基于脑肠互动研究大黄治疗大鼠脑出血的抗炎机制

发布时间：2018-01-04 22:26

  本文关键词：基于脑肠互动研究大黄治疗大鼠脑出血的抗炎机制　出处：《南京中医药大学》2017年硕士论文　论文类型：学位论文

  更多相关文章： 脑出血 脑肠互动 大黄 炎症反应 CCK-8

【摘要】：目的:本研究运用实验大鼠制造脑出血模型,选用中药大黄作为处理因素,通过观察检测其对脑出血大鼠神经功能缺损、脑水肿程度、肠道动力功能、炎症反应水平以及脑肠肽表达的干预作用,积极探讨中药大黄治疗脑出血的抗炎机制。方法:将健康SD大鼠随机分正常组、假手术组、模型组和大黄组四组,每组又分为24h、72h、120h三个时间点进行指标检测。以大鼠作为研究模型,采用自体血注入法模拟大鼠自发脑出血。正常组正常饮食,无造模过程;假手术组,针体刺入但不注血,模型组和大黄组完成脑出血造模。大黄组按体重予生大黄浓煎液灌胃,余各组按体重使用蒸馏水灌胃,每天一次,持续5天。在相应时间点观测神经功能行为学评分,采集记录排便情况,采用Billiot公式测定脑组织含水量,使用多导生理记录仪测定肠道动力,运用蛋白免疫印迹法、酶联免疫吸附法、免疫组化法检测炎症反应水平及脑肠肽表达情况。实验数据采用SPSS20.0统计软件进行统计分析,多组间数据比较采用单因素方差分析(One-wayANOVA)进行统计处理,以P0.05作为差异有统计学意义的标准。结果:1.神经功能缺损评分:采用Longa评分法和平衡木评分法,正常组和假手术组各时间点评分为0,大黄组和模型组各时间点得分均较假手术组高,术后Oh评分最高,之后逐渐降低,造模后0h、24h两组神经功能缺损评分无明显差异,但在72h和120h的神经功能评分明显低于模型组(P均0.05)。2.肠道动力功能:脑出血急性期24h内大鼠排便粒数较正常组组明显减少(p0.01),大黄组排便粒数明显多于模型组(p0.05)。各时间点正常组与假手术组肠段活力检测值均无明显差异(p0.05),各模型组较正常组肠段活力检测值均有下降(p0.01),脑出血后24h模型组肠段活力检测值最低,此后逐渐增加；大黄组72h、120h测得活力值均较模型组明显增加(p0.05)。3.脑含水量:与假手术组相比,模型组各时间点脑组织含水量都有升高(p均0.01),72h达到高峰。随时间延长,模型组和大黄组脑组织含水量均呈下降趋势。与模型组相比,大黄组给药后72h(p=0.043,p0.05)、120h(p0.01)脑组织含水量显著减少,在120h时其含水量接进正常组组(p0.05)。4.脑组织、肠组织、血清炎症反应水平:Western Blot检测脑组织磷酸化NF-κB蛋白表达结果显示,假手术组与正常组无明显差异(p0.05),脑出血后24h模型组和大黄组脑组织表达量明显高于假手术组,24、72h、120h大黄组明显低于模型组(p均0.05)。ELISA检测脑组织中TNF-α、IL-Iβ较假手术组明显增高,24h大黄组较模型组IL-1β表达降低(p0.05),72h、120h大黄组较模型组TNF-α、IL-Iβ表达明显降低(p0.01)。ELISA检测肠组织模型组各时间点TNF-α、IL-1β表达水平较假手术组升高且差异有统计学差异(p0.01),24h大黄组较假手术组IL-1β表达降低(p0.05),72h、120h较模型组TNF-α、IL-1β表达明显降低(p0.01)。ELISA检测血清结果发现,各时间点血清TNF-α、IL-1β表达水平较假手术组升高且有统计学差异(p0.01),大黄组与模型组各时间点比较均明显降低(p0.05)。5.检测发现24h、120h各组CCK-8表达无明显差异,72h大黄组表达量较假手术组升高(p0.05),较模型组明显升高(p0.05)。检测肠组织CCK-8含量,发现72h大黄组较模型组明显升高(p0.01),24h、120h无明显差异。结论:1.中药大黄能有效减轻脑出血后大鼠脑水肿程度,减轻神经损伤和促进神经功能恢复。2.中药大黄能下调脑出血后大鼠脑组织NF-κB、TNF-α、IL-1β蛋白表达,减轻继发性炎症损伤。3.中药大黄改善脑出血后大鼠肠道动力功能,下调肠道TNF-α、IL-1β蛋白表达。4.中药大黄升高肠组织、脑组织脑肠肽CCK-8含量,对脑肠轴具有一定的调节作用。
[Abstract]:Objective: This study used experimental rat model of intracerebral hemorrhage with manufacturing, rhubarb as processing factors, through the observation of the detection of neurological deficit in rats of cerebral hemorrhage, cerebral edema, intestinal motility, inflammatory reaction and intervention effect of expression of brain gut peptide, actively explore the anti-inflammatory mechanism of Rhubarb in the treatment of cerebral hemorrhage. Method: healthy SD rats were randomly divided into normal group, sham operation group, model group and rhubarb group four groups, each group was divided into 24h, 72h, 120h three time points were measured. The rats as the research model, the autologous blood in rats with spontaneous intracerebral hemorrhage model was injected into normal group with normal diet. No, modeling process; the sham operation group, the needle body piercing without blood injection molding, cerebral hemorrhage model group and rhubarb group. According to the weight to the rhubarb rhubarb group concentrated decoction orally, using distilled water according to the weight of other groups, once a day for 5 days . scores in the corresponding time points of observation of nerve function, recorded defecation, brain water content was determined by using the Billiot formula, the determination of intestinal motility using polygraph, using Western blot, enzyme-linked immunosorbent assay, immunohistochemical method to detect the inflammatory reaction and the level of brain gut peptide expression experiments. The data were analyzed by SPSS20.0 statistical software, data were compared with single factor analysis of variance (One-wayANOVA) statistical treatment, the difference was statistically significant with P0.05 as the standard. Results: 1. nerve function defect score: the Longa score and the balance score method, normal group and sham operation group at different time comment divided into 0 rhubarb group and the model group at each time point scores were higher than the sham operated group, postoperative Oh score was the highest, then gradually decreased after modeling, 0h, 24h two group of neural function defect score had no significant difference 寮，

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