Application of avicularin in preparation of medicine for preventing and treating ulcerative colitis

Abstract

The invention relates to the technical field of ulcerative colitis prevention and treatment, in particular to application of avicularin in preparation of medicine for preventing and treating ulcerative colitis. The Avicularin can effectively improve mouse ulcerative colitis caused by dextran sodium sulfate (DSS), not only can improve colon injury and reduce a DAI index, but also can reduce expression of inflammatory cytokines and increase expression of intestinal mucosal barrier related protein.

Description

Technical Field

[0001] The invention relates to the technical field of prevention and treatment of ulcerative colitis, and in particular to the application of pygme pectin in the preparation of medicines for prevention and treatment of ulcerative colitis. Background Art

[0002] Ulcerative colitis (UC) is a refractory disease with abdominal pain, diarrhea, bloody stools and tenesmus as the main clinical manifestations, which seriously affects the patient's health and quality of life. Because it is difficult to cure and prone to recurrence and canceration, it has always been a hot topic in the field of digestive research. In recent years, with the improvement of living standards and changes in dietary structure and living habits, the incidence of ulcerative colitis in my country is increasing year by year.

[0003] The treatment goal of ulcerative colitis is to quickly induce remission and maintain remission, reduce hormone demand, resistance and dependence during treatment, prevent complications, and reduce the rate of surgery. Currently, commonly used Western medicine treatment methods include aminosalicylic acid preparations, immunosuppressants, hormones, biological preparations, etc., although certain therapeutic effects have been achieved, their remission rates are still unsatisfactory. In addition, these drugs may have different side effects, even serious adverse reactions. Traditional Chinese medicine has the advantages of high safety factor and few toxic and side effects, and the main effect is its effective ingredients. The application of reported effective ingredients of traditional Chinese medicine in the treatment of ulcerative colitis includes alkaloids, flavonoids, saponins, polysaccharides, such as berberine, baicalin, puerarin, ginsenosides, total glycosides of white paeony, astragalus polysaccharides, dandelion polysaccharides, etc.

[0004] Avicularin is mainly derived from the whole herb of Polygonum aviculare L., a plant of the Polygonaceae family, and is also found in many other plants. Avicularin is known to have a variety of biological activities, including anti-allergic, antioxidant, hepatoprotective, and anti-tumor effects. It has been reported that it can improve human hepatocellular carcinoma by regulating the activities of NF-κB (p65), cyclooxygenase-2 (COX-2), and peroxisome proliferator-activated receptor γ (PPAR-γ).

[0005] However, up to now, there is no report on the application of scutellaria baicalensis as an anti-enteritis drug. Summary of the invention

[0006] In view of this, the object of the present invention is to provide the use of scutellaria baicalensis in the preparation of drugs for preventing and treating ulcerative colitis.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0008] The invention provides application of scutellaria baicalensis in preparing medicine for preventing and treating ulcerative colitis.

[0009] Preferably, the ulcerative colitis is ulcerative colitis caused by dextran sulfate sodium.

[0010] Preferably, the glycoside exerts its effect by inhibiting the ERK / NF-κB signaling pathway.

[0011] Preferably, the dosage form of the drug for preventing and treating ulcerative colitis includes a suspension.

[0012] Preferably, when the dosage form of the drug for preventing and treating ulcerative colitis is a suspension, the preparation method of the drug for preventing and treating ulcerative colitis comprises the following steps:

[0013] After dissolving scutellaria baicalensis in sodium carboxymethylcellulose solution, the solution is diluted with water to obtain the drug for preventing and treating ulcerative colitis.

[0014] Preferably, the mass concentration of the sodium carboxymethyl cellulose solution is 0.4-0.6%.

[0015] The invention provides a medicine for preventing and treating ulcerative colitis, the effective ingredient of which is scutellaria baicalensis glycoside.

[0016] Preferably, auxiliary materials are also included.

[0017] Preferably, the dosage form of the drug comprises a suspension.

[0018] Preferably, when the dosage form of the drug is a suspension, the composition of the drug includes scutellaria baicalensis, sodium carboxymethyl cellulose and water.

