發(fā)布時間：2018-04-10 12:41

  本文選題：創(chuàng)傷性顱腦損傷 + 缺氧誘導因子-1?��； 參考：《天津醫(yī)科大學》2017年博士論文

【摘要】：目的:隨著社會進步與經(jīng)濟的高速發(fā)展及國際局勢的多變,因交通事故、生產(chǎn)意外、自然災害、局部區(qū)域的戰(zhàn)爭等因素造成的創(chuàng)傷性顱腦損傷(Traumatic Brain Injury,TBI)的發(fā)病率越來越高,其致殘率以及致死率逐年上升,特別是由于惡性車禍、生產(chǎn)事故以及殺傷性武器的使用所帶來的中、重度TBI對患者傷害尤為巨大,同時也給家庭以及社會造成很大的傷害與不良影響。因此,對于TBI發(fā)病機制的研究以及對其給予有效的干預、治療方法的探索越來越受到創(chuàng)傷外科、神經(jīng)醫(yī)學等多領(lǐng)域研究學者的關(guān)注。缺氧誘導因子-1?(hypoxia-inducible factor-1?,HIF-1?)與TBI的多種病理變化關(guān)系非常密切。在TBI后缺血缺氧的內(nèi)環(huán)境中,HIF-1?對神經(jīng)細胞的適應、存活、凋亡以及對神經(jīng)血管的修復、再生等病理生理變化過程都起著重要的穩(wěn)定與促進作用�；诖�,本課題擬通過觀察在TBI中HIF-1?是否也發(fā)揮著抑制炎癥反應以及促進血管生成的作用,并通過調(diào)控HIF-1?進一步探討其在TBI中可能存在的神經(jīng)血管保護作用。方法:利用液壓打擊儀建立Wistar大鼠TBI模型,在造模前將實驗動物隨機分為假手術(shù)組、TBI組、HIF-1?沉默表達組以及對照病毒顆粒組。觀察時相分別設(shè)置為模型制備后第3、7、14天(即急性期、亞急性期、慢性期三個時段):(1)通過改良神經(jīng)功能缺損程度評分(mNSS)量表對TBI后的大鼠神志及功能缺損變化進行動態(tài)評估。(2)通過HE切片染色觀察TBI腦損傷面積以及損傷區(qū)形態(tài)結(jié)構(gòu)的變化;(3)通過Western Blot方法檢測大鼠受傷腦組織HIF-1?及vWF的表達變化;(4)通過ELISA法檢測腦組織MMP9、VEGF、TNF-α、IL-6、NF-κB的表達變化;(5)通過統(tǒng)計學觀察HIF-1?對TBI神經(jīng)功能的影響分析其神經(jīng)保護作用,并初步探討HIF-1?與血管生成、炎癥反應相關(guān)的分子機制。結(jié)果:(1)HE染色顯示TBI組腦水腫程度在第3天、第7天、第14天較HIF-1?沉默表達組均有所減輕(P0.05,P0.05,P0.05);而改良神經(jīng)功能缺損評分水平在第7天、第14天TBI組較HIF-1?沉默表達組也均明顯好轉(zhuǎn)(P0.05,P0.05)。(2)TBI大鼠中,HIF-1?沉默慢病毒顆粒顱內(nèi)微量注射預處理后,腦組織內(nèi)HIF-1?與vWF表達水平在3天、7天、14天三個觀察時相均較TBI組明顯下降(P0.05,P0.05,P0.05),表明腦組織內(nèi)HIF-1?沉默表達的成功,且可抑制腦血管生成。液壓打擊造成TBI后第3天,TBI組HIF-1?表達水平較HIF-1?沉默表達組與假手術(shù)組明顯升高(P0.05,P0.05),到第7天與第14天仍然保持較高水平,第7天TBI組HIF-1?表達水平較HIF-1?沉默表達組與假手術(shù)組升高明顯(P0.05,P0.05),第14天TBI組HIF-1?表達水平較HIF-1?沉默表達組與假手術(shù)組也均升高明顯(P0.05,P0.05)。(3)HIF-1?沉默表達組在3天、7天、14天的VEGF活性表達水平與TBI組相比均明顯下降(P0.05,P0.05,P0.05),而MMP-9活性水平在第3天小幅下降(P0.05)后,在第7天、第14天出現(xiàn)顯著的降低(P0.05,P0.05)。(4)IL-6活性水平:TBI組與HIF-1?沉默表達組比較在第3天、第7天、第14天,三個時相點均明顯降低(P0.05,P0.05,P0.05);TNF-α水平:TBI組較HIF-1?沉默表達組在第3天、第7天、第14天,三個時相也明顯降低(P0.05,P0.05,P0.05);NF-κB激活程度在第3天、第7天、第14天,三個時相TBI組較HIF-1?沉默表達組明顯減弱(P0.05,P0.05,P0.05)。結(jié)論:HIF-1?沉默表達組炎癥反應抑制以及促進血管形成方面均較TBI組明顯減弱,從而說明HIF-1?在TBI后可通過抑制炎癥反應以及促進血管形成的雙重調(diào)節(jié)機制降低腦水腫程度,保護腦組織、改善腦神經(jīng)功能,促進TBI恢復。
[Abstract]:Objective: with the rapid development of economic and social progress and the changeable international situation, due to traffic accidents, production accidents, natural disasters, traumatic brain injury caused by local war and other factors (Traumatic Brain, Injury, TBI) the increasing incidence of the disability rate and the mortality rate increased year by year, especially since the malignant accidents, accidents and the use of weapons of destruction caused by