colitis.

[0145] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A Chinese medicine preparation for treating ulcerative colitis, characterized in that: The traditional Chinese medicine preparation is composed of the following raw materials in weight fractions: 10-28 parts of processed aconite root, 8-26 parts of epimedium, 3-22 parts of cistanche deserticola, 12-30 parts of magnolia officinalis, 5-28 parts of cyperus rotundus, 5-27 parts of acanthopanax senticosus, 5-27 parts of zedoaria, 8-27 parts of euphorbia cerevisiae, 10-32 parts of terminalia chebula, 3-20 parts of cuttlebone and 10-32 parts of roasted liquorice.

2. A Chinese medicine preparation for treating ulcerative colitis according to claim 1, characterized in that: The traditional Chinese medicine preparation is composed of the following raw materials in weight fractions: 15-25 parts of processed aconite root, 10-20 parts of epimedium, 5-15 parts of cistanche deserticola, 15-25 parts of zingiber officinale, 10-20 parts of cyperus rotundus, 10-20 parts of acanthopanax senticosus, 10-20 parts of zedoaria, 10-20 parts of euphorbia pulex, 10-20 parts of euphorbia chebula, 15-25 parts of terminalia chebula, 5-15 parts of cuttlebone and 15-25 parts of roasted liquorice.

3. A Chinese medicine preparation for treating ulcerative colitis according to claim 1, characterized in that: The Chinese medicine preparation is composed of the following raw materials in weight fractions: 20 parts of processed aconite root, 15 parts of epimedium, 10 parts of Cistanche deserticola, 20 parts of ginger magnolia bark, 15 parts of cyperus rotundus, 15 parts of acanthopanax senticosus, 15 parts of zedoaria, 15 parts of euphorbia pulex, 20 parts of terminalia chebula, 10 parts of cuttlebone and 20 parts of roasted liquorice.

4. A Chinese medicine preparation for treating ulcerative colitis according to any one of claims 1 to 3, characterized in that: The Chinese medicine preparation is prepared into any oral dosage form acceptable to the human body, including pills, powders, granules, oral liquids, decoctions, and tablets, by conventional pharmaceutical methods and with medically acceptable adjuvants.

5. Use of the Chinese medicine preparation according to any one of claims 1 to 3 in the preparation of a medicament for preventing and / or treating ulcerative colitis.

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