Use of aescin in the preparation of a drug for preventing and treating ulcerative colitis

Abstract

The application relates to the technical field of ulcerative colitis prevention and treatment, and particularly relates to application of avicularin in preparation of ulcerative colitis prevention and treatment drugs. Avicularin can effectively improve ulcerative colitis of mice caused by dextran sulfate sodium (DSS), can not only improve colon injury and reduce DAI index, but also can reduce expression of inflammatory cytokines and increase expression of intestinal mucosal barrier related proteins.

Description

Technical Field

[0001] This invention relates to the field of ulcerative colitis prevention and treatment technology, and in particular to the application of eurygmus glycoside in the preparation of drugs for the prevention and treatment of ulcerative colitis. Background Technology

[0002] Ulcerative colitis (UC) is a refractory disease characterized by abdominal pain, diarrhea, bloody and mucous stools, and tenesmus, which seriously affects patients' health and quality of life. Due to its difficulty in curing, high recurrence rate, and potential for malignant transformation, it has always been a hot topic in digestive health research. In recent years, with the improvement of living standards and changes in dietary structure and lifestyle, the incidence of ulcerative colitis in my country has been increasing year by year.

[0003] The goal of ulcerative colitis treatment is to rapidly induce and maintain remission, reduce hormone requirements, resistance, and dependence during treatment, prevent complications, and lower the surgical rate. Currently, commonly used Western medicine treatments include aminosalicylic acid preparations, immunosuppressants, hormones, and biological agents. While these have achieved some efficacy, their remission rates remain unsatisfactory. Furthermore, these drugs may have various side effects, even serious adverse reactions. Traditional Chinese medicine (TCM) has advantages such as high safety and fewer toxic side effects, with its effectiveness primarily stemming from its active ingredients. Reported applications of active TCM ingredients in the treatment of ulcerative colitis include alkaloids, flavonoids, saponins, and polysaccharides, specifically such as berberine, baicalin, puerarin, ginsenosides, total paeoniflorin, astragalus polysaccharides, and dandelion polysaccharides.

[0004] Avicularin is primarily derived from the whole herb of *Polygonum aviculare* L., a plant in the Polygonaceae family, but it is also found in many other plants. Avicularin is known to possess various biological activities, including anti-allergic, antioxidant, hepatoprotective, and antitumor effects. It has been reported to improve human hepatocellular carcinoma by regulating the activities of NF-κB (p65), cyclooxygenase-2 (COX-2), and peroxisome proliferation-activating receptor γ (PPAR-γ).

[0005] However, to date, there are no reports on the application of eurythrin as an anti-enteritis drug. Summary of the Invention

[0006] In view of this, the purpose of the present invention is to provide the application of euryptic glycoside in the preparation of drugs for the prevention and treatment of ulcerative colitis.

[0007] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0008] This invention provides the application of euryptoside in the preparation of drugs for the prevention and treatment of ulcerative colitis.

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

[0010] Preferably, the eurythrin 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 includes the following steps:

[0013] Polygonum aviculare was dissolved in sodium carboxymethyl cellulose solution and then 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] This invention provides a drug for the prevention and treatment of ulcerative colitis, the active ingredient of which is euryptoside.

[0016] Preferably, auxiliary materials are also included.

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

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

[0019] This invention provides the application of avicularin in the preparation of drugs for the prevention and treatment of ulcerative colitis. Avicularin can effectively improve ulcerative colitis in mice induced by sodium dextran sulfate (DSS), not only improving colonic 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. Attached Figure Description

[0020] Figure 1 The results of the prevention and treatment of DSS-induced ulcerative colitis by avicularin are shown. In the figure, A is the structural formula of avicularin, B is the weight test result, C is the mouse fecal blood test result, and D is the DAI index.

[0021] Figure 2 This is another part of the prevention and treatment results of Avicularin in DSS-induced ulcerative colitis, where A is the observation results of mouse colon, B is the result of mouse colon length, C is the result of colon histopathological scoring, and D is the result of HE staining of colon histopathological sections.

