新疆紫草羟基萘醌对葡聚糖硫酸钠诱导的小鼠溃疡性结肠炎的治疗作用
发布时间：2018-09-19 10:31
【摘要】：炎性肠病(inflammatory bowel disease, IBD)，主要分为溃疡性结肠炎(ulcerative colitis, UC)和克罗恩病(Crohn’s disease，CD)，被世界卫生组织列为现代难治病之一，其发病率在世界范围内呈逐年增长趋势。IBD的发病机制至今尚未完全明确，可能的病因主要有感染、遗传、精神和免疫因素，其中免疫系统调节功能紊乱在IBD的发病机理中起着重要作用。目前IBD的治疗尚缺乏理想的药物，现有的治疗手段主要为药物缓解和手术切除，治疗药物包括：5-氨基水杨酸类、皮质类固醇类、免疫抑制剂和以肿瘤坏死因子-α（TNF-α）单抗为代表的生物制剂。然而，这些药物在治疗的过程中通常伴有多种毒副反应，，或出现药物抵抗或不能耐受的现象。如TNF-α单抗，除价格昂贵外，仍有一大部分对传统疗法抵抗的患者对该治疗方法无反应，因此，临床仍急需开发安全有效的新型治疗药物。 中药或天然产物毒副作用小、安全性好，从天然产物中寻找新的治疗药物一直是研究的热点。紫草羟基萘醌是从中国传统中药紫草中提取的一组化合物，是紫草中的主要抗炎活性成分，主要包含阿卡宁（alkannin）、紫草素（shikonin）及其衍生物。现代药理学研究表明，紫草羟基萘醌具有抗炎、促伤口愈合、抗溃疡和免疫调节活性，同时能够抑制TNF-α的转录激活，减少炎症介质的生成。但有关紫草羟基萘醌对炎性肠病的治疗作用尚未见报道。 我们从新疆紫草中提取得到一组羟基萘醌类混合物(hydroxynaphthoquinonemixture，HM)，结构研究表明：它主要含7种阿卡宁衍生物分别为阿卡宁、乙酰阿卡宁、β-乙酰氧基异戊酰阿卡宁、去氧阿卡宁、β,β′-二甲基丙烯酰阿卡宁、α-甲基丁酰阿卡宁、异戊酰阿卡宁。我们前期的研究表明：HM能够预防和治疗实验性关节炎，减轻三硝基苯磺酸(TNBS)诱导的结肠炎病变严重程度，降低炎症疾病模型大鼠血清和结肠组织中TNF-α含量。本课题采用葡聚糖硫酸钠(DSS)诱导的小鼠溃疡性结肠炎，进一步评价HM对IBD的治疗作用，并深入探讨其可能的作用机制。 本研究中，3.5%的DSS水溶液连续饮用6天成功诱导与人类UC类似的小鼠肠道炎症。自饮用DSS当天开始给药，小鼠灌胃给予HM (3.5、7、14mg/kg)和阳性对照药美沙拉嗪(200mg/kg)，每天给药一次，连续7天。结果显示，HM治疗可减轻小鼠溃疡性结肠炎病变严重程度，缓解动物体重下降，降低疾病活动指数评分，抑制结肠长度缩短。组织病理检查结果显示，HM各剂量组动物结肠组织病变程度较DSS模型组减轻，组织病理损伤评分显著降低，主要表现为病变范围、炎性细胞浸润减少，粘膜结构破坏程度减轻。此外，HM显著降低结肠组织中MPO活性和血清中TNF-α含量，其中中、高剂量组与DSS模型组比较，具有统计学显著性差异(P0.05)。Western blot实验结果显示，HM有效抑制结肠组织中TNF-α、p-IκBα和细胞核内NF-κB p65蛋白表达，上调IκBα蛋白表达，表明HM可能通过抑制TNF-α生成和NF-κB激活，阻断放大的炎症反应而发挥显著的抗炎效果。 以上研究结果表明，HM对DSS诱导的溃疡性结肠炎具有良好的治疗作用，它能够有效改善小鼠结肠炎临床症状和组织病理损伤，降低MPO活性和炎性细胞因子TNF-α含量，其作用机制可能与抑制TNF-α生成和NF-κB激活有关。研究发现为HM治疗IBD提供了药理学基础。
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R574.62
本文编号：2249878
[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.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R574.62
【参考文献】
相关期刊论文 前2条
1 甘华田,欧阳钦,陈友琴,夏庆杰;溃疡性结肠炎患者肠粘膜κ基因结合核因子的活化及抗炎药物的作用[J];中华医学杂志;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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