本文選題：胰性腦病　切入點：腦損傷　出處：《廣東藥學(xué)院》2014年碩士論文　論文類型：學(xué)位論文

【摘要】：【研究背景】胰性腦�。╬ancreatic encephalopathy, PE）是重癥急性胰腺炎（Severeacute pancreatitis, SAP）早期并發(fā)癥之一，是SAP病人出現(xiàn)中樞神經(jīng)系統(tǒng)（centralnervous system, CNS）癥狀，其發(fā)病率為18.2%，預(yù)后差，病死率高達(dá)67%，其主要臨床表現(xiàn)為定向力障礙、抑郁、意識模糊、煩躁、反應(yīng)遲鈍、表情淡漠等CNS癥狀。臨床上中藥制劑清胰湯用于治療SAP，使得SAP病人的癥狀明顯緩解，病程縮短，且減少并發(fā)癥。清胰顆粒（qingyi grains, QYG）即清胰湯方劑的固化制劑，進(jìn)一步探討QYG對SAP中腦損傷的作用機(jī)制及療效，有利于進(jìn)一步探索PE發(fā)病機(jī)制和研發(fā)QYG。 【目的】建立PE大鼠模型，探討在PE大鼠血腦屏障（blood brain barrier, BBB）通透性變化和腦組織中水通道蛋白-4（Aquaporin-4, AQP-4）的表達(dá)情況及相互關(guān)系，探討QYG治療后PE大鼠血腦屏障通透性變化和腦組織中水通道蛋白-4表達(dá)的影響，并探討QYG治療胰性腦病的相關(guān)保護(hù)機(jī)制，以其為QYG的臨床治療PE提供實驗依據(jù)。 【方法】90只健康成年的SD(Sprague-Dawley）大鼠，雌雄各半，隨機(jī)分成3組：假手術(shù)組(sham-operation, SO)組（30只）；PE組（30只）；QYG治療組(30只)。分別按12、24、48小時時間點處死老鼠10只。SO組逆行胰膽管行0.9%NaCl注射；PE組逆行胰膽管注射5%的牛黃膽酸鈉（sodium taurocholate, STC）(0.1ml/100g)建立PE模型組；QYG組建模后2H，QYG溶解以后灌胃，以后12h/次給藥，其它組則用0.9%NaCl代替QYG。（1）HE染色觀察胰腺病理變化；（2）伊文思藍(lán)測定血腦屏障通透性；（3）ELISA測定腦組織中TNF-α和AQP-4含量；（4）用免疫組化法來檢測腦組織中AQP-4蛋白表達(dá)情況；（5）定量逆轉(zhuǎn)錄聚合酶反應(yīng)（reverse transcription PCR, RT-PCR）測定腦組織中的AQP-4-mRNA表達(dá)的情況。 【結(jié)果】（1）PE組胰腺出現(xiàn)大量炎癥細(xì)胞浸潤，可觀察到鈣化灶，不同程度的出血、壞死，部分細(xì)胞出現(xiàn)核溶解或者消失，殘留的腺泡結(jié)構(gòu)出現(xiàn)腫脹，小葉間質(zhì)水腫，細(xì)胞極性消失；QYG組可觀察到腺泡結(jié)構(gòu)輕度腫脹，散發(fā)少量的充血及壞死，病變程度較PE組減輕，與PE組相比病理評分顯著降低（P0.05），PE組、QYG組與SO組相比均有病理評分均顯著升高（P0.05）。（2）QYG可能具有保護(hù)PE模型大鼠胰腺的作用。（3）PE組中BBB通透性從12h起開始升高，于24h達(dá)到峰值，，48h仍處于較高水平。與SO組相比，PE組、QYG組BBB通透性顯著升高（P0.05）；與PE組相比，QYG組BBB通透性顯著降低（P0.05）。（4）PE組和QYG組中腦組織TNF-α水平均高于SO組，PE組升高更明顯，在PE組中呈現(xiàn)時間遞增的關(guān)系，而QYG治療后TNF-α水平顯著降低（P0.05），QYG療效呈現(xiàn)時間依賴性。（5）大鼠腦組織中各組AQP-4表達(dá)水平不同，PE組AQP-412h開始升高，到48h達(dá)到峰值，QYG治療后腦組織AQP-4蛋白表達(dá)顯著降低，ELISA、免疫組化和QPCR結(jié)果變化趨勢相似。（6）經(jīng)統(tǒng)計相關(guān)分析腦組織中TNF-α與AQP-4存在正相關(guān)（r=0.932,P0.05），而BBB通透性與AQP-4呈現(xiàn)正相關(guān)（r=0.757,P0.05）。BBB通透性與腦組織中TNF-α呈現(xiàn)正相關(guān)（r=0.804,P0.05） 【結(jié)論】（1）大鼠PE模型建立成功，PE模型組中BBB通透顯著升高，至24h時達(dá)到頂峰，QYG治療后BBB通透性水平顯著降低。（2）QYG可能具有保護(hù)PE模型胰腺的作用。（3）PE模型組中腦組織的腫瘤壞死因子-α表達(dá)顯著升高，QYG治療后TNF-α水平顯著下降，說明QYG具有抗炎作用。（4）PE模型組中腦組織AQP-4蛋白表達(dá)顯著升高，而QYG治療后AQP-4表達(dá)水平下降，說明QYG對AQP-4介導(dǎo)的腦損傷具有保護(hù)作用。（5）PE模型組、QYG治療組中腦組織中TNF-α、AQP-4水平呈現(xiàn)逐漸升高的趨勢，與BBB通透性具有顯著相關(guān)，且存在正相關(guān)，說明TNF-α、AQP-4的升高可能導(dǎo)致BBB通透性增加，致使腦組織損傷。
