逍遥散对焦虑模型大鼠胃黏膜HSP70及血清相关炎性细胞因子调节作用的研究

发布时间：2018-05-20 22:29

本文选题：焦虑症 + 胃黏膜　；参考：《北京中医药大学》2017年硕士论文

【摘要】：研究背景随着社会的进步、发展和不断变革,人们在享受着这一切带来的高效、便捷生活方式的同时,也承受着越来越大的压力。尤其是生活在城市的居民,要面临更多的工作、学习、择业等压力,面对更激烈的竞争,这时,人们往往更容易处于各种急、慢性应激状态。适度的应激可以提高机体的适应能力,使人保持上进心,但是过度的应激持续时间长,就会成为适应性的负担,久而久之就会导致机体焦虑。研究表明,长期处于过度应激状态易于导致焦虑,目前80%的疾病均与过度应激有密切的关系。长时间的持续应激,神经内分泌系统紊乱,引起相关炎性细胞因子的分泌、合成和转输异常。热休克蛋白70(HSP70)作为重要的保护蛋白,参与调控炎性细胞因子的表达。临床上的抗焦虑药物存在一定的副作用,研究者逐步把目光投向了中医学领域。研究目的本课题主要探讨逍遥散对慢性束缚应激焦虑模型大鼠胃黏膜HSP70及相关炎性细胞因子的调节作用,以亟探索逍遥散保护因情志不适而导致的胃黏膜损伤的作用机制,为临床应用逍遥散治疗因情志不适所导致的胃黏膜损伤提供实验依据。研究方法将48只健康的雄性SD大鼠(体重为220-250g)随机分为正常组、模型组、逍遥散组、咯利普兰组,每组分别12只。分组后的大鼠适应性喂养10d,实验组均以慢性束缚应激的方法制备焦虑大鼠模型:每天3h,连续14d。逍遥散组灌胃逍遥散悬浮液,而正常组和模型组分别灌胃去离子水,咯利普兰组注射相应药物。分别在第0、7、14d对大鼠进行体重测量、高架十字迷宫行为学测试;在实验的第0、14d做新环境进食抑制实验。每天定时观察大鼠的精神状态、身体姿势、皮毛色泽、活动度、对束缚的反应程度、眼黏膜色泽、耳廓色泽以及粪便等一般情况。通过免疫组化法检测胃黏膜HSP70的表达量。通过ELISA实验检测血清中IL-6、IL-10等相关炎性细胞因子的含量。结果1.一般状态,两周造模结束时,正常组、逍遥散组和咯利普兰组大鼠明显比模型组大鼠动作灵活,叫声柔和,嘶叫、反抗、挣扎的次数也明显较少,精神状态也尚可,毛发也比模型组整齐,粪便干湿适中呈球状;而模型组大鼠萎靡不振,毛发凌乱,色泽不明,蜷伏在角落里呈警惕状态,束缚时的反抗、嘶叫、挣扎也比一周前稍有减弱。2.体重变化,造模前1d及造模后第7d,各组大鼠体重未见明显差异,到第14d,与模型组相比,其余三组大鼠平均体重高于模型组,有统计学差异(P0.05),与正常组相比,逍遥散组及咯利普兰组大鼠体重无显著性差异。3.新环境进食抑制实验,在实验开始时,各组无统计学差异,而造模后的第14d,模型组时间明显多于逍遥散组。4.高架十字迷宫实验,两周造模实验结束后,与正常组相比,模型组大鼠封闭臂进入次数减少,停留时间延长;与正常组相比,其余三组双侧开放臂进入次数减少且停留时间减少,有统计学意义;与模型组相比,逍遥散组大鼠封闭臂进入次数增多且停留时间短,但均无统计学差异。5.免疫组化实验,模型组大鼠胃黏膜细胞表达的HSP70明显低于正常组,具有统计学意义(P0.05),与模型组大鼠相比,逍遥散组大鼠阳性表达量明显升高,且具有统计学意义(P0.01);逍遥散组和咯利普兰组间无统计学差异,逍遥散组表达量略高于咯利普兰组。6.ELISA实验,模型组大鼠血清中的IL-1β、IL-6、IL-10和TNF-α含量明显上升,高于其它三组,逍遥散组和咯利普兰组间无统计学意义。结论14d慢性束缚应激可以导致大鼠焦虑,逍遥散可通过促进胃黏膜产生大量HSP70的方式来抑制相关炎性细胞因子的释放,进而起到保护胃黏膜的作用,为临床应用逍遥散治疗因情志不适所导致的胃黏膜损伤提供实验依据。
[Abstract]:With the progress, development and continuous change of the society, people are enjoying more and more pressure while enjoying the high efficiency and convenient way of living. In particular, people living in the city should face more work, study, job selection, pressure, and more intense competition. At this time, people tend to be more easily located. In a variety of urgent, chronic stress state. Moderate stress can improve the adaptability of the body, keep the person into the heart, but excessive stress lasts long, it will become the burden of adaptability, and it will lead to the anxiety of the body for a long time. The study shows that the excessive stress is easy to cause anxiety for a long time, and 80% of the diseases are all over the time. Degree stress has a close relationship. Long lasting stress, neuroendocrine system disorder, secretion of related inflammatory cytokines, synthesis and transtransmission abnormalities. Heat shock protein 70 (HSP70) is an important protective protein and participates in the regulation of the expression of inflammatory cytokines. There are certain side effects in clinical anti anxiety drugs. The purpose of this study is to explore the effect of Xiaoyao Powder on the gastric mucosal HSP70 and related inflammatory cytokines in rats with chronic restraint stress anxiety, in order to explore the mechanism of the action of Xiaoyao Powder to protect the gastric mucosa caused by emotional discomfort. 48 healthy male SD rats (220-250g) were randomly divided into the normal group, the model group, the Xiaoyao San group, the rrrl group, and the group of 12. The rats were fed with 10d, and the experimental group was prepared with chronic restraint stress to prepare the anxiety rat model. Every day 3h, 14D. Xiaoyao Powder group was fed with Xiaoyao Powder, and the normal group and the model group were injected with deionized water, and the rrrl group was injected with the corresponding drugs. The rats were measured in 0,7,14d, the viaduct and the behavior test of