臺灣博碩士論文加值系統

大白鼠的旁血管脂肪組織 perivascular adipose tissue (PVAT) 會釋放出一種使血管平滑肌放鬆的物質，這個物質到現在還未被確認。本實驗的目的要確認旁血管脂肪組織衍生之舒張因子 perivascular adipose tissue-derived relaxing factor (PVATRF) 。本研
究將 Wistar Kyoto (WKY) 公鼠分成對照組 (含PVAT的主動脈環, aortic rings+PVAT) 以及實驗組 (無PVAT的主動脈環, aortic ring-PVAT) 。於組織皿中， phenylephrine (Phe, 1 nM － 10 μM)及 norepinephrine (NE, 1 nM － 10 μM) 使 aortic rings-PVAT 的收縮比使aortic rings+PVAT 的收縮為大。 KCl (10 － 100 mM) 引起的收縮，則
aortic rings-PVAT 或 +PVAT 都是沒有差別的。在 superfusion bioassay cascade 中，流經 PVAT 的 Krebs’ solutions 會導致主動脈環 (去除內皮及PVAT) 放鬆。收集會放鬆主動脈環之液體，用 GC-MS 分析的結果顯示液體中含有 palmitic acid methyl ester (PAME) 。流經去除內皮及 PVAT 的主動脈環不會造成主動脈環放鬆，也不會釋放PAME 。 PVATRF 跟 PAME 的釋放都是 Ca2+-dependent 。外給 PAME 依 concentration-dependent 的方式引起主動脈環放鬆，其 EC 50 為 3.32 pM 。上述PVATRF 跟PVAT 引起血管放鬆不是透過內皮， 而且會被4-aminopyridine (4-AP, 2 mM) 與tetraethylammonium (TEA, 5 and 10 mM)所抑制，然而卻不會被 TEA (1, 3 mM), glimenclamide (3 μM), iberiotoxin (100 nM), miconazole (60 μM), SFK-525A (10 μM), or indomethacin (10 μM) 。據此推測 PAME 是一種 PVATRF 。

The perivascular adipose tissue (PVAT) from the rat aorta has been shown to release yet to be determined substance(s) that relaxes the medial smooth muscle cells. The aim of this study was to determine the nature of the PVAT-derived relaxing factor (PVATRF). The control (+PVAT) and experimental (-PVAT) aortic rings of male Wistar Kyoto (WKY) rats were examined using both in vitro tissue bath and superfusion bioassay cascade techniques. In the in vitro tissue bath study, contractile responses to phenylephrine (Phe, 1 nM - 10 μM) and norepinephrine (NE, 1 nM - 10 μM) were significantly greater in aortic rings-PVAT than aortic rings+PVAT. KCl (10 - 100 mM)-induced aortic constrictions, however, were not different between aortic rings+PVAT and aortic rings-PVAT. In superfusion bioassay cascade study, the Krebs’ solution that had superfused through PVAT preparations resulted in relaxation of aortic rings-PVAT (denuded of endothelial cells/EC) down the cascade. The perfusates resulting in aortic dilation were collected and analyzed by GC-MS. The results indicated the presence of significant concentrations of palmitic acid methyl ester (PAME) in the perfusates. Superfusing smooth muscle (namely the aortic ring without EC and PVAT) did not cause any aortic relaxation or PAME release. Releases of PVATRF and PAME were Ca2+-dependent. Exogenous PAME in a concentration-dependent manner induced aortic relaxation with EC50 value of 3.32 pM. The aortic relaxations induced by PVATRF and PAME were independent of the endothelium, and were blocked by 4-aminopyridine (4-AP, 2 mM) and tetraethyl ammonium (TEA, 5 and 10 mM). The relaxations, however, were not affected by TEA at 1, 3 mM, glimenclamide (3 μM), iberiotoxin (100 nM), miconazole (60 μM), SKF-525A (10 μM), or indomethacin (10 μM). These results suggest that PAME is a PVATRF.

一、 英文摘要…………………………………………………………05
二、 中文摘要…………………………………………………………06
三、 背景介紹…………………………………………………………07
四、 理論根據與研究目的……………………………………………08
五、 實驗材料與方法…………………………………………………10
六、 實驗流程…………………………………………………………16
七、 結果………………………………………………………………19
八、 討論………………………………………………………………22
九、 參考文獻…………………………………………………………24
十、 結果圖……………………………………………………………27

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