Application of elm and gardenia hemostasis granules in treatment of enteritis

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a novel medical application of elm and gardenia hemostasis granules, in particular to an application of the elm and gardenia hemostasis granules in medicines for treating inflammatory intestinal diseases. Pharmacological experiments show that the ulcerative colitis can be effectively improved, inflammatory factors TNF-alpha and IL-1beta in serum of rats can be reduced, the ulcerative colitis can be immunologically regulated, MPO and SOD activities of colon tissue of mice can be regulated, the oxidative stress resistance of colon can be improved, oxidative damage of colon can be relieved, inflammation can be relieved, and the ulcerative colitis can be effectively treated. The traditional Chinese medicine composition is convenient to take, high in safety, small in adverse reaction, wide in application prospect and worthy of clinical popularization and application.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical technology, and particularly relates to a new pharmaceutical use of Yuzhi Zhixue Granules, and provides the use of Yuzhi Zhixue Granules in the treatment of inflammatory bowel diseases. Background Art

[0002] Inflammatory bowel disease (IBD) is a chronic and recurrent gastrointestinal inflammatory disease, and ulcerative colitis (UC) and Crohn's disease (CD) are the most common. Among them, ulcerative colitis (UC) is an intestinal disease caused by chronic non-specific inflammatory reactions in the colorectum, mainly occurring in the colonic mucosa layer, mainly with ulcer erosion. It can involve the sigmoid colon and even spread to the entire colon, and can develop into colon cancer in severe cases. In recent years, the prevalence of UC at home and abroad has shown a gradually increasing trend and is considered a global disease. Clinically, UC patients often show bloody diarrhea, tenesmus, increased frequency of bowel movements, weight loss, etc. Its etiology is not yet clear, the pathogenesis is complex, and the condition is lingering and difficult to cure.

[0003] Currently, Western medicine treatment mainly uses drugs such as hormones, sulfonamides, and immunosuppressants. However, hormones and sulfonamides have relatively large adverse reactions, and there are dependence and stimulant resistance, making it difficult to use them for a long time. Moreover, immunosuppressants are expensive and their targeting needs to be improved. Therefore, there is currently no clear treatment method in Western medicine, while traditional Chinese medicine believes that liver depression and spleen deficiency can cause ulcerative colitis. At present, there are insufficient treatment means to cure it, so inducing and maintaining symptom remission and preventing disease recurrence have become the goals of drugs at the present stage.

[0004] In recent years, with the continuous in-depth research on therapeutic drugs, the advantages of traditional Chinese medicine with small side effects and good curative effects have gradually emerged. Therefore, it is of great significance to study more traditional Chinese medicine compositions that can be used to treat ulcerative colitis. The drug composition of Yuzhi Zhixue Granules includes Sanguisorba officinalis, Eclipta prostrata, Gardenia jasminoides, Agrimonia pilosa, Sophora japonica, Bistorta officinalis, Rubia cordifolia, Typha angustifolia, Rehmannia glutinosa, Paeonia lactiflora, Scutellaria baicalensis, Angelica sinensis, Paeonia suffruticosa, Dryopteris crassirhizoma, Cirsium japonicum, and Platycladus orientalis. The various herbs in the formula are combined to have the effect of clearing heat and stopping bleeding. However, the effect of Yuzhi Zhixue Granules on inflammatory bowel diseases has not been reported yet. Exploring the treatment of inflammatory bowel diseases by traditional Chinese medicine undoubtedly has greater significance, and the research in this area will surely have a broader prospect. Summary of the Invention

[0005] The present invention is a further development of the use of the existing product "Yuzhi Zhixue Granules". It is found that Yuzhi Zhixue Granules have the effects of clearing heat and stopping bleeding and reducing cellular inflammatory factors. Based on this effect, the inventor has further studied and developed Yuzhi Zhixue Granules and its formula for the treatment of inflammatory bowel diseases.

[0006] The object of the present invention is to provide a new use of Yuzhi Zhixue Granules.