[0019] The present invention provides the use of avicularin in the preparation of a drug for preventing and treating ulcerative colitis. Avicularin can effectively improve ulcerative colitis in mice caused by dextran sulfate sodium (DSS), not only improving colon damage and reducing the DAI index, but also reducing the expression of inflammatory cytokines and increasing the expression of intestinal mucosal barrier-related proteins. Avicularin exerts its anti-ulcerative colitis effect by inhibiting the ERK / NF-κB signaling pathway. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The following are some of the results of the prevention and treatment of DSS-induced ulcerative colitis by Avicularin, where A is the structural formula of Avicularin, B is the body weight test result, C is the blood stool result of mice, and D is the DAI index;

[0021] Figure 2 Another part of the results of Avicularin's prevention and treatment of DSS-induced ulcerative colitis, where A is the observation result of mouse colon, B is the result of mouse colon length, C is the result of colon histopathology score, and D is the result of HE staining of colon pathological section;

[0022] Figure 3 The intervention effect of Avicularin on the protein expression (A) and mRNA expression (B) of inflammatory cytokines IL-1β, IL-6, and TNF-α in mouse colon;

[0023] Figure 4 The results of Western Blot (WB) and immunofluorescence detection of tight junction proteins ZO-1 and occludin in colon tissue, where A is the protein expression level of tight junction proteins ZO-1 and occludin in colon tissue measured by WB, and B is the result of immunofluorescence detection of tight junction proteins ZO-1 and occludin in colon tissue;

[0024] Figure 5 These are the results of the effect of Avicularin on the MAPK / NF-κB signaling pathway, where A is the WB detection result of the MAPK signaling pathway, and B is the NF-κB signaling pathway detection result. DETAILED DESCRIPTION

[0025] The invention provides application of scutellaria baicalensis in preparing medicine for preventing and treating ulcerative colitis.

[0026] In the present invention, the molecular formula of the glycoside is C 20 H 18 O 11 , which can be obtained by purchase.

[0027] In the present invention, the ulcerative colitis is preferably ulcerative colitis caused by dextran sulfate sodium.

[0028] In the present invention, the scutellaria baicalensis exerts its effect by inhibiting the ERK / NF-κB signaling pathway.

[0029] Avicularin can effectively improve ulcerative colitis in mice induced by dextran sulfate sodium (DSS). It can not only improve colon damage and reduce the DAI index, but also reduce the expression of inflammatory cytokines and increase the expression of intestinal mucosal barrier-related proteins.

[0030] In the present invention, the dosage form of the drug for preventing and treating ulcerative colitis preferably includes a suspension; when the dosage form of the drug for preventing and treating ulcerative colitis is a suspension, the preparation method of the drug for preventing and treating ulcerative colitis preferably includes the following steps: dissolving pyrogallol in a sodium carboxymethyl cellulose solution, and then diluting with water to obtain the drug for preventing and treating ulcerative colitis.

[0031] In the present invention, the mass concentration of the sodium carboxymethyl cellulose solution is preferably 0.4-0.6%, more preferably 0.5%; the solvent of the sodium carboxymethyl cellulose solution is preferably water.

[0032] In the present invention, the water is preferably double distilled water (ddH2O).

[0033] The invention provides a medicine for preventing and treating ulcerative colitis, the effective ingredient of which is scutellaria baicalensis glycoside.

[0034] The present invention has no special requirements on the mass content of glycosides in the drug. In the present invention, based on the content of glycosides in the drug, the dosage of the drug is preferably 25 mg / kg to 50 mg / kg of mouse body weight.

[0035] In the present invention, the drug for preventing and treating ulcerative colitis preferably also includes excipients. The present invention has no special requirements on the types of the excipients, and any medical excipients known in the art can be used, such as excipients, antioxidants, and the like.

[0036] In the present invention, the dosage form of the drug preferably includes a suspension. When the drug is a suspension, the composition of the drug includes scutellaria baicalensis, sodium carboxymethylcellulose and water.

[0037] The application of the glycoside provided by the present invention in the preparation of drugs for preventing and treating ulcerative colitis is described in detail below in conjunction with the examples, but they should not be construed as limiting the scope of protection of the present invention.