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

[0023] Figure 4 The expression results of tight junction proteins ZO-1 and occludin in colon tissue were detected by Western Blot (WB) and immunofluorescence. In the figure, A shows the protein expression levels of tight junction proteins ZO-1 and occludin in colon tissue as measured by WB, and B shows the results of tight junction proteins ZO-1 and occludin in colon tissue as detected by immunofluorescence.

[0024] Figure 5 The results show the effects of Avicularin on the MAPK / NF-κB signaling pathway, where A represents the results of Western blot detection of the MAPK signaling pathway, and B represents the results of Western blot detection of the NF-κB signaling pathway. Detailed Implementation

[0025] This invention provides the application of euryptoside in the preparation of drugs for the prevention and treatment of ulcerative colitis.

[0026] In this invention, the molecular formula of the eurythrin is C1 20 H 18 O 11 It can be obtained through purchase.

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

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

[0029] Avicularin can effectively improve ulcerative colitis in mice induced by sodium dextran sulfate (DSS). It can not only improve colonic 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 this 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 eurythrin in sodium carboxymethyl cellulose solution and then diluting it with water to obtain the drug for preventing and treating ulcerative colitis.

[0031] In this 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 this invention, the water is preferably double-distilled water (ddH2O).

[0033] This invention provides a drug for the prevention and treatment of ulcerative colitis, the active ingredient of which is euryptoside.

[0034] This invention does not impose any special requirements on the mass content of euryptoside in the drug. In this invention, based on the content of euryptoside in the drug, the preferred dosage of the drug is 25 mg / kg to 50 mg / kg mouse body weight.

[0035] In this invention, the drug for preventing and treating ulcerative colitis preferably also includes excipients. This invention does not have any special requirements on the type of excipients, and any medical excipients well known in the art are acceptable, such as excipients, antioxidants, etc.

[0036] In this invention, the dosage form of the drug preferably includes a suspension. When the drug is a suspension, the composition of the drug includes eurythrin, sodium carboxymethyl cellulose, and water.

[0037] The following examples illustrate the application of the eurythrin provided by the present invention in the preparation of drugs for the prevention and treatment of ulcerative colitis. However, these examples should not be construed as limiting the scope of protection of the present invention.

[0038] Example 1

[0039] Experimental animals: Sixty male SPF-grade C57BL / 6 mice, 6–8 weeks old, weighing 18–24 g, were purchased from Qinglongshan Animal Breeding Farm, Jiangning District, Nanjing City, and housed in an SPF-grade environment. Housing conditions: Room temperature 24±2℃, humidity 50%–60%, 12-hour light / dark cycle. Mice were given free access to water and food, and after one week of acclimatization, experimental grouping and modeling were initiated.

[0040] Experimental grouping: Sixty C57BL / 6 mice were randomly divided into 5 groups of 10 mice each. The groups were: normal control group (Control group), model group (DSS group), sulfasalazine group (SASP group), low-dose dextrin group (L-Avicularin group), and high-dose dextrin group (H-Avicularin group). SASP, a known anti-enteritis drug, was used as a positive control.

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

[0042] Dosage regimen: After successful model establishment in the model group, sulfasalazine group, and eurythrin group, corresponding interventions were administered: the L-Avicularin group received 25 mg / kg of Avicularin by gavage according to mouse body weight; the H-Avicularin group received 50 mg / kg of Avicularin by gavage according to mouse body weight; the SASP group received 1 mL / 100 g of SASP solution by gavage daily; the normal control group and DSS model group received ddH2O by gavage. The treatment continued for 9 days. On day 10, mice were euthanized under ether anesthesia with cervical dislocation, and serum and colon tissue were collected.

[0043] Testing indicators:

[0044] General conditions: mouse mental state, weight change, fecal characteristics, DAI (scoring criteria are shown in Table 1).

[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 stool Naked bloody stool 4

[0047] Evaluation indicators for colonic injury: colonic length, HE staining of colonic pathological sections, and colonic pathological score (scoring criteria are shown in Table 2).