the cross maze, and the new environmental feeding inhibition experiment was done in the 0,14d of the experiment. The general condition of mental state, body posture, skin color, activity, degree of reaction to bondage, eye mucous color, auricle color and feces. The expression of HSP70 in gastric mucosa was detected by immunohistochemical method. The content of inflammatory cytokines related to IL-6 and IL-10 in serum was detected by ELISA test. Results 1. general state and two weeks of modeling knot In the normal group, the rats in the normal group, the Xiaoyao group and the rlipulan group were more flexible than the model group, with a soft call, a scream, a rebellion, the struggle times were also less, the mental state was also clear, the hair was more neat than the model group, and the stool was moderately dry and wet. The model group was depressed, the hair was messy, the color and lustre was unidentified and curled up in the corner. In the state of vigilance, the resistance, the screaming, and the struggle were slightly weaker than the.2. weight before one week, and the weight of the rats in each group was not significantly different. The average weight of the other three groups of rats was higher than the model group, and the average weight of the other three groups was higher than that of the model group. The difference was statistically significant (P0.05), compared with the normal group, the Xiaoyao group and the normal group were compared with the normal group. There was no significant difference between the weight of the weight of the rat and the.3. new environment feeding inhibition experiment. At the beginning of the experiment, there was no statistical difference between each group. The time of the model group was significantly more than the.4. elevated cross maze test of the group after the model. After the two week model experiment, the number of closed arms in the model group was reduced and the number of closed arms decreased. Compared with the normal group, the number of open arms in the other three groups decreased and the retention time decreased. Compared with the model group, the entrance of the closed arm increased and the residence time was short, but there was no statistical difference between the three groups, but there was no statistical difference in the HSP70 immunohistochemistry test, and the expression of gastric mucosa cells in the model group was HSP70 bright. Significantly lower than the normal group, with statistical significance (P0.05), compared with the model group, the positive expression of the rats in the Xiaoyao group was significantly higher, and had statistical significance (P0.01). There was no statistical difference between the Xiaoyao Powder group and the rrrl group. The expression of Xiaoyao San group was slightly higher than the.6.ELISA test in the rrpulan group, and the IL-1 beta, IL in the serum of the model group of rats, IL The contents of -6, IL-10 and TNF- alpha were significantly higher than those of the other three groups, and there was no statistical significance between the Xiaoyao group and the ralprop group. Conclusion 14d chronic restraint stress can cause anxiety in rats. Xiaoyao San can inhibit the release of related inflammatory cytokines by promoting a large number of HSP70 in the gastric mucosa, and then play a role in protecting the gastric mucosa. To provide experimental evidence for clinical application of Xiaoyao San in treating gastric mucosal injury caused by emotional discomfort.
【学位授予单位】：北京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R285.5;R-332

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