【摘要】：炎性腸病(inflammatory bowel disease, IBD)，主要分為潰瘍性結(jié)腸炎(ulcerative colitis, UC)和克羅恩病(Crohn’s disease，CD)，被世界衛(wèi)生組織列為現(xiàn)代難治病之一，其發(fā)病率在世界范圍內(nèi)呈逐年增長趨勢。IBD的發(fā)病機制至今尚未完全明確，可能的病因主要有感染、遺傳、精神和免疫因素，其中免疫系統(tǒng)調(diào)節(jié)功能紊亂在IBD的發(fā)病機理中起著重要作用。目前IBD的治療尚缺乏理想的藥物，現(xiàn)有的治療手段主要為藥物緩解和手術(shù)切除，治療藥物包括：5-氨基水楊酸類、皮質(zhì)類固醇類、免疫抑制劑和以腫瘤壞死因子-α（TNF-α）單抗為代表的生物制劑。然而，這些藥物在治療的過程中通常伴有多種毒副反應(yīng)，，或出現(xiàn)藥物抵抗或不能耐受的現(xiàn)象。如TNF-α單抗，除價格昂貴外，仍有一大部分對傳統(tǒng)療法抵抗的患者對該治療方法無反應(yīng)，因此，臨床仍急需開發(fā)安全有效的新型治療藥物。 中藥或天然產(chǎn)物毒副作用小、安全性好，從天然產(chǎn)物中尋找新的治療藥物一直是研究的熱點。紫草羥基萘醌是從中國傳統(tǒng)中藥紫草中提取的一組化合物，是紫草中的主要抗炎活性成分，主要包含阿卡寧（alkannin）、紫草素（shikonin）及其衍生物�，F(xiàn)代藥理學研究表明，紫草羥基萘醌具有抗炎、促傷口愈合、抗?jié)兒兔庖哒{(diào)節(jié)活性，同時能夠抑制TNF-α的轉(zhuǎn)錄激活，減少炎癥介質(zhì)的生成。但有關(guān)紫草羥基萘醌對炎性腸病的治療作用尚未見報道。 我們從新疆紫草中提取得到一組羥基萘醌類混合物(hydroxynaphthoquinonemixture，HM)，結(jié)構(gòu)研究表明：它主要含7種阿卡寧衍生物分別為阿卡寧、乙酰阿卡寧、β-乙酰氧基異戊酰阿卡寧、去氧阿卡寧、β,β′-二甲基丙烯酰阿卡寧、α-甲基丁酰阿卡寧、異戊酰阿卡寧。我們前期的研究表明：HM能夠預(yù)防和治療實驗性關(guān)節(jié)炎，減輕三硝基苯磺酸(TNBS)誘導的結(jié)腸炎病變嚴重程度，降低炎癥疾病模型大鼠血清和結(jié)腸組織中TNF-α含量。本課題采用葡聚糖硫酸鈉(DSS)誘導的小鼠潰瘍性結(jié)腸炎，進一步評價HM對IBD的治療作用，并深入探討其可能的作用機制。 本研究中，3.5%的DSS水溶液連續(xù)飲用6天成功誘導與人類UC類似的小鼠腸道炎癥。自飲用DSS當天開始給藥，小鼠灌胃給予HM (3.5、7、14mg/kg)和陽性對照藥美沙拉嗪(200mg/kg)，每天給藥一次，連續(xù)7天。結(jié)果顯示，HM治療可減輕小鼠潰瘍性結(jié)腸炎病變嚴重程度，緩解動物體重下降，降低疾病活動指數(shù)評分，抑制結(jié)腸長度縮短。組織病理檢查結(jié)果顯示，HM各劑量組動物結(jié)腸組織病變程度較DSS模型組減輕，組織病理損傷評分顯著降低，主要表現(xiàn)為病變范圍、炎性細胞浸潤減少，粘膜結(jié)構(gòu)破壞程度減輕。此外，HM顯著降低結(jié)腸組織中MPO活性和血清中TNF-α含量，其中中、高劑量組與DSS模型組比較，具有統(tǒng)計學顯著性差異(P0.05)。Western blot實驗結(jié)果顯示，HM有效抑制結(jié)腸組織中TNF-α、p-IκBα和細胞核內(nèi)NF-κB p65蛋白表達，上調(diào)IκBα蛋白表達，表明HM可能通過抑制TNF-α生成和NF-κB激活，阻斷放大的炎癥反應(yīng)而發(fā)揮顯著的抗炎效果。 以上研究結(jié)果表明，HM對DSS誘導的潰瘍性結(jié)腸炎具有良好的治療作用，它能夠有效改善小鼠結(jié)腸炎臨床癥狀和組織病理損傷，降低MPO活性和炎性細胞因子TNF-α含量，其作用機制可能與抑制TNF-α生成和NF-κB激活有關(guān)。研究發(fā)現(xiàn)為HM治療IBD提供了藥理學基礎(chǔ)。
[Abstract]:Inflammatory bowel disease (IBD), mainly divided into ulcerative colitis (UC) and Crohn's disease (CD), is listed as one of the modern refractory disease s by the World Health Organization. The incidence of IBD in the world is increasing year by year. The pathogenesis of IBD has not yet been fully defined, and the possible etiology is still unclear. There are mainly infection, heredity, psychiatry and immune factors, among which immune system dysfunction plays an important role in the pathogenesis of IBD. At present, there is no ideal drug for the treatment of IBD. The existing treatment methods are mainly drug relief and surgical resection. Therapeutic drugs include: 5-aminosalicylic acid, corticosteroids, immunosuppression. However, these drugs are usually accompanied by a variety of toxic and side effects, or drug resistance or intolerance. Therefore, it is still urgent to develop safe and effective new therapeutic drugs.
Traditional Chinese medicine or natural products have little side effects and good safety. It is always a hot spot to find new therapeutic drugs from natural products. Modern pharmacological studies have shown that Arnebia hydroxynaphthoquinone has anti-inflammatory, wound healing, anti-ulcer and immunomodulatory activities, and can inhibit the activation of TNF-a transcription and reduce the production of inflammatory mediators.
