4-氨基水杨酸和5-氨基水杨酸抗炎作用及作用机制研究

发布时间：2018-05-16 19:24

本文选题：溃疡性结肠炎 + 4-ASA　；参考：《山西医科大学》2014年硕士论文

【摘要】：溃疡性结肠炎（ulcerative colitis，UC）是一种反复发作的慢性的非特异性结、直肠炎，以腹痛、腹泻、里急后重、粘液脓血便等为主要临床症状[1-3]。该病起病缓慢，病程迁延不愈，长达几年甚至数十年，症状易反复发作，是胃肠道中除恶变外最严重的疾病之一。目前，药物治疗仍然是UC的主要治疗手段。其中，5-氨基水杨酸（5-aminosalicylic acid，5-ASA）及其前药是治疗轻、中度广泛性UC的一线药物。近年来，许多实验研究证实4-氨基水杨酸（4-aminosalicylic acid，4-ASA）在UC的治疗上与5-ASA有相近的疗效，已用于不能耐受柳氮磺胺吡啶（sulfasalazine，SASP）的轻、中度UC。本论文由三章组成，分别从动物水平和细胞水平研究4-ASA和5-ASA的抗炎作用，并对两者发挥抗炎作用的作用机制进行了研究。第一章采用TNBS法诱导大鼠溃疡性结肠炎模型后，分别设正常组、模型组、SASP组、4-ASA组和5-ASA组，观察4-ASA和5-ASA对大鼠UC的治疗作用。实验结果显示：模型组大鼠疾病活动指数评分、大体评分和组织学评分均较对照组明显增高，而造模后给予SASP、4-ASA、5-ASA后，各组大鼠结肠炎症程度明显减轻，，DAI评分、大体评分、组织学评分均较模型组大鼠显著减低（P＜0.05）。第二章采用脂多糖（lipopolysaccharide，LPS）诱导小鼠单核巨噬细胞（RAW264.7）炎症后，考察低（10μg/ml）、中（100μg/ml）、高（1000μg/ml）浓度4-ASA和5-ASA对炎性细胞IL-6、NO、iNOSmRNA及iNOS蛋白表达的抑制作用。实验结果显示：4-ASA：与正常组比较，模型组细胞内IL-6、NO、iNOSmRNA及iNOS蛋白表达表达均显著上调（P＜0.05）。与模型组相比，造模后给予10μg/ml4-ASA处理组的细胞内IL-6、iNOSmRNA及iNOS蛋白表达未见明显下调（P＞0.05），NO表达明显下调（P＜0.05）。造模后给予100μg/ml和1000μg/ml4-ASA处理组的细胞内IL-6、NO、iNOSmRNA及iNOS蛋白表达均明显下调（P＜0.05）。5-ASA：与正常组比较，模型组细胞内IL-6、NO、iNOSmRNA及iNOS蛋白表达表达均显著上调（P＜0.05）。与模型组相比，造模后给予10、100和1000μg/ml5-ASA处理组的细胞内IL-6、NO、iNOSmRNA及iNOS蛋白表达均明显下调（P＜0.05）。第三章采用脂多糖（lipopolysaccharide，LPS）诱导小鼠单核巨噬细胞（RAW264.7）炎症后，考察低（10μg/ml）、中（100μg/ml）、高（1000μg/ml）浓度4-ASA和5-ASA对炎性细胞JNK1/2、p38MAPK磷酸化及IκBα表达的抑制作用。实验结果显示：4-ASA：与正常组比较，模型组细胞内p-JNK1/2、p-p38及IκBα蛋白表达均显著上调（P＜0.05）。与模型组相比，造模后给予10和100μg/ml4-ASA处理组的细胞内p-JNK1/2蛋白表达未见明显下调（P＞0.05），1000μg/ml4-ASA处理组的细胞内p-JNK1/2蛋白表达明显下调（P＜0.05）。造模后给予10、100和1000μg/ml4-ASA处理组的细胞内p-p38和IκBα蛋白表达均未见显著下调（P＞0.05）。5-ASA：与正常组比较，模型组细胞内p-JNK1/2、p-p38及IκBα蛋白表达均显著上调（P＜0.05）。与模型组相比，造模后给予10和100μg/ml5-ASA处理组的细胞内p-JNK1/2蛋白表达未见明显下调（P＞0.05），1000μg/ml5-ASA处理组的细胞内p-JNK1/2蛋白表达明显下调（P＜0.05）。造模后给予10、100和1000μg/ml4-ASA处理组的细胞内p-p38蛋白表达均显著下调（P＜0.05），IκBα蛋白表达均未见显著下调（P＞0.05）。由以上三部分实验结果可以得出结论：4-ASA和5-ASA均具有明显抗炎作用，但二者发挥抗炎作用的机制有所不同。4-ASA主要通过抑制JNK1/2的磷酸化而发挥作用，而5-ASA同时抑制JNK1/2和p38的磷酸化而发挥作用。
[Abstract]:Ulcerative colitis (ulcerative colitis, UC) is a chronic, chronic, nonspecific knot, proctitis, with abdominal pain, diarrhea, heavy weight in the back, mucus and blood and urine as the main clinical symptoms, [1-3]., the disease is slow, the course of the disease is prolonged for several years or even ten years, and the symptoms are prone to relapse, which is the most serious outside of the gastrointestinal tract. At present, drug therapy is still the main treatment for UC. Among them, 5- amino salicylic acid (5-aminosalicylic acid, 5-ASA) and their prodrugs are the first-line drugs for the treatment of mild, moderate and extensive UC. In recent years, many experimental studies have confirmed that 4- amino salicylic acid (4-aminosalicylic acid, 4-ASA) is similar to 5-ASA in the treatment of UC. It has been used for the light and moderate UC., which is not tolerable to sulfasalazine (SASP). This paper is composed of three chapters. The anti-inflammatory effects of 4-ASA and 5-ASA are studied from animal level and cell level, and the mechanism of anti-inflammatory action of both of them has been studied.
In the first chapter, the rat model of ulcerative colitis was induced by TNBS, and the normal group, model group, SASP group, 4-ASA group and 5-ASA group were set up to observe the therapeutic effect of 4-ASA and 5-ASA on UC in rats. The results showed that the score of disease activity index, gross score and histological score of the model group were significantly higher than those of the control group, and the model was given after the model group. After SASP, 4-ASA, 5-ASA, the degree of colitis in each group was significantly reduced, DAI score, gross score and histological score were significantly lower than those in the model group (P < 0.05).
