免疫抑制剂SM934治疗银屑病的药理作用机制研究

发布时间：2018-05-13 17:09

本文选题：自身免疫疾病 + 免疫抑制剂　；参考：《中国科学院大学(中国科学院上海药物研究所)》2017年硕士论文

【摘要】：银屑病是一种常见的自身免疫疾病,临床表现为红色瘢痕,发病部位常遍布全身,严重影响患者生活1。目前为止银屑病的发病机制还不清楚,通常认为是由功能失调的角质细胞、活化的T细胞、固有免疫细胞以及多种炎症因子共同作用的结果。目前的治疗手段主要采取激素治疗、免疫抑制剂治疗、光疗法以及新兴的生物制剂疗法,但依然面临易复发的缺点,所以人们还在不断的努力寻找新的安全经济有效的治疗药物。青蒿素是从我国传统中药青蒿(Artemisia annua L.)中提取的有效成分,在治疗治疗疟疾方面有着高效低毒的优势。青蒿素所具备的广泛的抗炎抗肿瘤和免疫抑制的药理活性以及安全性,让它越来越受到科研人员的关注,但它同时也有着水溶性差,口服利用率低下的缺点,这限制了青蒿素的应用范围。本实验室对青蒿素进行了一系列结构改造和活性筛选,最终获得了多种青蒿素衍生物,其中SM934具备水溶性好,毒性低,活性高的优势,故将其作为候选药物,探索对银屑病的治疗作用。本课题的研究目的是通过体内外手段探究青蒿素衍生物SM934对银屑病的治疗作用。体内实验中,通过经皮给药的方式探索SM934对银屑病鼠尾鳞片模型的治疗作用,将BALB/c雌性小鼠作为实验动物,分别在鼠尾外涂SM934凝胶、维A酸、凝胶基质。2次/day,在第14天处死BALB/c小鼠,取距离尾根约2 cm处尾部背侧皮肤,HE染色,于显微镜下观察鳞片表皮是否有颗粒层形成,计算正角化率。结果显示SM934凝胶能有效地治疗鼠尾鳞片模型,促进角质形成细胞角质化。对于人永生化的角质形成细胞(HaCaT细胞),CCK8细胞毒性实验显示水溶性青蒿素衍生物SM934对HaCaT细胞毒性较低。在临床上,银屑病患者皮损处IL-6、IL-8、K17、S100A8的含量与皮损严重程度有一定的正相关关系。ELISA实验及RT-PCR的实验表明:TNF-α、IFN-γ共刺激能促进HaCaT细胞分泌细胞因子IL-6。TNF-α、IL-17共刺激能促进HaCaT细胞表达IL-6、IL-8、K17、S100A8、CXCL5等与银屑病发病相关的细胞因子。同时SM934能显著抑制HaCaT细胞表达上述细胞因子,抑制HaCaT细胞参与的炎症反应。体内试验显示,SM934能促进角质形成细胞正角质化,促进颗粒层细胞的形成,体外实验表明SM934能够抑制角质细胞炎症反应。说明SM934对于银屑病的有一定的治疗作用,为将来进一步探索青蒿素类衍生物治疗银屑病提供了实验依据。
[Abstract]:Psoriasis is a common autoimmune disease, clinical manifestations are red scar, the site of the disease is often spread throughout the body, seriously affecting the life of patients 1. So far, the pathogenesis of psoriasis is not clear. It is generally considered to be the result of the combined action of dysfunctional keratinocytes, activated T cells, innate immune cells and various inflammatory factors. The current treatments include hormone therapy, immunosuppressant therapy, phototherapy and emerging biologics, but they still face the drawback of being prone to relapse. So people are still trying to find new safe, economical and effective treatment drugs. Artemisia annua L. The active components extracted from the drug have the advantage of high efficiency and low toxicity in the treatment of malaria. Artemisinin has a wide range of anti-inflammatory, anti-tumor and immunosuppressive pharmacological activities and safety, making it more and more concerned by researchers, but it also has the shortcomings of poor water solubility and low oral utilization. This limits the application of artemisinin. A series of structural modification and activity screening of artemisinin were carried out in our laboratory, and a variety of artemisinin derivatives were obtained. Among them, SM934 has the advantages of good water solubility, low toxicity and high activity, so it is used as a candidate drug. To explore the therapeutic effect on psoriasis. The aim of this study was to explore the therapeutic effect of artemisinin derivative SM934 on psoriasis in vitro and in vivo. In vivo, the therapeutic effect of SM934 on the tail scales of psoriatic rats was explored by percutaneous administration. The female BALB/c mice were treated with SM934 gel and retinoic acid respectively. On the 14th day, the BALB/c mice were killed on the 14th day. The dorsal tail skin was stained with HE at a distance of about 2 cm from the tail root. The granular layer was observed under the microscope and the positive keratinization rate was calculated. The results showed that SM934 gel could effectively treat rat tail scale model and promote keratinization of keratinocytes. The cytotoxicity of SM934, a water-soluble artemisinin derivative, to HaCaT cells was found to be low in human immortalized keratinocytes. Clinically, In psoriatic patients, there was a positive correlation between the content of IL-6, IL-8, IL-8, S100A8, and the severity of the lesions. Elisa and RT-PCR experiments showed that the co-stimulation of TNF- 伪 and IFN- 纬 promoted the secretion of cytokine IL-6.TNF- 伪 -IL-17 by HaCaT cells, and promoted the expression of IL-6IL-8K17A100A8CXCL5 by HaCaT cells. The cytokines associated with the development of chiff disease. At the same time, SM934 could significantly inhibit the expression of cytokines in HaCaT cells and inhibit the inflammatory response of HaCaT cells. In vivo experiments showed that SM934 could promote the keratinization of keratinocytes and promote the formation of granular layer cells. In vitro, SM934 could inhibit the inflammatory response of keratinocytes. The results suggest that SM934 has a certain therapeutic effect on psoriasis and provides experimental basis for further exploration of artemisinin derivatives in the treatment of psoriasis.
【学位授予单位】：中国科学院大学(中国科学院上海药物研究所)
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R96

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