本文選題：法醫(yī)病理學(xué)　切入點(diǎn)：DAI　出處：《華中科技大學(xué)》2013年碩士論文　論文類型：學(xué)位論文

【摘要】：【研究背景、目的】 彌漫性軸索損傷（diffuse axonal injury， DAI），也稱“彌漫性軸突損傷”，是外力引起的廣泛性腦白質(zhì)軸索損傷。腦外傷死亡病人中，DAI占29％～43％。DAI以頭部外傷后意識(shí)障礙為其典型表現(xiàn)，無腦挫裂傷等常見的局灶性損傷癥狀和體征，CT、MRI等常見影像學(xué)檢查不能直接顯示軸索損傷，目前DAI尚無統(tǒng)一的診斷標(biāo)準(zhǔn)。雖然近20年來對(duì)顱腦損傷研究有了很大進(jìn)展，診斷技術(shù)及治療措施也在不斷完善，但顱腦損傷的病死率及致殘率仍未見明顯下降。大量的研究顯示DAI與Ca~(2+)超載、軸漿運(yùn)輸障礙、線粒體損傷、軸索的細(xì)胞骨架破壞、自由基、炎癥因子等有關(guān)。其中Ca~(2+)超載被認(rèn)為是導(dǎo)致軸索損傷瀑布反應(yīng)過程中的中樞環(huán)節(jié)。在治療上，DAI迄今尚無特異方法能夠阻斷DAI進(jìn)展，因此預(yù)后較差。DAI臨床及基礎(chǔ)研究的進(jìn)展緩慢也限制了其在法醫(yī)鑒定中的應(yīng)用，實(shí)際檢案中DAI應(yīng)用較少，目前仍然是法醫(yī)學(xué)鑒定的難點(diǎn)及爭(zhēng)議的焦點(diǎn)。 因此，本研本實(shí)驗(yàn)擬通過在動(dòng)物模型中使用Ca~(2+)拮抗劑尼莫地平（Nimodepine,ND）阻斷來觀察其對(duì)軸索和血管損傷的影響來探討Ca~(2+)在DAI發(fā)病中確切作用及機(jī)制，以期為DAI臨床診治及法醫(yī)學(xué)應(yīng)用提供研究基礎(chǔ)。 【材料和方法】 將28只雄性SD實(shí)驗(yàn)大鼠隨機(jī)分為對(duì)照組、實(shí)驗(yàn)組及干預(yù)組。參照Marmarou實(shí)驗(yàn)動(dòng)物模型，建立大鼠DAI模型，實(shí)驗(yàn)組及干預(yù)組打擊后分別在12h、24h及72h處死，對(duì)照組為假手術(shù)組（大鼠未進(jìn)行打擊），實(shí)驗(yàn)組及干預(yù)組為直線加速度打擊負(fù)荷組，實(shí)驗(yàn)組打擊后不作處理，干預(yù)組打擊并加用尼莫地平（Nimodipine，ND）進(jìn)行干預(yù)。每組每個(gè)時(shí)間點(diǎn)4只。用過量10%水合氯醛腹腔麻醉處死，，開顱取腦，打開顱腔取出大腦、小腦及腦干，沿正中矢狀面將大腦、腦干及小腦分為左、右兩半，其中一半腦用于腦濕干比例測(cè)定，另外一半腦用于組織形態(tài)學(xué)及免疫組化觀察：（1）取一部分腦組織稱重后放入烤箱內(nèi)（85℃）烘烤24小時(shí)，在烘干后再次進(jìn)行腦稱重，進(jìn)行腦濕干比例計(jì)算；（2）另一半腦組織用4%多聚甲醛固定24h，然后進(jìn)行脫水、透明、包埋等制成石蠟切片，用蘇木素-伊紅（HE）染色、β-APP及vWF免疫組織化學(xué)染色進(jìn)行組織學(xué)及免疫組化觀察。 【結(jié)果】實(shí)驗(yàn)組及干預(yù)組腦組織濕干比例呈先升高后降低的趨勢(shì)，腦組織含水量最高值出現(xiàn)在打擊后24h,此后腦組織含水量呈下降趨勢(shì)；給予Nimodepine（ND）干預(yù)后，腦水腫成下降趨勢(shì)，但仍然高于對(duì)照組：實(shí)驗(yàn)組與干預(yù)組HE染色見軸索變粗、扭曲、腫脹、斷裂且隨時(shí)間呈逐漸加重趨勢(shì)，干預(yù)組給予ND干預(yù)后軸索損傷程度有所減輕，但仍高于對(duì)照組；實(shí)驗(yàn)組與干預(yù)組各時(shí)間點(diǎn)β-APP免疫組織化學(xué)染色見β-APP表達(dá)隨時(shí)間延長(zhǎng)表達(dá)增強(qiáng)，干預(yù)組給予ND干預(yù)后β-APP表達(dá)有所減少，但仍高于對(duì)照組；vWF表達(dá)部位位于蛛網(wǎng)膜下腔、腦實(shí)質(zhì)血管內(nèi)皮細(xì)胞及腦干部位軸索，打擊后vWF表達(dá)明顯增強(qiáng)，24h尤其明顯，腦實(shí)質(zhì)血管內(nèi)皮細(xì)胞陽性表達(dá)較強(qiáng)的區(qū)域與腦水腫嚴(yán)重區(qū)域較一致，尼莫地平干預(yù)減輕vWF表達(dá)，特別是腦實(shí)質(zhì)血管的表達(dá)顯著減輕。 【結(jié)論】本研究模擬Marmarou動(dòng)物模型構(gòu)建DAI模型，觀察打擊后出現(xiàn)腦水腫、軸索損傷并見點(diǎn)灶狀蛛網(wǎng)膜下腔出血；運(yùn)用鈣離子拮抗劑ND干預(yù)后可以減輕腦水腫和軸索損傷并血管損傷局限化，提示鈣離子信號(hào)在彌漫性軸索損傷中有重要作用；腦水腫在打擊后24h達(dá)到高峰，推測(cè)創(chuàng)傷后24h可能是治療彌漫性軸索損傷的較佳時(shí)間窗。
[Abstract]:[research background, purpose]
Diffuse axonal injury (diffuse axonal, injury, DAI), also known as "diffuse axonal injury", is the external cause of extensive cerebral white matter axonal injury. The patient died of cerebral trauma, DAI accounted for 29% ~ 43%.DAI in head injury of disturbance of consciousness is the typical performance, no brain contusion and other common office focal injury symptoms and signs, CT, MRI and other common imaging cannot directly display axonal injury, at present there is no uniform diagnostic criteria of DAI. Although nearly 20 years study of brain injury has made great progress, diagnostic techniques and treatment measures are continuously improved, but the mortality rate of brain injury and the disability rate is there was no obvious decline. A large number of studies show that DAI and Ca~ (2+) overload, damage to mitochondria impaired axonal transport, cytoskeleton, cable shaft damage, free radicals, inflammatory factors and so on. The Ca~ (2+) overload is considered to be the cause of axonal injury cascade The central link in the process. In the treatment of DAI, so far there is no specific method to block the progression of DAI, so.DAI has a poor prognosis in clinical and basic research of the slow also limits its application in forensic identification, the DAI application is less practical cases, is still the focus of forensic identification of the difficulties and disputes.