A group of hydroxynaphthoquinone mixtures (HM) were obtained from Arnebia sinensis. The structure studies showed that HM mainly contained seven kinds of Akanin derivatives, namely Akanin, AcetylAkanin, Beta-Acetyloxyisovaleryl Akanin, Deoxy-Akanin, Beta, Beta'-Dimethylacryloyl Akanin, and Alpha-Methylbutyryl Akanin. Ning, isovaloyl acanine. Our previous studies showed that HM can prevent and treat experimental arthritis, reduce the severity of TNBS-induced colitis, and reduce the content of TNF-a in serum and colon tissue of inflammatory disease model rats. This study used dextran sodium sulfate (DSS) induced ulcerative colon in mice. We further evaluated the therapeutic effect of HM on IBD and explored its possible mechanism.
In this study, 3.5% DSS solution was given to mice for 6 consecutive days to induce intestinal inflammation similar to human UC. The mice were given HM (3.5,7,14 mg/kg) and mesalazine (200 mg/kg) once a day for 7 consecutive days. The results showed that HM treatment could alleviate ulcerative colitis in mice. The results of histopathological examination showed that the degree of colonic lesion in HM group was less than that in DSS model group, and the score of histopathological lesion was significantly lower. The main manifestations were the extent of lesion, the infiltration of inflammatory cells and the destruction of mucosal structure. In addition, HM significantly decreased the activity of MPO in colon tissue and the content of TNF-alpha in serum, and there was a significant difference between the middle and high dose groups and DSS model group (P 0.05). Western blot results showed that HM effectively inhibited the expression of TNF-alpha, p-I kappa B alpha and NF-kappa B p65 protein in colon tissue, and up-regulated the expression of I kappa B alpha protein. HM may play a significant anti-inflammatory effect by inhibiting TNF-a production and NF-kappa B activation and blocking the amplified inflammatory response.
These results indicate that HM has a good therapeutic effect on DSS-induced ulcerative colitis. It can effectively improve the clinical symptoms and histopathological damage of mice colitis, reduce MPO activity and inflammatory cytokine TNF-alpha content. The mechanism may be related to the inhibition of TNF-alpha production and NF-kappa B activation. It provides the basis for pharmacology.
【學位授予單位】：吉林大學
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R574.62

【參考文獻】

相關(guān)期刊論文 前2條

1 甘華田,歐陽欽,陳友琴,夏慶杰;潰瘍性結(jié)腸炎患者腸粘膜κ基因結(jié)合核因子的活化及抗炎藥物的作用[J];中華醫(yī)學雜志;2002年06期

2 Praveen K Yadav;;Potential role of Th17 cells in the pathogenesis of inflammatory bowel disease[J];World Journal of Gastroenterology;2009年46期

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