大黄栓制剂对两种溃疡性结肠炎模型小鼠的作用研究

摘要/Abstract

摘要： 目的：构建2,4,6-三硝基苯磺酸（2,4,6-trinitrobenzene sulfonic acid，TNBS）和恶唑酮（oxazolone，OXZ）诱导的小鼠溃疡性结肠炎（ulcerative colitis，UC）模型，观察大黄栓剂对UC的治疗作用。方法：将420只昆明小鼠（♂）随机分为2组（n=210）制备小鼠TNBS和OXZ模型。Ⅰ组（TNBS模型组）将昆明小鼠（♂）随机分为7组（n=30）：A（TNBS模型组）、B（柳氮磺胺吡啶（Sulfasalazine，SASP）栓剂组）、C（大黄800 mg·kg-1栓剂组）、D（大黄400 mg·kg-1栓剂组）、E（大黄200 mg·kg-1栓剂组）、F（空白栓剂组）、G（TNBS溶剂对照组）。Ⅰ组（A~E）：0.6%TNBS溶液0.1mL灌肠；Ⅰ组（F）：不做任何处理；Ⅰ组（G）：50%乙醇0.1 mL灌肠；以上7组自灌肠一次后在d 1，d 2，d 3，d 5，d 7每组处死6只。Ⅱ组（OXZ模型组）将昆明小鼠（♂）随机分为7组（n=30）：A（OXZ模型组）、B（SASP栓剂组）、C（大黄800 mg·kg-1栓剂组）、D（大黄400 mg·kg-1栓剂组）、E（大黄200 mg·kg-1栓剂组）、F（空白栓剂组）、G（OXZ溶剂对照组）。Ⅱ组（A~E）：皮肤涂抹1% OXZ溶液（100%乙醇溶解）0.1mL每天一次，连续2 d（致敏），d 7以0.5% OXZ（50%乙醇溶解）0.1mL灌肠；Ⅱ组（F）：不做任何处理；Ⅱ组（G）：皮肤涂抹100%乙醇0.1mL，每天一次，连续2d，d 7以50%乙醇0.1mL灌肠；以上7组灌肠给药一次后在d 1~d 5每天处死6只小鼠。观察Ⅰ组，Ⅱ组小鼠疾病活动指数（disease activity index，DAI）、结肠组织大体损伤指数（colon macroscopic damage index，CMDI）和病理组织学评分（histopathological score，HPS），并检测小鼠结肠组织中髓过氧化物酶（myeloperoxidase，MPO）、白细胞介素-4（interleukin-4，IL-4）、肿瘤坏死因子-α （tumor necrosis factor-α，TNF-α）的水平。结果：Ⅰ组（A）小鼠的DAI、CMDI、HPS、MPO酶活性、IL-4含量、TNF-α表达含量与Ⅰ组（G）比较在d 1，d 2，d 3，d5，d7均有明显改变；Ⅰ组给予含药栓剂组（B~E）与Ⅰ组空白栓剂组（F）相比，小鼠的DAI、CMDI、HPS、MPO酶活性、IL-4含量、TNF-α表达在d 1，d 2，d 3，d5，d7均有差异；Ⅰ组大黄不同剂量组（C~E）与Ⅰ组SASP栓剂组（B）相比，在d 1，d 2，d 3，d5，d7均有变化，其中大黄200 mg·kg-1栓剂组（E）差异较小。Ⅱ组（A）小鼠的DAI、CMDI、HPS、MPO酶活性、IL-4含量、TNF-α表达含量与Ⅱ组（G）比较在d 1~d 5均有显著性差异；Ⅱ组给予含药栓剂组（B~E）与Ⅱ组（F）相比在d 1~d 5均有变化，其中大黄给药组（C~E）有显著性差异；Ⅱ组大黄不同剂量组（C~E）与Ⅱ组SASP栓剂组（B）相比在d 1~d 5均有变化，其中大黄200 mg·kg-1栓剂组（E）差异较小。结论：TNBS与OXZ均能成功诱导小鼠UC模型；大黄对小鼠UC有一定的治疗作用，以大黄200 mg·kg-1剂量疗效较佳，但是不及SASP效果。

