本文關(guān)鍵詞： 缺血再灌注 肢體 脊髓 細(xì)胞凋亡 前列地爾　出處：《山西醫(yī)科大學(xué)》2009年碩士論文　論文類型：學(xué)位論文

【摘要】： 研究背景:肢體缺血再灌注損傷在臨床外科中較多見,研究肢體缺血再灌注損傷具有重要的臨床意義。肢體缺血再灌注除造成肢體自身的損傷外,還通過神經(jīng)-內(nèi)分泌-免疫機(jī)制導(dǎo)致全身炎癥反應(yīng)綜合征,出現(xiàn)遠(yuǎn)隔臟器的功能障礙和損傷,甚至危及生命。肢體缺血再灌注后周圍神經(jīng)發(fā)生一系列結(jié)構(gòu)和功能的異常變化,對(duì)中樞神經(jīng)系統(tǒng)產(chǎn)生重要影響,導(dǎo)致脊髓神經(jīng)元的損傷。 目的:觀察大鼠肢體缺血再灌注脊髓神經(jīng)元損傷過程中細(xì)胞凋亡、Bcl-2和Caspase-3的變化及前列地爾對(duì)此過程的干預(yù)效應(yīng)。 方法:成年健康SD大鼠72只,隨機(jī)分為3組:①假手術(shù)組(sham組,n=24),只進(jìn)行麻醉并解剖股血管和股神經(jīng),不做其他處理。②肢體缺血3h再灌注組(IR組,n=24)。③肢體缺血3h再灌注+前列地爾組( IR+PGE1組,n=24),缺血前10min于大鼠尾靜脈注射前列地爾5μg·ml-1·kg-1。每組按照再灌注時(shí)間(4h、12h、24h、48h)又分為4個(gè)時(shí)相。實(shí)驗(yàn)結(jié)束,采用心臟灌注處死大鼠,取脊髓腰段(L1~5),采用HE染色觀察脊髓病理學(xué)改變;免疫組織化學(xué)法(SP法)觀察Bcl-2和Caspase-3蛋白質(zhì)表達(dá)情況; TUNEL法觀察細(xì)胞凋亡的發(fā)生情況。 結(jié)果:①脊髓組織中Bcl-2和Caspase-3的表達(dá):免疫組織化學(xué)切片以陽性細(xì)胞率為指標(biāo)進(jìn)行定量分析。sham組Bcl-2和Caspase-3表達(dá)均較低。Bcl-2表達(dá)陽性細(xì)胞率定量分析:其它組與sham組比較,Bcl-2陽性細(xì)胞率明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.01);與IR組比較,IR+PGE1組陽性細(xì)胞率增加,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。IR、IR+PGE1組Bcl-2表達(dá)均于12h達(dá)高峰,之后下降。Caspase-3表達(dá)陽性細(xì)胞率定量分析:其它組與sham組比較Caspase-3陽性細(xì)胞率均明顯增加,差異有統(tǒng)計(jì)學(xué)意義(P0.01);與IR組比較,IR+PGE1組陽性細(xì)胞率降低,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。IR、IR+PGE1組Caspase-3表達(dá)于24h達(dá)高峰,之后下降。②TUNEL法檢測(cè)凋亡細(xì)胞:sham組較少見凋亡細(xì)胞。其余各組凋亡指數(shù)(AI)與sham組比較差異均具有統(tǒng)計(jì)學(xué)意義(P0.01);與IR組比較,IR+PGE1組各時(shí)相AI值明顯減少,差別有顯著性,具有統(tǒng)計(jì)學(xué)意義(P0.05)。相關(guān)性分析Caspase-3陽性細(xì)胞率與AI值成正相關(guān)。 結(jié)論:①大鼠肢體缺血再灌注后可導(dǎo)致脊髓神經(jīng)元細(xì)胞凋亡的發(fā)生,引起凋亡相關(guān)蛋白Bcl-2和Caspase-3的表達(dá)增加。②缺血前預(yù)防性給予前列地爾對(duì)大鼠肢體缺血再灌注引起的脊髓神經(jīng)元細(xì)胞凋亡有一定的抑制作用,可能通過上調(diào)Bcl-2的表達(dá),下調(diào)Caspase-3的表達(dá),從而減輕脊髓神經(jīng)元的損傷,起到保護(hù)作用。
[Abstract]:Background: limb ischemia-reperfusion injury is more common in clinical surgery. It has important clinical significance to study limb ischemia-reperfusion injury. It also leads to systemic inflammatory response syndrome through neuroendocrine and immunological mechanism, resulting in dysfunction and injury of distant organs, and even endangering life. A series of abnormal changes in structure and function of peripheral nerves occur after limb ischemia and reperfusion. Has an important effect on the central nervous system, resulting in spinal cord neuron injury. Aim: to observe the changes of apoptosis Bcl-2 and Caspase-3 during spinal cord neuron injury after limb ischemia reperfusion in rats and the effect of alprostadil on this process. Methods: Seventy-two adult healthy SD rats were randomly divided into 3 groups: sham group (n = 24), sham group (n = 3), anesthetized and dissected femoral vessels and femoral nerves. 2. No other treatment. 2 limb ischemia / reperfusion group / IR group / IR / PGE1 group after 3 h / 3 limb ischemia / reperfusion. Ten minutes before ischemia, alprostadil 5 渭 g 路ml-1 路kg ~ (-1) was injected into the tail vein of rats. Each group was divided into 4 groups according to the reperfusion time of 4 h, 12 h, 24 h and 48 h). At the end of the experiment, The rats were killed by cardiac perfusion. The pathological changes of spinal cord were observed by HE staining. The expression of Bcl-2 and Caspase-3 protein was observed by immunohistochemical method (SP method) and apoptosis was observed by TUNEL method. Results the expression of Bcl-2 and Caspase-3 in the spinal cord tissue of 1: 1: immunohistochemical section was used as an index to quantitatively analyze the positive cell rate. The expression of Bcl-2 and Caspase-3 in Sham group was lower than that in the sham group. The positive cell rate of Bcl-2 expression in the other groups was compared with that in the sham group. The rate of Bcl-2 positive cells increased significantly. Compared with IR group, the positive cell rate of IR PGE1 group increased, and the Bcl-2 expression of IR PGE1 group reached its peak at 12 h. After that, the positive cell rate of Caspase-3 decreased: compared with sham group, the positive cell rate of Caspase-3 increased significantly in other groups, the difference was statistically significant (P 0.01), and the positive cell rate in IR PGE1 group was lower than that in IR group. The expression of Caspase-3 in IR PGE1 group reached its peak at 24 h. The apoptosis index of other groups was significantly higher than that of sham group (P 0.01), and the AI value of IR PGE1 group was significantly lower than that of IR PGE1 group (P < 0.05). The correlation analysis showed that the positive cell rate of Caspase-3 was positively correlated with AI value. Conclusion the apoptosis of spinal cord neurons can be induced by ischemia and reperfusion of rat limbs. The expression of apoptosis-related proteins (Bcl-2 and Caspase-3) was increased by prophylaxis of prostadil before ischemia. 2 the apoptosis of spinal cord neurons induced by ischemia and reperfusion of rat limbs was inhibited to some extent, possibly by upregulating the expression of Bcl-2. The expression of Caspase-3 was down-regulated, which alleviated the injury of spinal cord neurons and played a protective role.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R363