Therefore, the research in this study through the use of Ca~ in animal models (2+) antagonist nimodipine (Nimodepine, ND) to observe the effect of blocking axonal and vascular injury of Ca~ (2+) and the exact mechanism in the pathogenesis of DAI, in order to provide the basis for the study of DAI clinical diagnosis and forensic medicine the application of science.
[materials and methods]
28 male SD rats were randomly divided into control group, experimental group and intervention group. According to the Marmarou experimental animal model DAI rat model, the experimental group and the intervention group after the attack at 12h, 24h and 72h were control group (sham operation group rats were not hit), experiment group and intervention group for linear acceleration combat load group, the experimental group against no treatment, the intervention group combat and added nimodipine (Nimodipine, ND) to intervene. In each group 4 rats at each time point. With an excess of 10% chloral hydrate intraperitoneal anesthesia were craniotomy for brain, open the cranial cavity out of the brain, cerebellum and brainstem the brain, along the sagittal plane, brainstem and cerebellum is divided into left and right halves, one half of the brain to brain wet to dry ratio determination, the other half of the brain for histological and immunohistochemical observation: (1) take part in brain tissue after weighing into a baking box (85 DEG C) baking 24 hours in baking Dry again after brain weight, brain wet dry ratio; (2) the other half of the brain tissue was fixed with 4% paraformaldehyde 24h, then dehydrated, transparent, etc. embedded into paraffin sections, with hematoxylin eosin (HE) staining, beta -APP and vWF immunohistochemical staining of tissue pathological and immunohistochemical observation.
[results] the experimental group and the intervention group of brain tissue wet to dry ratio first increased and then decreased, the water content of brain tissue had the highest value in the fight against 24h, then the brain tissue water content decreased; Nimodepine (ND) intervention, brain edema and a downward trend, but still higher than the control group: the experiment group and intervention group HE staining showed axonal diameter, distortion, swelling, fracture and showed a gradually increasing trend in the intervention group were given ND intervention to axonal damage has been reduced, but still higher than the control group; the experimental group and the intervention group at different time points P -APP immunohistochemical staining showed the expression of -APP beta with time prolonged expression, the intervention group was given ND after the intervention of beta -APP expression decreased, but still higher than the control group; the expression of vWF located in the subarachnoid space, cerebral vascular endothelial cells and brain stem axons, the expression of vWF was significantly enhanced after the attack, especially 24h Obviously, the areas with strong positive expression of cerebral parenchymal vascular endothelial cells were more consistent with severe areas of cerebral edema. Nimodipine intervention alleviated the expression of vWF, especially the expression of cerebral parenchymal vessels.
[Conclusion] model to construct the DAI model this study simulated Marmarou animal brain edema were observed after the attack, axonal injury and focal subarachnoid hemorrhage; use of calcium antagonists after ND intervention can alleviate brain edema and axonal injury and localized vascular injury, suggesting that calcium signaling in diffuse axonal there is an important role in the fight against injury; brain edema after 24h and reached the peak at 24h after trauma that may be a better treatment of diffuse axonal injury time window.

