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

【摘要】：目的:隨著社會(huì)進(jìn)步與經(jīng)濟(jì)的高速發(fā)展及國(guó)際局勢(shì)的多變,因交通事故、生產(chǎn)意外、自然災(zāi)害、局部區(qū)域的戰(zhàn)爭(zhēng)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(lái)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(lái)的中、重度TBI對(duì)患者傷害尤為巨大,同時(shí)也給家庭以及社會(huì)造成很大的傷害與不良影響。因此,對(duì)于TBI發(fā)病機(jī)制的研究以及對(duì)其給予有效的干預(yù)、治療方法的探索越來(lái)越受到創(chuàng)傷外科、神經(jīng)醫(yī)學(xué)等多領(lǐng)域研究學(xué)者的關(guān)注。缺氧誘導(dǎo)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(duì)神經(jīng)細(xì)胞的適應(yīng)、存活、凋亡以及對(duì)神經(jīng)血管的修復(fù)、再生等病理生理變化過(guò)程都起著重要的穩(wěn)定與促進(jìn)作用�；诖�,本課題擬通過(guò)觀察在TBI中HIF-1?是否也發(fā)揮著抑制炎癥反應(yīng)以及促進(jìn)血管生成的作用,并通過(guò)調(diào)控HIF-1?進(jìn)一步探討其在TBI中可能存在的神經(jīng)血管保護(hù)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(shí)驗(yàn)動(dòng)物隨機(jī)分為假手術(shù)組、TBI組、HIF-1?沉默表達(dá)組以及對(duì)照病毒顆粒組。觀察時(shí)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(gè)時(shí)段):(1)通過(guò)改良神經(jīng)功能缺損程度評(píng)分(mNSS)量表對(duì)TBI后的大鼠神志及功能缺損變化進(jìn)行動(dòng)態(tài)評(píng)估。(2)通過(guò)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(guò)Western Blot方法檢測(cè)大鼠受傷腦組織HIF-1?及vWF的表達(dá)變化;(4)通過(guò)ELISA法檢測(cè)腦組織MMP9、VEGF、TNF-α、IL-6、NF-κB的表達(dá)變化;(5)通過(guò)統(tǒng)計(jì)學(xué)觀察HIF-1?對(duì)TBI神經(jīng)功能的影響分析其神經(jīng)保護(hù)作用,并初步探討HIF-1?與血管生成、炎癥反應(yīng)相關(guān)的分子機(jī)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(dá)組均有所減輕(P0.05,P0.05,P0.05);而改良神經(jīng)功能缺損評(píng)分水平在第7天、第14天TBI組較HIF-1?沉默表達(dá)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(yù)處理后,腦組織內(nèi)HIF-1?與vWF表達(dá)水平在3天、7天、14天三個(gè)觀察時(shí)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(dá)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(dá)水平較HIF-1?沉默表達(dá)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(dá)水平較HIF-1?沉默表達(dá)組與假手術(shù)組升高明顯(P0.05,P0.05),第14天TBI組HIF-1?表達(dá)水平較HIF-1?沉默表達(dá)組與假手術(shù)組也均升高明顯(P0.05,P0.05)。(3)HIF-1?沉默表達(dá)組在3天、7天、14天的VEGF活性表達(dá)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(dá)組比較在第3天、第7天、第14天,三個(gè)時(shí)相點(diǎn)均明顯降低(P0.05,P0.05,P0.05);TNF-α水平:TBI組較HIF-1?沉默表達(dá)組在第3天、第7天、第14天,三個(gè)時(shí)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(gè)時(shí)相TBI組較HIF-1?沉默表達(dá)組明顯減弱(P0.05,P0.05,P0.05)。結(jié)論:HIF-1?沉默表達(dá)組炎癥反應(yīng)抑制以及促進(jìn)血管形成方面均較TBI組明顯減弱,從而說(shuō)明HIF-1?在TBI后可通過(guò)抑制炎癥反應(yīng)以及促進(jìn)血管形成的雙重調(diào)節(jié)機(jī)制降低腦水腫程度,保護(hù)腦組織、改善腦神經(jīng)功能,促進(jìn)TBI恢復(fù)。
[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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