本文關鍵詞：基于腦腸互動研究大黃治療大鼠腦出血的抗炎機制　出處：《南京中醫(yī)藥大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： 腦出血 腦腸互動 大黃 炎癥反應 CCK-8

【摘要】：目的:本研究運用實驗大鼠制造腦出血模型,選用中藥大黃作為處理因素,通過觀察檢測其對腦出血大鼠神經功能缺損、腦水腫程度、腸道動力功能、炎癥反應水平以及腦腸肽表達的干預作用,積極探討中藥大黃治療腦出血的抗炎機制。方法:將健康SD大鼠隨機分正常組、假手術組、模型組和大黃組四組,每組又分為24h、72h、120h三個時間點進行指標檢測。以大鼠作為研究模型,采用自體血注入法模擬大鼠自發(fā)腦出血。正常組正常飲食,無造模過程;假手術組,針體刺入但不注血,模型組和大黃組完成腦出血造模。大黃組按體重予生大黃濃煎液灌胃,余各組按體重使用蒸餾水灌胃,每天一次,持續(xù)5天。在相應時間點觀測神經功能行為學評分,采集記錄排便情況,采用Billiot公式測定腦組織含水量,使用多導生理記錄儀測定腸道動力,運用蛋白免疫印跡法、酶聯(lián)免疫吸附法、免疫組化法檢測炎癥反應水平及腦腸肽表達情況。實驗數據采用SPSS20.0統(tǒng)計軟件進行統(tǒng)計分析,多組間數據比較采用單因素方差分析(One-wayANOVA)進行統(tǒng)計處理,以P0.05作為差異有統(tǒng)計學意義的標準。結果:1.神經功能缺損評分:采用Longa評分法和平衡木評分法,正常組和假手術組各時間點評分為0,大黃組和模型組各時間點得分均較假手術組高,術后Oh評分最高,之后逐漸降低,造模后0h、24h兩組神經功能缺損評分無明顯差異,但在72h和120h的神經功能評分明顯低于模型組(P均0.05)。2.腸道動力功能:腦出血急性期24h內大鼠排便粒數較正常組組明顯減少(p0.01),大黃組排便粒數明顯多于模型組(p0.05)。各時間點正常組與假手術組腸段活力檢測值均無明顯差異(p0.05),各模型組較正常組腸段活力檢測值均有下降(p0.01),腦出血后24h模型組腸段活力檢測值最低,此后逐漸增加；大黃組72h、120h測得活力值均較模型組明顯增加(p0.05)。3.腦含水量:與假手術組相比,模型組各時間點腦組織含水量都有升高(p均0.01),72h達到高峰。隨時間延長,模型組和大黃組腦組織含水量均呈下降趨勢。與模型組相比,大黃組給藥后72h(p=0.043,p0.05)、120h(p0.01)腦組織含水量顯著減少,在120h時其含水量接進正常組組(p0.05)。4.腦組織、腸組織、血清炎癥反應水平:Western Blot檢測腦組織磷酸化NF-κB蛋白表達結果顯示,假手術組與正常組無明顯差異(p0.05),腦出血后24h模型組和大黃組腦組織表達量明顯高于假手術組,24、72h、120h大黃組明顯低于模型組(p均0.05)。ELISA檢測腦組織中TNF-α、IL-Iβ較假手術組明顯增高,24h大黃組較模型組IL-1β表達降低(p0.05),72h、120h大黃組較模型組TNF-α、IL-Iβ表達明顯降低(p0.01)。ELISA檢測腸組織模型組各時間點TNF-α、IL-1β表達水平較假手術組升高且差異有統(tǒng)計學差異(p0.01),24h大黃組較假手術組IL-1β表達降低(p0.05),72h、120h較模型組TNF-α、IL-1β表達明顯降低(p0.01)。ELISA檢測血清結果發(fā)現(xiàn),各時間點血清TNF-α、IL-1β表達水平較假手術組升高且有統(tǒng)計學差異(p0.01),大黃組與模型組各時間點比較均明顯降低(p0.05)。5.檢測發(fā)現(xiàn)24h、120h各組CCK-8表達無明顯差異,72h大黃組表達量較假手術組升高(p0.05),較模型組明顯升高(p0.05)。檢測腸組織CCK-8含量,發(fā)現(xiàn)72h大黃組較模型組明顯升高(p0.01),24h、120h無明顯差異。結論:1.中藥大黃能有效減輕腦出血后大鼠腦水腫程度,減輕神經損傷和促進神經功能恢復。2.中藥大黃能下調腦出血后大鼠腦組織NF-κB、TNF-α、IL-1β蛋白表達,減輕繼發(fā)性炎癥損傷。3.中藥大黃改善腦出血后大鼠腸道動力功能,下調腸道TNF-α、IL-1β蛋白表達。4.中藥大黃升高腸組織、腦組織腦腸肽CCK-8含量,對腦腸軸具有一定的調節(jié)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，

本文編號：1380394