[0007] The present invention provides the use of Yuzhi Zhixue Granules in the preparation of a medicament for treating inflammatory bowel disease.

[0008] According to some embodiments, the above-mentioned inflammatory bowel disease includes ulcerative colitis, Crohn's disease and indeterminate colitis.

[0009] According to some embodiments, the ulcerative colitis described in the present invention is differentiated in traditional Chinese medicine as one or more of internal accumulation of dampness and heat, qi stagnation and blood stasis, and spleen-kidney yang deficiency.

[0010] According to some embodiments, the ulcerative colitis described in the present invention is manifested as one or more of bloody diarrhea, abdominal pain, hematochezia, tenesmus, and vomiting.

[0011] According to some embodiments, the ulcerative colitis described in the present invention is clinically divided into initial onset type, chronic recurrence type, chronic persistent type, and acute type.

[0012] According to some embodiments, the medicament described in the present invention has an immunomodulatory effect and can reduce inflammatory factors and tumor necrosis factor in the serum.

[0013] According to some embodiments, the inflammatory factors described in the present invention include but are not limited to IL-1β, and the tumor necrosis factor is TNF-α.

[0014] According to some embodiments, the medicament described in the present invention can improve the oxidative stress ability of the colon.

[0015] According to some embodiments, the improvement of the oxidative stress ability of the colon described in the present invention is manifested as regulating the activities of MPO and SOD in the colon tissue.

[0016] The first discovery of the present invention: After treating ulcerative colitis model rats with Yuzhi Zhixue Granules, it can relieve the symptoms of colitis in rats, has an immunomodulatory effect on ulcerative colitis, can reduce the level of inflammatory factors in the body (P < 0.05), inhibit the increase in the levels of pro-inflammatory factors IL-1β and tumor necrosis factor TNF-α, and relieve inflammatory reactions and tissue damage. At the same time, it is found that after using Yuzhi Zhixue Granules, the activities of MPO and SOD in the body of mice can be increased (P < 0.05), indicating that Yuzhi Zhixue Granules can improve the antioxidant stress ability of the colon, reduce colon oxidative damage, and relieve inflammation. Therefore, Yuzhi Zhixue Granules have broad application prospects in the preparation of medicaments for treating inflammatory bowel disease.

[0017] Obviously, based on the above content of the present invention, according to the common general knowledge and conventional means in the art, without departing from the above basic technical idea of the present invention, various other forms of modifications, substitutions or changes can be made.

[0018] The following is a further detailed description of the above content of the present invention in the form of specific embodiments. However, it should not be understood that the scope of the above subject matter of the present invention is limited to the following embodiments. Technologies implemented based on the above content of the present invention all fall within the scope of the present invention. Description of the Drawings

[0019] Figure 1 Changes in the levels of TNF-α in the sera of rats in each group. Note: # Compared with the normal group, P < 0.05; * compared with the model group, P < 0.05.

[0020] Figure 2 Changes in the levels of IL-1β in the sera of rats in each group. Note: # Compared with the normal group, P < 0.05; * compared with the model group, P < 0.05.

[0021] Figure 3 Changes in the MPO activity in the intestinal tissues of rats in each group. Note: # Compared with the normal group, P < 0.05; * compared with the model group, P < 0.05.

[0022] Figure 4 Changes in the SOD activity in the intestinal tissues of rats in each group. Note: # Compared with the normal group, P < 0.05; * compared with the model group, P < 0.05. Specific Embodiments

[0023] The raw materials and equipment used in the present invention are all known products and are obtained by purchasing commercially available products.

[0024] Example 1: Animal Experiment on the Improvement of Enteritis by Yuzhi Zhixue Granules

[0025] 1. Experimental Materials

[0026] 1.1 Experimental animals: SPF-grade male SD rats, with a body weight of 200 ± 20 g, provided by Lunan Pharmaceutical Group Co., Ltd., animal license: SYXK (Lu) 2018 - 0008.

