【摘要】：目的： 建立家兔頸總動脈血管吻合口狹窄模型，并用局部轉(zhuǎn)染小凹蛋白（Caveolin-1）質(zhì)粒，觀察血管吻合口狹窄情況，研究caveolin-1對在血管吻合口狹窄的作用以及與ERK1/2信號通路的關(guān)系。方法： 從南華大學(xué)動物中心取成年家兔40只，構(gòu)建血管吻合口再狹窄動物模型，運(yùn)用脂質(zhì)體局部轉(zhuǎn)染caveolin-1質(zhì)粒，隨機(jī)分為4組：正常組、手術(shù)組、空轉(zhuǎn)染組、轉(zhuǎn)染組。于術(shù)后第7天取5只兔血管標(biāo)本，用于westernblot和PCR檢測蛋白和mRNA的表達(dá)；其余兔于術(shù)后4周處死后取標(biāo)本用于HE染色測量內(nèi)膜中膜面積比值和免疫組化檢測。 結(jié)果： 1、組織切片HE染色顯示：與正常組相比，手術(shù)組內(nèi)膜明顯增殖，管壁增厚，管腔縮小；而與手術(shù)組比較，轉(zhuǎn)染組未見明顯內(nèi)膜增殖及管壁增厚，從血管內(nèi)膜/中膜面積比值發(fā)現(xiàn)，轉(zhuǎn)染組比手術(shù)組明顯降低，說明caveolin-1可抑制血管吻合口狹窄。2、RT-PCR測定caveolin-1和ERK1基因轉(zhuǎn)錄，結(jié)果顯示：與正常組比較,手術(shù)組和空轉(zhuǎn)染組的caveolin-1mRNA明顯降低（P＜0.01），提示手術(shù)使caveolin-1的mRNA表達(dá)下調(diào)，而手術(shù)組ERK1mRNA的表達(dá)明顯升高，轉(zhuǎn)染組表達(dá)明顯降低（P＜0.05）3、免疫組化測定caveolin-1表達(dá)，結(jié)果顯示：與正常組比較，手術(shù)組和空轉(zhuǎn)染組caveolin-1的表達(dá)明顯降低，用WesternBlot檢測caveolin-1蛋白的表達(dá),與正常組比較，手術(shù)組和空轉(zhuǎn)染組的caveolin-1蛋白的表達(dá)明顯降低（P＜0.05），說明血管損傷可以使caveolin-1表達(dá)下調(diào)；轉(zhuǎn)染組caveolin-1的表達(dá)明顯增高，說明caveolin-1轉(zhuǎn)染成功；免疫組化檢測磷酸化ERK1/2的表達(dá)，，結(jié)果顯示：與正常組比較，手術(shù)組和空轉(zhuǎn)組磷酸化ERK1/2表達(dá)明顯增高，而轉(zhuǎn)染組明顯降低；用蛋白雜交結(jié)果進(jìn)一步證實(shí)的同樣的趨勢。提示caveolin-1可以抑制血管吻合口狹窄其機(jī)制可能與抑制ERK1/2活化有關(guān)。 結(jié)論: 1、Caveolin-1可抑制家兔頸總動脈吻合口術(shù)后內(nèi)膜的增殖。 2、Caveolin-1抑制吻合口狹窄的作用可能與調(diào)節(jié)ERK的活化有關(guān)。
[Abstract]:Aim: to establish a rabbit model of common carotid artery anastomotic stenosis and to observe the effect of caveolin-1 on vascular anastomotic stenosis and its relationship with ERK1/2 signal pathway by local transfection of fovea protein (Caveolin-1) plasmid. Methods: 40 adult rabbits were selected from the Animal Center of Nanhua University to construct the animal model of vascular anastomotic restenosis. Caveolin-1 plasmid was locally transfected with liposomes and randomly divided into four groups: normal group, operation group, empty staining group and transfer group. On the 7th day after operation, the vascular specimens of 5 rabbits were taken to detect the expression of protein and mRNA by westernblot and PCR, and the other rabbits were killed 4 weeks after operation for HE staining to measure the ratio of intima to media area and to detect the ratio of intima to media area. Results: 1. HE staining showed that compared with the normal group, the intima of the operation group proliferated obviously, the wall of the tube thickened and the lumen narrowed. Compared with the operation group, there was no obvious intimal proliferation and wall thickening in the transfer group. It was found that the ratio of intima / media area in the transfer group was significantly lower than that in the operation group, indicating that caveolin-1 could inhibit vascular anastomotic stricture. 2, caveolin-1 and ERK1 gene transcription were measured by RT PCR. The results showed that caveolin-1mRNA in the operation group and empty staining group was significantly lower than that in the normal group (P < 0.01). It is suggested that the expression of mRNA in caveolin-1 was down-regulated by operation, but the expression of ERK1mRNA in operation group was significantly higher than that in transfer group (P < 0.05). The expression of caveolin-1 was detected by immunohistochemistry. The results showed that the expression of caveolin-1 in operation group and empty staining group was significantly lower than that in normal group, and the expression of caveolin-1 protein was detected by WesternBlot compared with normal group. The expression of caveolin-1 protein in operation group and empty staining group was significantly decreased (P < 0.05), which indicated that vascular injury could down-regulate the expression of caveolin-1. The expression of caveolin-1 in the transfer group was significantly higher than that in the control group, indicating that the expression of phosphorylated ERK1/2 in the transfer group was significantly higher than that in the normal group, while the expression of phosphorylated ERK1/2 in the operation group and the empty group was significantly higher than that in the normal group, while that in the transfer group was significantly lower than that in the normal group, and the same trend was further confirmed by the results of protein hybridization. It is suggested that caveolin-1 can inhibit vascular anastomotic stenosis and its mechanism may be related to the inhibition of ERK1/2 activation. Conclusion: 1 Caveolin 鈮

本文編號：2507284