本文關鍵詞： 可卡因-苯丙胺調節(jié)轉錄肽 缺血性腦損傷 氧化應激 線粒體　出處：《南京醫(yī)科大學》2014年碩士論文　論文類型：學位論文

【摘要】：目的：探討可卡因-苯丙胺調節(jié)轉錄肽(cocaine-and amphetamine-regulated transcript peptides, CART)抗氧化減輕缺血性腦損傷及其機制。 方法：健康雄性ICR小鼠隨機分為四組：缺血／再灌注(Ischemia/Reperfusion,I/R)組、CART組、生理鹽水(Normal Saline, NS)組、假手術組。插線法建立大腦中動脈閉塞(middle cerebral artery occlusion, MCAO)模型,CART組和NS組于缺血2小時再灌注前分別經尾靜脈注射給予CART55-102和同體積NS,然后每隔24小時重復給藥1次。再灌注不同時間點分別以TTC染色檢測腦梗死體積,干／濕法測定腦組織含水率,酶聯(lián)免疫吸附法檢測梗死側大腦皮質4-羥壬烯醛(HNE)、8-羥基-2’-脫氧鳥苷(8-OHdG)及3-硝基酪氨酸(3-NT)含量；采用流式細胞技術檢測梗死側大腦皮質氧自由基(ROS)含量和線粒體膜電位；采用比色法檢測線粒體呼吸鏈復合物Ⅰ、Ⅱ、Ⅲ活性；實時聚合酶鏈式反應(RT-PCR)測定梗死側大腦皮質線粒體DNA (mitochondrial DNA, mtDNA) mRNA表達。 結果：(1)與I/R組和NS組比較,CART組再灌注后各時間點腦梗死體積均不同程度減小,再灌注24h、48h、72h其差異均有統(tǒng)計學意義(均P0.05)。(2)與I/R組和NS組比較,CART在再灌注后各時間點(12h、24h、48h、72h)均可降低腦組織含水率(均P0.05)。(3)與I/R組和NS組比較,CART組在各時間點腦組織中的HNE、8-OHdG、3-NT含量均被下調：HNE在各時間點差異均有統(tǒng)計學意義(均P0.01),8-OHdG僅在12h差異有統(tǒng)計學意義(P0.05),3-NT在24h、48h及72h時間點差異有統(tǒng)計學意義(均P0.05)。(4)與I/R組和NS組比較,CART顯著下調缺血腦組織ROS含量(P0.05)。(5)與I/R組和NS組比較,CART顯著上調缺血腦組織線粒體膜電位(P0.01)、線粒體呼吸鏈復合物Ⅱ活性(P0.05)和mtDNA mRNA表達(P0.05)；線粒體呼吸鏈復合物Ⅰ、Ⅲ活性未被上調(P0.05)。 結論：CART抗氧化減輕缺血性腦損傷,其機制可能是通過穩(wěn)定線粒體膜電位、上調線粒體呼吸鏈復合物Ⅱ活性和mtDNA表達發(fā)揮抑制氧化作用。
[Abstract]:Aim: to investigate the antioxidation effect of cocaine-amphetamine-regulated transcription peptide cocaine-and amphetamine-regulated transcript peptides (cart) on ischemic brain injury and its mechanism. Methods: healthy male ICR mice were randomly divided into four groups: ischemic / reperfusion Ischemia / reperfusion I / R group and normal Saline (NSN) group. The middle cerebral artery occlusion (MCAO) model of middle cerebral artery occlusion (MCAO) was established in sham-operation group. Before ischemia 2 hours after reperfusion, CART55-102 and NSN of the same volume were injected into the caudal vein, respectively, in the cart group and NS group, and then repeated every 24 hours. Cerebral infarction volume was measured by TTC staining at different time points of perfusion. The water content of brain tissue was measured by dry / wet method, and the contents of 8-OHdG and 3-nitrotyrosine were detected by enzyme-linked immunosorbent assay (Elisa) in the cerebral cortex of the infarcted side. The contents of 8-OHdG) and 3-nitrotyrosine were detected by enzyme-linked immunosorbent assay. The contents of oxygen free radical and mitochondrial membrane potential in cerebral cortex of infarct side were detected by flow cytometry, and the activities of respiratory chain complex 鈪，

本文編號：1539320