[0027] They were adaptively fed for one week at a temperature of 22 - 25°C, a relative humidity of 60% - 70%, a light-dark cycle of 12 h, and standard feeding for mice, with free access to food and water. The feeding and sacrifice of experimental animals were all carried out in accordance with the Guide for the Care and Use of Laboratory Animals.

[0028] 1.2 Experimental drugs and preparations: Positive: Mesalazine Enteric-coated Tablets (Heilongjiang Tianhong Pharmaceutical Co., Ltd.); DSS (Dextran Sulfate Sodium) was used to induce ulcerative colitis in mice. Yuzhi Zhixue Granules (Shandong Xinsdai Pharmaceutical Co., Ltd., batch number: 1472006006).

[0029] 2. Experimental Methods

[0030] 2.1 Grouping and model establishment:

[0031] Randomly select 10 rats as the blank group, and establish a ulcerative colitis model in the remaining 30 rats using DSS. Prepare 3.5% 7d and 5% 7d DSS aqueous solutions with distilled water. The blank group drinks normal water. Observe the body weight, stool characteristics, and blood in the stool of the rats daily. After 14 days of model establishment, evaluate the disease activity index (DAI) of all rats. Randomly select 1 rat from the model group and the blank group, take the colon tissue, observe the colon injury situation macroscopically, and determine that the model establishment is successful. After successful model establishment, randomly divide them into 3 groups: the model group, the mesalazine enteric-coated tablet group (hereinafter referred to as the positive group), and the Yuzhi Zhixue Granule group (hereinafter referred to as the Yuzhi group), with 10 rats in each group. Stop taking DSS, and each group resumes normal drinking water and is given the corresponding drugs for 14 days of treatment.

[0032] 2.2 Treatment plan:

[0033] In the positive group, after 24 hours of model establishment, administer mesalazine enteric-coated tablets by gavage. In the Yuzhi group, after 24 hours of model establishment, administer Yuzhi Zhixue Granules by gavage. After 24 hours of model establishment, the model group and the blank group are given an equal amount of normal saline by gavage. After the last gavage in each group of rats, fast the rats for 24 hours without restricting water intake, collect serum by orbital blood collection and sacrifice the mice, quickly store the colon tissue in liquid nitrogen, and then transfer it to a -80°C refrigerator for enzyme activity determination.

[0034] 2.3 Observation indicators

[0035] 2.3.1 Observe the DAI score of rats

[0036] Observe the general conditions of rats in each group, including mental state, hair color luster, activity, stool characteristics (characteristics, blood in the stool), and body weight. After successful model establishment, that is, after treatment, evaluate the disease activity index: DAI score. The calculation of the DAI score is shown in formula (1).

[0037]

[0038] Table 1 DAI scoring criteria

[0039] Score Rate of body weight change / % Stool character Hematochezia 0 ≥0 Normal feces None 1 0～-3 Soft feces Slight hematochezia 2 -3～-6 Semi-formed feces with mucus Patchy hematochezia 3 -6～-9 Loose feces Patchy hematochezia 4 -9～-12 Watery feces Hematochezia and anal swelling

[0040] 2.3.2 Determination of the levels of inflammatory factors in mouse serum

[0041] When the blood samples of each group of mice are left standing for 4 hours, after the blood coagulates, centrifuge at 3000 r / min for 15 minutes to take the upper-layer serum, dilute it to the required concentration with phosphate buffer (pH 7.2 - 7.4), and use the enzyme-linked immunosorbent assay method. Operate according to the kit instructions, measure OD450 nm with an enzyme-labeled instrument, and calculate the mass concentrations of TNF-α and IL-1β in each serum sample according to the standard curve.

[0042] 2.3.3 Determination of Oxidative Stress Level in Mouse Colon

[0043] Accurately weigh the colon tissue, add it to the homogenization medium in the detection kit and mix well. Operate according to the kit instructions. Use an enzyme-linked immunosorbent assay (ELISA) reader to measure the optical density (OD) at 460 nm, and determine and calculate the activities of myeloperoxidase (MPO) and superoxide dismutase (SOD) in the mouse colon tissue.