[Abstract]:[background] pancreatic encephalopathy (pancreatic encephalopathy PE) is a severe acute pancreatitis (Severeacute pancreatitis SAP) is one of the early complications, SAP patients with central nervous system (centralnervous system CNS) symptoms, the incidence rate was 18.2%, poor prognosis and high fatality rate of 67%, the main clinical manifestations of orientation disorder, depression, confusion, irritability, unresponsive, indifferent expression and clinical symptoms of CNS. Traditional Chinese medicine Qingyi Decoction for the treatment of SAP, the SAP patient's symptoms, shorten the course of the disease, and reduce complications. Qingyi granule (Qingyi grains, QYG) - curable formulation of Qingyi Decoction prescription, further study the mechanism of QYG SAP brain injury and curative effect, help to further explore the pathogenesis and development of QYG. PE
[Objective] to establish a rat model of PE in PE rat blood brain barrier (blood brain, barrier, BBB) and the permeability of brain water channel protein -4 (Aquaporin-4, AQP-4) and the expression of the relationship, to explore the effects of QYG after treatment of water channel protein PE in rat blood brain barrier permeability and brain tissue changes the expression of -4, and to investigate the protective mechanism of QYG treatment of pancreatic encephalopathy, to provide experimental evidence for the clinical treatment of the PE QYG.
[Methods] 90 healthy adult SD rats (Sprague-Dawley), male and female, were randomly divided into 3 groups: sham operation group (sham-operation, SO) group (30 rats); group PE (30); QYG group (30). According to the time point of 12,24,48 mice were killed 10 hours in.SO group retrograde pancreatic duct injection of 0.9%NaCl; group PE by retrograde pancreatic duct injection of 5% sodium taurocholate (sodium taurocholate, STC) (0.1ml/100g) to establish PE model group; QYG after the establishment of model 2H, QYG dissolved after gavage, after 12h/ administration, the other group was replaced by 0.9%NaCl QYG. (1) to observe the pancreatic pathology the changes of HE staining; (2) blood brain barrier permeability by Evans blue; (3) determination of TNF- alpha and AQP-4 content in brain tissue of ELISA; (4) to detect the expression of AQP-4 protein in brain tissue by immunohistochemical method; (5) quantitative reverse transcription polymerase chain reaction (reverse transcription PCR, RT-PCR) in brain tissue in AQP- The expression of 4-mRNA.
銆愮粨鏋溿

本文編號：1570106