[0048] Table 2. Scoring criteria for colonic histopathology

[0049] score lesion range lesion range lesion range crypt destruction 0 none none none none 1 Submucosal layer Submucosal layer Submucosal layer Mild 2 muscle layer muscle layer muscle layer moderate 3 Serum film layer Serum film layer Serum film layer Severe

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

[0051] Signaling pathway and related protein detection: Western blotting (WB) and immunofluorescence techniques were used to detect the expression levels of colonic 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 In the table, A represents the structural formula of Avicularin, B represents the body weight measurement result, C represents the mouse fecal blood test result, and D represents the DAI index. Figure 1 It can be seen that intake of Avicularin can significantly reduce fecal bleeding in UC mice. Figure 1 (C) Reduce weight loss ( Figure 1 (B) reduce the DAI index ( Figure 1 (D). Figure 1 Among them, * there was a statistically significant difference compared with the DSS group, p < 0.05; ** there was a statistically significant difference compared with the DSS group, p < 0.01.

[0054] Figure 2 Table A shows the observation results of the mouse colon, B shows the result of the mouse colon length, C shows the histopathological score of the colon, and D shows the result of HE staining of colon pathological sections. Figure 2 It can be seen that, compared with the DSS group, intake of Avicularin significantly reduced colonic shortening in UC mice (see...). Figure 2 (A and B), reducing pathological damage to colonic tissue (see A and B) Figure 2 (D), reduce colon pathology score (see D), reduce colon pathology score (see D) Figure 2 (C) Figure 2 In the study, there was a statistically significant difference between the **group and the DSS group (p < 0.01).

[0055] Figure 3 In the table, A represents the protein expression levels of the inflammatory cytokines IL-1β, IL-6, and TNF-α in each group, and B represents the mRNA expression levels. Figure 3 It was found that ELISA and q-PCR techniques detected the expression levels of inflammatory cytokines IL-1β, IL-6, and TNF-α in colon tissue, and Avicularin significantly reduced the protein expression levels of IL-1β, IL-6, and TNF-α. Figure 3 The expression levels of A and mRNA (in the middle) Figure 3 The differences were particularly pronounced in the medium- and high-dose groups (B). Figure 3Among them, * there was a statistically significant difference compared with the DSS group, p < 0.05; ** there was a statistically significant difference compared with the DSS group, p < 0.01.

[0056] Figure 4 The results of detecting the expression of tight junction proteins ZO-1 and occludin in colon tissue using Western blotting and immunofluorescence were presented. Figure 4 The results (A and B) showed that Avicularin intake significantly increased the expression levels of ZO-1 and occludin in UC mice, which to some extent indicates that Avicularin has a protective effect on the intestinal mucosal barrier. Figure 4 Among them, * showed a statistically significant difference compared with the DSS group (p < 0.05), and ** showed a statistically significant difference compared with the DSS group (p < 0.01).

[0057] Figure 5 To investigate the effects of avicularin on the MAPK / NF-κB signaling pathway, Western blotting was performed on the MAPK signaling pathway. The results showed that avicularin had no significant effect on JNK and p38, but it 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 (B) Figure 5 Among them, * there was a statistically significant difference compared with the DSS group, p < 0.05; ** there was a statistically significant difference compared with the DSS group, p < 0.01.

[0058] In summary, the experimental results indicate that Avicularin effectively improves DSS-induced ulcerative colitis in mice, not only improving colonic damage and reducing the DAI index, but also decreasing 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 description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of Polygonum aviculare as the sole active ingredient in the preparation of drugs for the prevention and treatment of ulcerative colitis.

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

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

4. The application according to claim 1, characterized in that, The dosage form of the drug for preventing and treating ulcerative colitis includes suspensions.

5. The application 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 includes the following steps: Polygonum aviculare was dissolved in sodium carboxymethyl cellulose solution and then diluted with water to obtain the drug for preventing and treating ulcerative colitis.

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

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