关键词: 溃疡性结肠炎, 2,4,6-三硝基苯磺酸, 恶唑酮, 大黄, 栓剂

Abstract: Objective: To construct ulcerative colitis (UC) model included by 2,4,6- trinitrobenzene sulfonic acid (TNBS) and oxazolone (OXZ) in mice and to observe the therapeutic effect of rhubarb suppository on ulcerative colitis. Methods：448 KM mice (♂) were randomized into 2 groups (n = 224) to construct TNBS and OXZ model. Kunming Mice in GroupⅠ(TNBS model group) were randomly divided into 7 subgroups (n=32) including A (TNBS model group), B (sulfasalazine (SASP) suppository group), C (800 mg·kg-1 rhubarb suppository group), D (400 mg·kg-1 rhubarb suppository group), E (200 mg·kg-1 rhubarb suppository group), F (blank suppository group) and G (TNBS solvent control group) . GroupⅠ(A ~ E) were treated with 0.6% TNBS solution 0.1 mL enema; GroupⅠ(F) had no treatment; GroupⅠ(G) with 50% ethanol 0.1 mL enema; 6 mice of each group were sacrificed after giving enema on d 1, d 2, d 3, d 5, and d 7. Mice in GroupⅡ(OXZ model group♂) were randomly divided into 7 subgroups (n=32) including A (TNBS model group), B (sulfasalazine (SASP) suppository group), C (800 mg·kg-1 rhubarb suppository group), D (400 mg·kg-1 rhubarb suppository group), E (200 mg·kg-1 rhubarb suppository group), F (blank suppository group) and G (TNBS solvent control group) . GroupⅡ(A~E) were treated with skin sensitization 1% OXZ solution (0.1 mL 100% ethanol solution) once a day, lasting two days (sensitization) and on the 7th day they were treated with OXZ at 0.5% (50% ethanol solution) 0.1 mL enema; GroupⅡ(F) had no treatment; GroupⅡ(G) with skin sensitization 100% ethanol 0.1 mL, once a day, lasting two days, and on the 7th day with 0.1 mL of 50% ethanol enema; 6 mice of each group were sacrificed after giving enema on the 1st, 2nd, 3rd, 4th, 5th day. We observed the Disease Activity Index (DAI), Colon Macroscopic Damage Index (CMDI) and Histopathological Score (HPS) score of the mice in GroupⅠand GroupⅡ, and determined the levels of Myeloperoxidase (MPO), Interleukin -4 ( IL-4) and Tumor Necrosis Factor -α (TNF-α) in colon tissues of mice. Results：Compared with GroupⅠ(G), the activity of DAI, CMDI, HPS and MPO, IL-4 content and TNF-α expression content in GroupⅠ(A) changed significantly on d 1, d 2, d 3, d 5, and d 7. In GroupⅠ, compared with blank suppository group (F) , the DAI, CMDI, HPS, MPO activity, IL-4 content, and TNF-α expression content of the mice in groups (B~E) had differences on d 1, d 2, d 3, d 5, d 7. Compared with SASP suppository group (B) in GroupⅠ, there were differences in (C~E) groups with different rhubarb doses on d 1, d 2, d 3, d 5, and d 7, in which the rhubarb 200 mg·kg-1 suppository group (E) had little difference from other rhubarb suppository groups. Compared with groupⅡ(G), the activity of DAI, CMDI, HPS and MPO , IL-4 content and TNF-αexpression content of the mice in GroupⅡ(A) had significant difference on d 1~5. Compared with blank suppository group (F) in GroupⅡ, the activity of DAI, CMDI, HPS and MPO, IL-4 content, and TNF-α expression content in mice of suppository group (B~E) took changes on d 1~5, and had significant changes in groups (C~E) with rhubarb. In GroupⅡ, compared with SASP suppository group (B), groups (C~E) with different doses of rhubarb had differences, in which rhubarb 200 mg·kg-1 suppository group (E) had little difference. Conclusion：Both TNBS and OXZ could induce mice model of ulcerative colitis. Rhubarb with different doses performed therapeutic effects on UC mice, and rhubarb with 200 mg·kg-1 dose did better, but not as good as the effect of SASP .

Key words: ulcerative colitis, 2,4,6-trinitrobenzene sulfonic acid, Oxazolone, , rhubarb, suppository

中图分类号:

R965

张丹参，王倩，田慧，薛贵平，张力. 大黄栓制剂对两种溃疡性结肠炎模型小鼠的作用研究[J]. 神经药理学报, 2015, 5(1): 19-37.

ZHANG Dan-shen，WANG Qian，TIAN Hui，XUE Gui-ping，ZHANG Li. Effects of Rhubarb Suppository on Two Kinds of Ulcerative Colitis Model in Mice[J]. Acta Neuropharmacologica, 2015, 5(1): 19-37.

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