In the second chapter, after lipopolysaccharide (LPS) was used to induce mononuclear macrophage (RAW264.7) inflammation in mice, the inhibitory effects of low (10 g/ml), medium (100 u g/ml), and high (1000 mu) concentration of 4-ASA and 5-ASA on the expression of IL-6, NO, iNOSmRNA and iNOS protein in inflammatory cells were investigated. The expression of IL-6, NO, iNOSmRNA and iNOS increased significantly (P < 0.05). Compared with the model group, the expression of IL-6, iNOSmRNA and iNOS protein was not significantly down (P > 0.05) and NO expression decreased significantly (P < 0.05) after the model group was given to the 10 micron g/ml4-ASA treatment group. The expression of NOSmRNA and iNOS protein decreased significantly (P < 0.05).5-ASA. Compared with the normal group, the expression of IL-6, NO, iNOSmRNA and iNOS protein in the cells of the model group were significantly up (P < 0.05). Compared with the model group, the IL-6 in the cells of the 10100 and 1000 mu g/ml5-ASA treated groups was significantly down (0). .05).
In the third chapter, after lipopolysaccharide (LPS) was used to induce mononuclear macrophage (RAW264.7) inflammation in mice, the inhibitory effects of low (10 g/ml), medium (100) g/ml, and high (1000 mu) concentration of 4-ASA and 5-ASA on inflammatory cells JNK1/2, p38MAPK phosphorylation and I B alpha expression were investigated. Experimental results showed that the model group cells were compared with the normal group. The expression of p-JNK1/2, p-p38 and I kappa B alpha protein was significantly up-regulated (P < 0.05). Compared with the model group, the expression of p-JNK1/2 protein in the cells of 10 and 100 mu g/ml4-ASA treated groups was not significantly down (P > 0.05). The intracellular p-JNK1/2 protein table of the 1000 UU treatment group was down significantly down (P < 0.05). 10100 and 1000 micron were given after the model group. The expression of p-p38 and I kappa B alpha in the l4-ASA treatment group did not decrease significantly (P > 0.05).5-ASA: compared with the normal group, the expression of p-JNK1/2, p-p38 and I kappa B alpha protein in the cells of the model group was significantly up (P < 0.05). Compared with the model group, the expression of intracellular protein in the 10 and 100 micron groups was not obvious after the model group. Down regulation (P > 0.05), the expression of intracellular p-JNK1/2 protein in the 1000 g/ml5-ASA treatment group decreased significantly (P < 0.05). The expression of p-p38 protein in the cells of the treatment group was significantly down regulated (P < 0.05), and the expression of I kappa B alpha protein was not significantly down (P > 0.05).
From the experimental results of the above three parts, we can conclude that both 4-ASA and 5-ASA have obvious anti-inflammatory effects, but the mechanisms of the two anti inflammatory effects are different by inhibiting the phosphorylation of JNK1/2, while 5-ASA inhibits the phosphorylation of JNK1/2 and p38 at the same time.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R574.62

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