Caveolin-1对血管吻合口狭窄的抑制作用及机制
发布时间：2019-06-28 11:59
【摘要】：目的： 建立家兔颈总动脉血管吻合口狭窄模型，并用局部转染小凹蛋白（Caveolin-1）质粒，观察血管吻合口狭窄情况，研究caveolin-1对在血管吻合口狭窄的作用以及与ERK1/2信号通路的关系。方法： 从南华大学动物中心取成年家兔40只，构建血管吻合口再狭窄动物模型，运用脂质体局部转染caveolin-1质粒，随机分为4组：正常组、手术组、空转染组、转染组。于术后第7天取5只兔血管标本，用于westernblot和PCR检测蛋白和mRNA的表达；其余兔于术后4周处死后取标本用于HE染色测量内膜中膜面积比值和免疫组化检测。 结果： 1、组织切片HE染色显示：与正常组相比，手术组内膜明显增殖，管壁增厚，管腔缩小；而与手术组比较，转染组未见明显内膜增殖及管壁增厚，从血管内膜/中膜面积比值发现，转染组比手术组明显降低，说明caveolin-1可抑制血管吻合口狭窄。2、RT-PCR测定caveolin-1和ERK1基因转录，结果显示：与正常组比较,手术组和空转染组的caveolin-1mRNA明显降低（P＜0.01），提示手术使caveolin-1的mRNA表达下调，而手术组ERK1mRNA的表达明显升高，转染组表达明显降低（P＜0.05）3、免疫组化测定caveolin-1表达，结果显示：与正常组比较，手术组和空转染组caveolin-1的表达明显降低，用WesternBlot检测caveolin-1蛋白的表达,与正常组比较，手术组和空转染组的caveolin-1蛋白的表达明显降低（P＜0.05），说明血管损伤可以使caveolin-1表达下调；转染组caveolin-1的表达明显增高，说明caveolin-1转染成功；免疫组化检测磷酸化ERK1/2的表达，，结果显示：与正常组比较，手术组和空转组磷酸化ERK1/2表达明显增高，而转染组明显降低；用蛋白杂交结果进一步证实的同样的趋势。提示caveolin-1可以抑制血管吻合口狭窄其机制可能与抑制ERK1/2活化有关。 结论: 1、Caveolin-1可抑制家兔颈总动脉吻合口术后内膜的增殖。 2、Caveolin-1抑制吻合口狭窄的作用可能与调节ERK的活化有关。
[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
本文编号：2507284
[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
本文编号：2507284
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