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類號(hào)】：D919.4

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 劉洪恩,孫曉川;β淀粉樣前體蛋白與彌漫性軸索損傷[J];創(chuàng)傷外科雜志;2005年01期

2 朱金龍,朱少華,任亮,劉良,周亦武,陳虎,鄧偉年;大鼠彌散性軸索損傷后β-APP的表達(dá)[J];法醫(yī)學(xué)雜志;2005年03期

3 王志利;侯林生;;腦彌漫性軸索損傷的研究現(xiàn)狀及新進(jìn)展[J];海南醫(yī)學(xué);2007年07期

4 馬付堅(jiān);急性腦梗死vWF、P-選擇素的作用研究[J];腦與神經(jīng)疾病雜志;2002年03期

5 趙新亮;王毅;馮潔;張建寧;;尼莫地平對(duì)彌漫性軸索損傷大鼠腦水腫的影響[J];天津醫(yī)藥;2012年07期

6 黎會(huì);譚源福;;彌漫性軸索損傷動(dòng)物模型的現(xiàn)狀及研究進(jìn)展[J];微創(chuàng)醫(yī)學(xué);2010年01期

7 王鵬;謝榮厚;張賽;;腦彌散性軸索損傷腦水腫研究進(jìn)展[J];武警醫(yī)學(xué)院學(xué)報(bào);2008年08期

8 范學(xué)文,孔凡元,潘月英,劉菊年;急性腦梗死患者腦脊液及血漿中GMP-140、vWF含量的研究[J];寧夏醫(yī)學(xué)院學(xué)報(bào);1999年04期

9 王洪財(cái);馬延斌;;彌漫性軸索損傷模型研究現(xiàn)狀及進(jìn)展[J];中國(guó)臨床神經(jīng)外科雜志;2009年01期

10 洪欣,李鳳芝,尹昭云,顏培華,何一心;不同應(yīng)激損傷所致血管內(nèi)皮細(xì)胞中vWF的變化[J];中國(guó)應(yīng)用生理學(xué)雜志;2000年04期

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