本文關(guān)鍵詞：緊密連接蛋白，，由筆耕文化傳播整理發(fā)布。

摘要：

目的 探討腦出血后血腦屏障微血管內(nèi)皮細(xì)胞間緊密連接蛋白o(hù)ccludin的表達(dá)變化. 方法 將SD雄性大鼠按隨機(jī)數(shù)字表法分為正常對(duì)照組和腦出血組,再按時(shí)間因素將腦出血組分為出血后6h、24h、48h、72h、7d、14d6個(gè)亞組.采用腦內(nèi)注入自體血法制作腦出血模型.HE染色觀察腦出血后血腫周圍腦組織的形態(tài)學(xué)改變；透射電鏡觀察腦出血后血腫周圍緊密連接的超微結(jié)構(gòu)改變；免疫熒光染色檢測(cè)腦出血后血腫周圍緊密連接蛋白o(hù)ccludin的表達(dá)分布狀況；定量RT-PCR檢測(cè)腦出血后血腫周圍腦組織中occludin mRNA的表達(dá)狀況. 結(jié)果 與正常對(duì)照組相比,腦出血組血腫周圍腦組織出現(xiàn)水腫,在48 h左右尤為明顯,局部可見明顯腦細(xì)胞壞死及炎細(xì)胞浸潤(rùn).腦出血后血腫周圍緊密連接發(fā)生明顯破壞,內(nèi)皮細(xì)胞間出現(xiàn)裂隙.免疫熒光染色結(jié)果顯示:正常對(duì)照組緊密連接蛋白o(hù)ccludin呈強(qiáng)陽(yáng)性表達(dá).腦出血后6hoccludin的表達(dá)即開始下降,呈陽(yáng)性表達(dá);腦出血后24～72h occludin的表達(dá)維持在較低水平,呈弱陽(yáng)性表達(dá).定量RT-PCR結(jié)果顯示:腦出血后血腫周圍腦組織中occludin mRNA相對(duì)含量明顯降低,在6～72 h持續(xù)維持在較低水平,與正常對(duì)照組比較差異均有統(tǒng)計(jì)學(xué)意義(P＜0.05). 結(jié)論 腦出血發(fā)生后,緊密連接蛋白occludin 的表達(dá)下降,這可能是腦出血發(fā)生后血腦屏障破壞及腦水腫發(fā)生發(fā)展的重要分子基礎(chǔ)之一.

Abstract： 

Objective To investigate the changes of tight junction (TJ) protein occludin expression between the mierovascular endothelial cells of the blood-brain barrier (BBB) of rats after intracerebral hemorrhage (ICH). Methods Ninety-three male SD rats were randomly divided into normal control group and experimental group; rats of the experimental group were divided into 6 subgroups (6, 24, 48 and 72 h, and 7 and 14 d after intraeerebral hemorrhage). Rat models of intracerebral hemorrhage in the experimental group were established by autologous blood injection.Morphology of the brain tissues was detected by hematoxylin-eosin (HE) staining.The tight junctions between the microvascular endothelial cells of the BBB were sampled for ultrastructural observation using electron microcopy.Immunofluorescence and quantitative real time-PCR were used to analyze the protein and mRNA expressions of occludin,respectively. Results The brain edema was found in brain tissues around the hematoma. Necrosis and inflammatory infiltration could be found in perihematomal brain tissues and it was most obvious at 48 h after intracerebral hemorrhage. Obvious paracellular clefts were found between the adjacent endothelial cells in experimental groups. Strong expression of occludin was noted in the normal brain tissue; expression of occludin was positive 6 h after the injection and weak positive at 24, 48 and 72 h after the injection in the experimental groups.Quantitative real time-PCR showed that the mRNA expression of occludin in the experimental group was significantly reduced as compared with the normal brain tissue at 6,24,48 and 72 h after the injection with statistically significant difference (P＜0.05). Conclusion After intra-cerebral hemorrhage,as a major component of TJ of the BBB,occludin expression is down-regulated,which may be one of the most impotant molecular basis of disruption of BBB.

  本文關(guān)鍵詞：緊密連接蛋白，由筆耕文化傳播整理發(fā)布。