severe injury to patients, TBI is particularly great, but also caused great harm and adverse effects to the family and society. Therefore, the study of TBI pathogenesis and effective intervention for the treatment of exploration, more and more trauma surgery researchers in many fields, such as neural medical attention. Hypoxia inducible factor -1 (hypoxia-inducible? Factor-1?, HIF-1?) are associated with a variety of pathological changes. The relationship between TBI in TBI after hypoxia ischemia In the HIF-1 environment,? Adaptation, on cell survival, apoptosis and repair the nerve and blood vessel pathological changes, regeneration process plays an important role in promoting stability and. Based on this, this paper through the observation of HIF-1 in TBI? Whether to inhibit inflammatory reaction and promote angiogenesis the role, and through the regulation of HIF-1? Further discusses the possible TBI in the protection of the nerves and blood vessels. Methods: the use of hydraulic percussion instrument to establish Wistar rat model of TBI, before modeling the experimental animal were randomly divided into sham operation group, TBI group, HIF-1 group and the expression of silence? Virus particles observed group. When the phase were set for the preparation of the model after 3,7,14 days (i.e., acute, subacute, chronic period of three hours): (1) by modified neurological severity scores (mNSS) scale of god records and function defect changes of rats after TBI. Dynamic assessment. (2) to observe the changes of TBI area of brain injury and the morphology damage zone by HE staining; (3) detected by Western Blot method in rats injured brain tissue HIF-1 expression and vWF?; (4) through the detection of brain tissue by ELISA MMP9, VEGF, TNF- alpha, IL-6, expression NF- K B; (5) through statistical observation of the neuroprotective effect of HIF-1? TBI analysis on the influence of neural function, and to explore HIF-1? And angiogenesis, molecular mechanism of inflammatory reaction. Results: (1) HE staining showed that TBI group of brain edema in third days, seventh days, fourteenth days HIF-1? Silent expression group were improved (P0.05, P0.05, P0.05); and the modified neurological score levels in seventh days, fourteenth days in TBI group than HIF-1 group also? Silencing expression were significantly improved (P0.05, P0.05). (2) HIF-1 TBI in rat? Silent lentivirus intracranial microinjection after the pretreatment, brain tissue H IF-1?涓巚WF琛ㄨ揪姘村鉤鍦，

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