[0038] Example 1

[0039] Experimental animals: 60 SPF-grade C57BL / 6 mice, male, 6-8 weeks old, weighing 18-24g, purchased from Qinglongshan Animal Breeding Farm in Jiangning District, Nanjing, and raised in an SPF-grade environment. Breeding environment: room temperature 24±2℃, humidity 50%-60%, 12h light / dark cycle. Mice were given free access to water and food, and the experimental grouping and modeling were carried out after 1 week of adaptive feeding.

[0040] Experimental groups: 60 C57BL / 6 mice were randomly divided into 5 groups, 10 mice in each group. Normal control group (Control group), model group (DSS group), sulfasalazine group (SASP group), low-dose avicularin group (L-Avicularin group), high-dose avicularin group (H-Avicularin group). SASP is a known anti-enteritis drug used as a positive control.

[0041] Establishment of UC mouse model: Except for the normal control group, the mice in other groups were given 3% DSS to prepare the UC model. The colitis model was prepared according to the literature method: 3% dextran sodium sulfate (DSS) solution was given to drink freely for 7 days, and then replaced with normal drinking water. After modeling, the activity status, coat color changes, body weight, stool characteristics and color, and the presence of bleeding of the mice were observed every day. The success of the model was evaluated based on the general condition of the mice, weight changes, disease activity index (DAI), etc. [Modeling method reference: Eichele DD, Kharbanda K K. Dextran sodium sulfate colitis murine model: An indispensable tool for advancing our understanding of inflammatory boweldiseases pathogenesis [J]. World journal of gastroenterology, 2017, 23 (33): 6016.]

[0042] Dosage scheme: After successful modeling, the model group, sulfasalazine group and scutellariae group were given corresponding interventions: the L-Avicularin group was given 25 mg / kg of Avicularin by gavage according to the weight of mice, the H-Avicularin group was given 50 mg / kg of Avicularin by gavage according to the weight of mice, the SASP group was given 1 mL / 100 g of SASP solution by gavage every day; the normal control group and the DSS model group were given ddH2O by gavage. The treatment lasted for 9 days, and the mice were killed by cervical dislocation under ether anesthesia on the 10th day, and serum and colon tissues were collected.

[0043] Detection indicators:

[0044] General condition: mental state, weight changes, feces characteristics, and DAI of mice (see Table 1 for scoring criteria).

[0045] Table 1 DAI scoring criteria

[0046] Weight loss / % Stool characteristics Occult blood in stool / visible blood in stool Scoring 0 normal normal 0 1-5 normal normal 1 5-10 Loose Occult blood positive 2 10-15 Loose Occult blood positive 3 >15 Loose stools Naked bloody stool 4

[0047] Colon injury evaluation indicators: colon length, colon pathological section HE staining, colon pathological score (scoring criteria see Table 2).

[0048] Table 2 Colon histopathological scoring criteria

[0049] score Lesion range Lesion range Lesion range Crypt destruction 0 none none none none 1 Submucosa Submucosa Submucosa Mild 2 Muscle layer Muscle layer Muscle layer Moderate 3 Serous layer Serous layer Serous layer Severe

[0050] Cytokine detection: ELISA and q-PCR techniques were used to detect the expression levels of interleukin 1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor α (TNF-α) in colon tissue.

[0051] Detection of signaling pathways and related proteins: Western Blot (WB) and immunofluorescence techniques were used to detect the expression levels of colon tight junction proteins ZO-1 and Occludin, as well as the expression levels of proteins related to the MAPK and NF-κB signaling pathways.

[0052] Experimental results:

[0053] Figure 1 A is the structural formula of Avicularin, B is the body weight test result, C is the blood stool result of mice, and D is the DAI index. Figure 1 It can be seen that the intake of Avicularin can significantly reduce the blood in stool of UC mice ( Figure 1 Middle C), reduce weight loss ( Figure 1 Middle B), reduce the DAI index ( Figure 1 (middle D). Figure 1 In the table, * compared with the DSS group, there was a statistically significant difference, p < 0.05; ** compared with the DSS group, there was a statistically significant difference, p < 0.01.