[0044] 2.4 Data Statistics and Analysis

[0045] The data are expressed as mean ± standard deviation. One-way analysis of variance in SPSS 21.0 was used for statistical analysis. A significant difference was defined as P < 0.05. Sigmaplot 14.0 was used for graphing.

[0046] 3. Results and Analysis

[0047] 3.1 Effects on DAI Score of Mice

[0048] Table 2 Effects of Yuzhi Zhixue Granules on DAI Score of Rats with Ulcerative Colitis (X±S)

[0049] Group Before treatment After treatment (14d) Blank group 0.00±0.00 0.00±0.00 Model group 3.65±0.20 2.88±0.31 Positive group 3.72±0.13 2.40±0.23 Yuzhi group 3.53±0.26 2.46±0.22

[0050] Before modeling, the rats in each group were in good health, with good mental state, sensitive reaction, smooth hair color, normal diet, normal stool in the shape of long strips, normal humidity, no bloody stool, normal weight and steady weight gain. Record the weight before treatment and conduct DAI scoring. After verifying the successful modeling, intragastric administration was carried out for 14 days. After the intervention treatment, DAI scoring was carried out again. In the model group, some rats still had loose stools and bloody stools. In the positive group and the Yuzhi group, the stools were dry and formed without bleeding. After treatment, the DAI scores of the positive group and the Yuzhi group were significantly different compared with those before treatment (P < 0.01). The results showed that Yuzhi Zhixue Granules might be able to relieve the symptoms of mouse colitis as well as mesalazine enteric-coated tablets.

[0051] 3.2 Immunomodulatory Effects on Cytokines in Ulcerative Colitis

[0052] The effects of Yuzhi Zhixue Granules on the levels of serum inflammatory factors TNF-α and IL-1β in rats are shown in Figure 1 、 Figure 2 .

[0053] Inflammatory factors are mainly secreted by immune system cells and can mediate the infiltration of inflammatory cells such as monocytes into the intestinal tissue, causing intestinal tissue damage and directly reflecting the degree of colon inflammation. Abnormal elevation of the secretion levels of pro-inflammatory factors such as TNF-α and IL-1β will exacerbate the inflammatory damage of the mouse colon.

[0054] Such as Figure 1 、 2As shown, compared with the normal group, the levels of pro-inflammatory factors in the serum of mice in the model group were significantly increased (P < 0.01). Compared with the model group, the levels of each inflammatory factor in the positive group and the Yuzhi group were significantly decreased (P < 0.01), and there was no significant difference between the positive group and the Yuzhi group (P > 0.05). Both could relieve the inflammatory response and tissue damage in the pathological process of DSS-induced ulcerative colitis in mice by inhibiting the increase in the levels of pro-inflammatory factors.

[0055] 3.3 Effects on oxidative stress in rat colon

[0056] The effects of Yuzhi Zhixue Granule on the activities of MPO and SOD in rat intestinal tissue are shown in Figure 3 、 4 。

[0057] The pathogenesis of ulcerative colitis is related to the imbalance of the intestinal antioxidant system. The accumulation of excessive reactive oxygen species and reactive nitrogen species will lead to the imbalance of the colon antioxidant defense system. As Figure 3 shown, compared with the normal group, the activity of MPO in the colon of mice in the model group was significantly increased (P < 0.01), while the activities of MPO in the positive group and the Yuzhi group were significantly lower than those in the model group (P < 0.01), and there was no significant difference between the positive group and the Yuzhi group (P > 0.05), indicating that the positive group and the Yuzhi group could effectively reduce neutrophils in the colon tissue of mice and reduce the degree of inflammation.

