發(fā)布時間：2018-04-16 07:46

  本文選題：SAH + 縫隙連接�。� 參考：《南昌大學(xué)》2014年碩士論文

【摘要】：目的： 自發(fā)性蛛網(wǎng)膜下腔出血（SAH）占腦血管意外的5%左右，在腦血管意外中僅低于腦內(nèi)出血和腦梗死的發(fā)病率，但SAH的病死率明顯高于后兩者[1]，并且目前缺乏特效治療措施。SAH后出現(xiàn)的遲發(fā)性缺血性神經(jīng)功能障礙（DIND）是導(dǎo)致高致死高致殘率的主要原因。DIND一直是神經(jīng)外科學(xué)者研究的重點和熱點。DIND的致病因素，公認(rèn)的主要的兩個理論：即大血管的延遲性痙攣，及微血管的功能紊亂。我們實驗室前期研究表明：縫隙連接（GJ）可能在自發(fā)性蛛網(wǎng)膜下腔出血后的DIND病理過程中起到十分重要的作用。應(yīng)用特異性的Cx43siRNA可有效地沉默縫隙連接蛋白43（Cx43）的表達(dá)水平，同時能減輕腦血管痙攣的程度[40]。在前期研究的基礎(chǔ)上，本實驗擬驗證：有效下調(diào)縫隙連接Cx43后對SAH后發(fā)生的DIND的影響。 方法： 1、二次注血模型的建立：采用普通級SD大鼠，腹腔麻醉后抽取尾靜脈血約0.3ml備用，體式顯微鏡下枕大池穿刺，見清亮腦脊液流出約0.1ml，將尾靜脈血注入，次日進(jìn)行同前步驟，完成整個造模過程。 2、SAH模型的鑒定：分為單純SAH造模組和正常組（假手術(shù)組），第七天應(yīng)用墨汁染色法測量基底動脈直徑及神經(jīng)功能評分評價神經(jīng)功能缺損，鑒定SAH模型。 3、病毒的沉默效率檢測：采用腺病毒枕大池注射設(shè)立病毒處理組，單純SAH組和正常組對照。分1d，3d，7d，14d四個時間點處死SD大鼠，取基底動脈，Western bloting檢測病毒沉默Cx43的效率。 4、神經(jīng)功能評分：單純SAH造模組和正常組，預(yù)處理組，處理組等分7d，14d兩個時間行神經(jīng)功能評分（改良的garcia JH和beam balance評分法）。 5、MRI檢測：正常組和單純SAH造模組，預(yù)處理組，處理組分7d，14d兩個時間點行MRI（DWI，PWI）檢測，，包括腦血流量（CBF），表觀彌散系數(shù)（ADC）值的檢測。 結(jié)果： 1、印度墨汁染色法檢測基底動脈直徑，造模后7天，單純SAH造模組的基底動脈直徑明顯小于假手術(shù)對照組。 2、腺病毒沉默效率檢測：腺病毒腦池注射可以有效的沉默腦血管的Cx43表達(dá)。 3、神經(jīng)功能評分：改良的garcia評分法：SAH組與正常組比，7d，出現(xiàn)神經(jīng)功能缺損癥狀，14天神經(jīng)功能缺損癥狀與正常組比無明顯差異；所有的實驗處理組與單純SAH組比較，均不存在明顯的神經(jīng)功能缺損；Beam balance評分法，7d，14d所有組別之間不存在明顯差異。 4、MRI結(jié)果：SAH組與正常組比較，7d出現(xiàn)CBF及ADC值的下降；病毒預(yù)處理組和病毒處理組與單純的SAH造模組比，7d可以改善CBF，14天未能改善CBF，7d，14d均未能改善ADC值；甘珀酸預(yù)處理、甘珀酸處理組、病毒溶媒預(yù)處理組、病毒溶媒處理組、甘珀酸溶媒預(yù)處理組及甘珀酸溶媒處理組與單純SAH組比較，在7d和14d兩個時間點未能改善CBF，ADC值。 結(jié)論： 1、自體尾靜脈血二次枕大池注血模型能夠再現(xiàn)SAH的病程。 2、枕大池注射腺病毒，特異性下調(diào)Cx43后可以改善SAH后的CBF，提示 能夠改善SAH的預(yù)后。
[Abstract]:Objective:Spontaneous subarachnoid hemorrhage (SAH) accounts for about 5% of cerebrovascular accidents, and is only lower than the incidence of intracerebral hemorrhage and cerebral infarction in cerebrovascular accidents.However, the fatality rate of SAH is significantly higher than that of the latter two [1], and the lack of special therapeutic measures. The delayed ischemic neurological dysfunction (DINDD) is the main cause of high mortality and high disability rate. Dind has always been the research of neurosurgery scholars.Focus and focus on the pathogenic factors of .DIND,Two major theories are generally accepted: delayed vasospasm of large vessels and dysfunction of microvasculature.Our previous laboratory studies have shown that gap junction (GJ) may play an important role in the pathological process of DIND after spontaneous subarachnoid hemorrhage.The application of specific Cx43siRNA can effectively silence the expression of gap junction protein 43 (Cx43) and reduce the degree of cerebral vasospasm [40].On the basis of previous studies, this experiment is intended to verify the effect of down-regulation of gap junction Cx43 on DIND after SAH.Methods:Establishment of blood injection model: caudal venous blood was extracted from normal SD rats after abdominal anaesthesia about 0.3ml was used, occipital cistern puncture was performed under the posture microscope, clear cerebrospinal fluid (CSF) flowed out