[0054] Figure 2 A is the observation result of mouse colon, B is the result of mouse colon length, C is the result of colon histopathology score, and D is the result of HE staining of colon pathological section. Figure 2 It can be seen that compared with the DSS group, the intake of Avicularin can significantly reduce the colon shortening of UC mice (see Figure 2 A and B), reducing pathological damage to colon tissue (see Figure 2 D), reducing colon pathology scores (see Figure 2 Middle C). Figure 2 **Compared with the DSS group, there was a statistically significant difference, p < 0.01.

[0055] Figure 3 A is the protein expression level of inflammatory cytokines IL-1β, IL-6, and TNF-α in each group, and B is the mRNA expression level. Figure 3 It can be seen that ELISA and q-PCR techniques were used to detect the expression levels of inflammatory cytokines IL-1β, IL-6, and TNF-α in colon tissue. Avicularin significantly reduced the expression levels of IL-1β, IL-6, and TNF-α proteins ( Figure 3 A) and mRNA expression levels ( Figure 3 In middle B), the difference was particularly obvious in the high-dose group. Figure 3In the table, * compared with the DSS group, there was a statistically significant difference, p < 0.05; ** compared with the DSS group, there was a statistically significant difference, p < 0.01.

[0056] Figure 4 The results of detecting the expression of tight junction proteins ZO-1 and occludin in colon tissue by WB and immunofluorescence were as follows ( Figure 4 A, B), the results showed that the intake of avicularin could significantly increase the expression levels of ZO-1 and occludin in UC mice, which to some extent showed that avicularin has a protective effect on the intestinal mucosal barrier. Figure 4 In the table, * compared with the DSS group, there was a statistically significant difference, p < 0.05, ** compared with the DSS group, there was a statistically significant difference, p < 0.01.

[0057] Figure 5 The results of the effect of avicularin on the MAPK / NF-κB signaling pathway were analyzed by WB. The results showed that avicularin had no significant effect on JNK and P38, but had a significant inhibitory effect on p-ERK ( Figure 5 In addition, avicularin significantly inhibited the expression of p-NF-κB and downstream proteins COX-2 and iNOS ( Figure 5 (middle B). Figure 5 In the table, * compared with the DSS group, there was a statistically significant difference, p < 0.05; ** compared with the DSS group, there was a statistically significant difference, p < 0.01.

[0058] In summary, the experimental results show that Avicularin can effectively improve DSS-induced ulcerative colitis in mice, not only improving colon damage and reducing the DAI index, but also reducing the expression of inflammatory cytokines and increasing the expression of intestinal mucosal barrier-related proteins. Its mechanism of action is related to the inhibition of the ERK / NF-κB signaling pathway.

[0059] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. Application of glycosides in the preparation of drugs for the prevention and treatment of ulcerative colitis.

2. The use according to claim 1, characterized in that: The ulcerative colitis is ulcerative colitis caused by dextran sulfate sodium.

3. The use according to claim 1, characterized in that: The glycoside exerts its effect by inhibiting the ERK / NF-κB signaling pathway.

4. The use according to claim 1, characterized in that: The dosage form of the drug for preventing and treating ulcerative colitis includes a suspension.

5. The use according to claim 4, characterized in that: When the dosage form of the drug for preventing and treating ulcerative colitis is a suspension, the preparation method of the drug for preventing and treating ulcerative colitis comprises the following steps: After dissolving scutellaria baicalensis in sodium carboxymethylcellulose solution, the solution is diluted with water to obtain the drug for preventing and treating ulcerative colitis.

6. The use according to claim 5, characterized in that: The mass concentration of the sodium carboxymethyl cellulose solution is 0.4-0.6%.

7. A drug for preventing and treating ulcerative colitis, characterized in that: The active ingredient is glycoside.

8. The drug according to claim 7, characterized in that Also includes auxiliary materials.

9. The drug according to claim 7 or 8, characterized in that The dosage form of the drug includes a suspension.

10. The use according to claim 9, characterized in that: When the dosage form of the drug is a suspension, the composition of the drug includes glycoside, sodium carboxymethyl cellulose and water.

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