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 康一凡，高建章，吳岳嵩，趙定麟，，崔海平;肢體缺血再灌注損傷局部與主要器官脂質(zhì)過氧化的觀察[J];第二軍醫(yī)大學(xué)學(xué)報(bào);1996年05期

2 黃優(yōu)青;顧洪生;;聯(lián)合用藥對(duì)實(shí)驗(yàn)性肢體缺血再灌注損傷復(fù)流后神經(jīng)及肌肉功能恢復(fù)的影響[J];廣東醫(yī)學(xué);1996年05期

3 顏學(xué)軍;細(xì)胞凋亡的線粒體機(jī)制[J];國外醫(yī)學(xué).麻醉學(xué)與復(fù)蘇分冊(cè);2001年03期

4 應(yīng)雋,王祥瑞;補(bǔ)體激活在組織缺血/再灌注損傷中的作用[J];國外醫(yī)學(xué).麻醉學(xué)與復(fù)蘇分冊(cè);2004年03期

5 王彥春,王敏,段妍君,馬駿,王華;大鼠肢體IRI后脊髓神經(jīng)元HSP_(70)的表達(dá)及丹參注射液干預(yù)后的影響[J];湖北中醫(yī)雜志;2004年04期

6 馮國清;劉潔;傅潤芳;翁世艾;;前列腺素E_1對(duì)大鼠心肌缺血再灌注損傷的保護(hù)作用[J];河南醫(yī)學(xué)研究;1997年03期

7 張靈芝,湯健,唐朝樞,蘇靜怡;均勻單層脂質(zhì)體的制備方法[J];基礎(chǔ)醫(yī)學(xué)與臨床;1992年04期

8 陳建常,史振滿,王樂農(nóng),盧開柏;肢體缺血再灌注致遠(yuǎn)處器官損傷及丹參、甘露醇的保護(hù)作用[J];臨床軍醫(yī)雜志;2001年02期

9 李小冰,劉岳文,郅興義;Caspase家族與心臟移植細(xì)胞凋亡[J];河南科技大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2004年01期

10 劉愛芬,馬平;細(xì)胞凋亡與缺血再灌注損傷[J];山西醫(yī)藥雜志;2005年09期

本文編號(hào)：1539557