Caveolin-1对血管吻合后TNF-a、IL-6、IL-10表达的影响

发布时间：2018-05-15 05:04

本文选题：Caveolin-1 + 肿瘤坏死因子-α　；参考：《南华大学》2012年硕士论文

【摘要】：目的探讨Caveolin-1对血管吻合术后TNF-α、IL-6和IL-10表达的影响。方法本实验对家兔颈总动脉离断后行端端吻合手术，建立血管吻合的动物模型。40只成年家兔取自南华大学实验动物部，随机分为4组：正常组、手术组、空转染组、Caveolin-1转染组。分别于术后7d、14d处死家兔，取术后7d标本用于实时荧光定量PCR和Western Blot检测，14d的标本用于HE染色及免疫组化。采用苏木精-伊红染色，观察吻合口血管动脉内膜变化情况；分别应用实时荧光定量PCR、Western Blot和免疫组化检测家兔吻合术后血管组织中Caveolin-1、TNF-α、IL-6、IL-10的表达水平。结果1、组织切片HE染色形态学观察，手术组和空转染组术后14d血管内膜增生明显，血管壁增厚；而转染组血管内膜仅有局部增生，血管壁增厚不如手术组和空转染组明显。2、实时荧光定量PCR显示，Caveolin-1转染组TNF-α、IL-6mRNA表达水平明显低于手术组和空转染组,但高于对照组（P＜0.05）。Caveolin-1转染组IL-10mRNA表达水平明显高于手术组、空转染组和对照组（P＜0.05）。与正常组比较，手术组和空转染组中Caveolin-1mRNA表达明显降低(P0.05)，而转染组Caveolin-1mRNA表达明显高于手术组(P0.05)。3、WesternBlot检测显示Caveolin-1在各组均有表达，与正常组比较，手术组和空转染组中Caveolin-1蛋白表达明显降低(P0.05)，而转染组Caveolin-1蛋白表达明显高于手术组(P0.05)。与正常组比较，手术组和空转染组中TNF-α、IL-6蛋白表达明显增强(P0.05)，转染组TNF-α、IL-6蛋白表达较手术组明显降低(P0.05)。与正常组比较，手术组和空转染组中IL-10蛋白表达增强(P0.05)，转染组IL-10蛋白表达较手术组明显增强(P0.05)。4、免疫组化染色显示转染组TNF-α、IL-6表达较手术组和空转染组明显降低，，IL-10的表达较手术组和空转染组明显增强。结论1、Caveolin-1能抑制家兔颈总动脉吻合口术后内膜的增殖。 2、Caveolin-1可以抑制血管吻合口术后血管组织的TNF-α、IL-6的表达，同时上调IL-10的表达，调节炎症反应。
[Abstract]:Objective to investigate the effect of Caveolin-1 on the expression of TNF-, IL-6 and IL-10 after vascular anastomosis.
Methods the animal model of vascular anastomosis was established after the end end anastomosis of the common carotid artery in the rabbit. The animal model.40 was taken from the experimental animal Department of University of South China and divided into 4 groups randomly: the normal group, the operation group, the empty transfection group and the Caveolin-1 transfection group. After the operation, the rabbits were killed by 7d and 14d, and the 7d specimens were used for real-time fluorescence quantitative analysis. PCR and Western Blot were detected, and the specimens of 14d were used for HE staining and immunohistochemistry. The changes of vascular intima were observed by hematoxylin eosin staining, and the expression of Caveolin-1, TNF- a, IL-6, IL-10 was detected by real-time fluorescent quantitative PCR, Western Blot and immunohistochemistry.
Results 1, the morphological observation of tissue section HE staining showed that the vascular intima hyperplasia was obvious and the vascular wall thickened after the operation group and the empty transfection group, but the vascular intima was only localized in the transfected group, and the thickening of the vascular wall was not as obvious as that of the operation group and the empty transfected group. The real-time fluorescence quantitative PCR showed that the Caveolin-1 transfected group was TNF- alpha and the expression of IL-6mRNA was obvious. The expression of IL-6mRNA was obvious in the Caveolin-1 transfected group. The expression level of IL-10mRNA in the transfected group was significantly higher than that in the control group (P < 0.05).Caveolin-1 transfection group. The expression of Caveolin-1mRNA in the operation group and the empty transfected group was significantly lower than that in the control group (P < 0.05). The expression of Caveolin-1mRNA in the transfected group was significantly lower than that in the normal group (P0.05), but the expression of Caveolin-1mRNA in the transfected group was significantly higher than that of the operation group (P0). .05).3, WesternBlot detection showed that Caveolin-1 was expressed in all groups. Compared with the normal group, the expression of Caveolin-1 protein in the operation group and the empty transfected group was significantly decreased (P0.05), while the expression of Caveolin-1 protein in the transfected group was significantly higher than that of the operation group (P0.05). The expression of TNF- alpha and IL-6 protein in the hand and air transfected groups was obviously enhanced (P0.0). 5), the expression of TNF- alpha and IL-6 protein in the transfected group was significantly lower than that in the operation group (P0.05). Compared with the normal group, the expression of IL-10 protein in the operation group and the empty transfected group was enhanced (P0.05). The expression of IL-10 protein in the transfected group was significantly enhanced (P0.05).4, and the immunohistochemical staining showed TNF- alpha in the transfer group, and the expression of IL-6 was significantly lower than that in the operation group and the empty transfected group. IL-10 The expression was significantly higher than that in the operation group and the empty transfected group.
Conclusion 1, Caveolin-1 can inhibit intimal proliferation after carotid artery anastomosis in rabbits.
2, Caveolin-1 can inhibit the expression of TNF- and IL-6 in vascular tissue after vascular anastomosis, and increase the expression of IL-10 and regulate the inflammatory response.

【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R363;R-332

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