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研究生:王富文
研究生(外文):Wang, Fu-Wen
論文名稱:餵食高膽固醇的兔子其動脈硬化形成的相關基因表現
論文名稱(外文):Gene expression associated with atherogenesis in high cholesterol-fed rabbits
指導教授:陳玉怜江美治江美治引用關係
指導教授(外文):Chen, Yuh-LienJiang, Meei-Jyh
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:解剖學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:1994
畢業學年度:82
語文別:中文
中文關鍵詞:高膽固醇動脈硬化
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在動脈硬化的血管壁上主要有三種特徵為脂肪堆積、泡沫細胞形成及平滑肌細胞的增生。近來的研究報告顯示immediate-early(IE)gene及extracellular matrix protein(EMP) gene 的基因表現與血管壁的組成細胞之附著、遷移及增生有密切的關係,而且也可能動脈硬化的進展有關。其中c-fos和 fibronectin分別是IE和EMP中的一種。
本實驗的目的是觀察C-J0S和 fibronectin的基因表現與動脈硬化形成的相關性。我們使用雄性紐西蘭白兔為實驗動物,在飼料中添加2%膽固醇及6%花生油以誘導動脈硬化形成,當為實驗組。同時,以餵養一般飼料或在飼料中添加6%花生油為對照組I及II。隨後餵養一週、三週或六週後,將動物犧牲,取出主動脈弓、胸主動脈、腹主動脈與肺動脈幹。分離所得的RNA,以北方墨漬法及定位雜合實驗以研究C-foS和 fibronectin mRNA的表現強度及在血管壁上的分布情形。
實驗組的血清膽固醇與三酸甘油酯的濃度較對照組有顯著增加。同時,在六週的實驗組的血管壁上出現明顯的動脈硬化的病變,其中尤以主動脈弓和肺動脈幹為最顯著。
以北方墨漬法分析在餵食六個星期的高膽固醇飼料的實驗組,其fibronectin mRNA的基因表現,在各血管均比對照組I增加許多;在主動脈弓及肺動脈幹亦較對照組II為多。c-fos mRNA的表現,在一星期及三星期的餵養階段,三組並無明顯差異,但在六星期餵養後,主脈各部位之c-fos mRNA之表現強度,還低於對照組II。
定位雜合實驗顯示主動脈弓的fibronectin mRNA表現量在餵養一星期及三星期後,各組間的差異不大。在餵食高膽固醇飼料六星期的實驗組其血管內膜加厚,fibronectin的基因表現量明顯增加,尤以管內膜的泡沫細胞分布較多。c-fos mRNA在餵養一星期的實驗組其肺動脈幹,幾乎沒有表現。在餵養三週的實驗組呈均勻分布,而在餵食六星期的實驗組,其血管內膜增厚,在三層的血管壁上有顯著均勻分布。fibronectin和c-fos的基因表現在內皮細胞、泡沫細胞、平滑肌細胞和纖維母細胞均有分布。以上的結果顯示fibronectin和c-fos基因表現在動脈硬化過程中有明顯變化,其活化可能與動脈硬化粥瘤的發展有關。
The atherosclerotic lesions are characterized by the accumulation of the lipid, the deposition of the foam cells, and the proliferation of smooth muscle cells in the vascular wall. Recent experimental evidences suggest that immediate-early (IE) gene and extracellular matrix protein (EMP) gene expression might be closely associated with the attachment, migration, and proliferation of the component cells in the vascular wall and might function in the progression of atherosclerosis. In this study, we examined the mRNA levels of c-fos (an IE gene) and fibronectin (an EMP gene) in aorta and pulmonary trunk of rabbits during the formation of atherosclerotic plaques. New Zealand white rabbits were fed with high cholesterol diet (2%) containing 6% peanut oil for 1, 3 or 6 weeks. Control groups were fed for the same period with normal diet (control Ⅰ) or normal diet containing 6% peanut oil ( control II). At designated time points, different sections of aorta ( arch, thoracic . aorta and abdominal aorta) and pulmonary trunk were excised for RNA isolation and Northern blot analysis. In situ hybridization using digoxigenin-labeled fibronectin and c-fos cDNA probes were conducted to determine the distribution of these mRNAs all arterial segments and those of control II in aortic arch and pulmonary trunk. In contrast, c-fos mRNA levels of aorta were found to be lower in hyperlipidemic group than those in control II. In situ hybridization showed that both fibronectin and c-fos mRNA were highly expressed in the foam cells of the thickened intima of aortic arch and pulmonary trunk of the hyperlipidemic rabbits. These results indicate that fibronectin and c-fos genes are activated during high cholesterol diet induced atherogenesis and suggest that their activation might be involved in the formation of atherosclerotic plaques.



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