about 0.1ml, caudal vein blood was injected into caudal vein blood, and the same steps were performed the next day.Complete the whole modeling process.2Identification of SAH model: it was divided into simple SAH model group and normal group (sham-operation group). On the 7th day, the diameter of basilar artery and nerve function score were measured with ink staining method to evaluate the nerve function defect, and the SAH model was identified.3. Detection of virus silencing efficiency: adenovirus cistern cistern injection was used to set up virus treatment group, SAH group and normal control group.Sprague-Dawley rats were killed at four time points (1 d, 3 d, 7 d, 14 d). Western bloting was taken from basilar artery to detect the efficiency of virus silencing Cx43.4. Neurological function score: SAH model group and normal group, preconditioning group and treatment group were divided into 7 days and 14 days to perform nerve function score (modified garcia JH and beam balance scoring method).5MRI examination: normal group and simple SAH model group, preconditioning group and treatment group were examined with MRII-DWII-PWI at 14 days after 7 days, including cerebral blood flow (CBF), apparent diffusion coefficient (ADCC).Results:1. The diameter of basilar artery was detected by Indian ink staining. The diameter of basilar artery in SAH group was significantly smaller than that in sham operation group 7 days after modeling.2, detection of adenovirus silencing efficiency: adenovirus cistern injection can effectively silence the expression of cerebrovascular Cx43.3, nerve function score: the improved garcia score method: compared with the normal group, there was no significant difference between the control group and the control group in 14 days after the onset of neurologic deficit symptoms, all the experimental treatment groups were compared with the simple SAH group, and all the experimental treatment groups were compared with the simple SAH group.There was no significant difference between all groups in 7 days and 14 days.(4) the results of MRI showed that the CBF and ADC decreased on the 7th day compared with the normal group, the virus pretreatment group and virus treatment group could improve the ADC value after 14 days compared with the simple SAH model group, and the glycyroxic acid pretreatment group and the glycyrrhizic acid treatment group could not improve the ADC value after 14 days.Compared with SAH group, virus solvent pretreatment group, virus solvent pretreatment group, glycinic acid solvent pretreatment group and glycinic acid solute pretreatment group could not improve the SAH value at 7 and 14 days.Conclusion:1. The second occipital cistern injection model of autologous caudal vein blood can reproduce the course of SAH.2.Adenoviruses injected into cistern of occipital cistern can improve the CBFs after SAH after down-regulation of Cx43.It can improve the prognosis of SAH.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R651.1

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