[0058] SOD can reduce oxidative stress damage by scavenging superoxide anion free radicals produced by the body. The decrease in SOD activity results in the decline of the colon's ability to scavenge free radicals and accelerates the damage of colon tissue. As Figure 4 shown, compared with the normal group, the SOD in the model group was significantly decreased (P < 0.01). Compared with the model group, the activities of SOD in the positive group and the Yuzhi group were significantly increased. Therefore, Yuzhi Zhixue Granule and the positive group can improve the antioxidant stress ability of the colon and relieve colon oxidative damage and inflammation by regulating the activity of SOD in the colon tissue of mice.

[0059] Based on the above experimental results, the present invention first discovers that Yuzhi Zhixue Granule can be used to prepare drugs for treating inflammatory bowel diseases, and has broad application prospects.

Claims

1. Use of a traditional Chinese medicine composition composed of Sanguisorba officinalis, Eclipta prostrata, Gardenia jasminoides, Cyrtomium fortunei, Agrimonia pilosa, Sophora japonica, Bistorta vivipara, Cirsium japonicum, Platycladus orientalis, Carbonized old palm, Cortex moutan, Rubia cordifolia, Typha angustifolia, Rehmannia glutinosa, Paeonia lactiflora, Scutellaria baicalensis, and Angelica sinensis in the preparation of a drug for treating inflammatory bowel disease.

2. The use according to claim 1, characterized in that, The described traditional Chinese medicine composition is prepared from the following raw materials: 5 - 200 parts by weight of Sanguisorba officinalis, 5 - 180 parts by weight of Eclipta prostrata, 5 - 260 parts by weight of Gardenia jasminoides 5 - 180 parts by weight of Sophora japonica, 5 - 100 parts by weight of Platycladus orientalis, 5 - 210 parts by weight of Typha angustifolia 5 - 220 parts by weight of Agrimonia pilosa, 5 - 120 parts by weight of Bistorta officinalis, 5 - 100 parts by weight of Cirsium japonicum 5 - 120 parts by weight of Carbonized Nuciferae Petiolus, 5 - 120 parts by weight of Cortex Moutan, 5 - 150 parts by weight of Rubia cordifolia 5 - 180 parts by weight of Rehmannia glutinosa, 5 - 100 parts by weight of Paeonia lactiflora, 5 - 160 parts by weight of Scutellaria baicalensis 5 - 160 parts by weight of Dryopteris crassirhizoma, 5 - 100 parts by weight of Angelica sinensis.

3. The use according to any one of claims 1 - 2, characterized in that, The described Sanguisorba officinalis is stir - fried Sanguisorba officinalis, the Gardenia jasminoides is stir - fried Gardenia jasminoides, the Sophora japonica is stir - fried Sophora japonica, the Platycladus orientalis is stir - fried Platycladus orientalis, the Typha angustifolia is stir - fried Typha angustifolia, and the Rubia cordifolia is red Rubia cordifolia.

4. The use according to claim 1, characterized in that, The described inflammatory bowel disease includes ulcerative colitis, Crohn's disease and indeterminate colitis.

5. The use according to claim 4, characterized in that, The traditional Chinese medicine syndrome differentiation of the described ulcerative colitis is one or more of internal accumulation of heat - dampness, qi stagnation and blood stasis, spleen - kidney yang deficiency.

6. The use according to claim 4, characterized in that, The described ulcerative colitis is clinically divided into initial type, chronic relapsing type, chronic persistent type, acute type.

7. The use according to any one of claims 1 - 6, characterized in that, The described drug has an immunomodulatory effect and can reduce inflammatory factors and tumor necrosis factor in the serum. Among them, the described inflammatory factors include but are not limited to IL - 1β, and the tumor necrosis factor is TNF - α.

8. The use according to any one of claims 1 - 6, characterized in that, The described drug can improve the oxidative stress ability of the colon.

9. The use according to claim 8, characterized in that, The improvement of the oxidative stress ability of the colon is manifested as regulating the activities of MPO and SOD in the colon tissue.

10. The use according to claim 1, characterized in that, The described traditional Chinese medicine composition is Yuzhi Zhixue Granules.

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