本文選題：Caveolin-1 + 腫瘤壞死因子-α��； 參考：《南華大學》2012年碩士論文

【摘要】：目的探討Caveolin-1對血管吻合術(shù)后TNF-α、IL-6和IL-10表達的影響。 方法本實驗對家兔頸總動脈離斷后行端端吻合手術(shù)，建立血管吻合的動物模型。40只成年家兔取自南華大學實驗動物部，隨機分為4組：正常組、手術(shù)組、空轉(zhuǎn)染組、Caveolin-1轉(zhuǎn)染組。分別于術(shù)后7d、14d處死家兔，取術(shù)后7d標本用于實時熒光定量PCR和Western Blot檢測，14d的標本用于HE染色及免疫組化。采用蘇木精-伊紅染色，觀察吻合口血管動脈內(nèi)膜變化情況；分別應用實時熒光定量PCR、Western Blot和免疫組化檢測家兔吻合術(shù)后血管組織中Caveolin-1、TNF-α、IL-6、IL-10的表達水平。 結(jié)果1、組織切片HE染色形態(tài)學觀察，手術(shù)組和空轉(zhuǎn)染組術(shù)后14d血管內(nèi)膜增生明顯，血管壁增厚；而轉(zhuǎn)染組血管內(nèi)膜僅有局部增生，血管壁增厚不如手術(shù)組和空轉(zhuǎn)染組明顯。2、實時熒光定量PCR顯示，Caveolin-1轉(zhuǎn)染組TNF-α、IL-6mRNA表達水平明顯低于手術(shù)組和空轉(zhuǎn)染組,但高于對照組（P＜0.05）。Caveolin-1轉(zhuǎn)染組IL-10mRNA表達水平明顯高于手術(shù)組、空轉(zhuǎn)染組和對照組（P＜0.05）。與正常組比較，手術(shù)組和空轉(zhuǎn)染組中Caveolin-1mRNA表達明顯降低(P0.05)，而轉(zhuǎn)染組Caveolin-1mRNA表達明顯高于手術(shù)組(P0.05)。3、WesternBlot檢測顯示Caveolin-1在各組均有表達，與正常組比較，手術(shù)組和空轉(zhuǎn)染組中Caveolin-1蛋白表達明顯降低(P0.05)，而轉(zhuǎn)染組Caveolin-1蛋白表達明顯高于手術(shù)組(P0.05)。與正常組比較，手術(shù)組和空轉(zhuǎn)染組中TNF-α、IL-6蛋白表達明顯增強(P0.05)，轉(zhuǎn)染組TNF-α、IL-6蛋白表達較手術(shù)組明顯降低(P0.05)。與正常組比較，手術(shù)組和空轉(zhuǎn)染組中IL-10蛋白表達增強(P0.05)，轉(zhuǎn)染組IL-10蛋白表達較手術(shù)組明顯增強(P0.05)。4、免疫組化染色顯示轉(zhuǎn)染組TNF-α、IL-6表達較手術(shù)組和空轉(zhuǎn)染組明顯降低，，IL-10的表達較手術(shù)組和空轉(zhuǎn)染組明顯增強。 結(jié)論1、Caveolin-1能抑制家兔頸總動脈吻合口術(shù)后內(nèi)膜的增殖。 2、Caveolin-1可以抑制血管吻合口術(shù)后血管組織的TNF-α、IL-6的表達，同時上調(diào)IL-10的表達，調(diào)節(jié)炎癥反應。
[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.

【學位授予單位】：南華大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R363;R-332

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