肥胖加速肌肉流失的發生，故本實驗探討在高脂飲食下給予魚油對肌肉流失之影響。魚油在人體實驗已證實可以增加肌肉蛋白質合成，但動物實驗研究相對缺乏，因此機制不明。先前本研究室於高脂飲食中添加 5% 魚油，8 週後發現其肌肉組織的 AMPKα、PGC-1α、PPARγ 及 LPL 表現量顯著增加，抑制高脂飲食造成的骨骼肌脂肪酸堆積，另外，魚油也透過顯著降低FOXO3a 來阻擋下游肌肉萎縮基因 Atrogin-1 及 MuRF1 表現，改善肌肉流失。然而，魚油對於改善高脂飲食所造成的肌肉流失機制仍不夠完整。本研究以 5 週齡雄性 SD 為實驗動物，共分 3組: (1) 正常飲食 (Normal control diet, NC) (2)高脂飲食 (High fat diet, HF; 15% lard) (3) 魚油組 (HF 10% lard + 5% fish oil, FO)。實驗共進行 16 週。結果顯示，魚油可以降低血漿 TC、TG 及肝臟 TG。大鼠、腓腸肌、伸趾長肌脛骨前肌及比目魚肌組織切片顯示，高脂飲食組的肌肉纖維截面積相較於正常控制組和魚油組顯著下降 (p < 0.05)，且在肌肉纖維截面積分布上，高脂飲食組的肌肉纖維截面積相較於正常控制組，其肌肉纖維截面積往小移動，而魚油組的肌肉纖維截面積相較於高脂飲食組，其肌肉纖維截面積往大移動。但肌肉重量卻沒有顯著差異。魚油透過活化 p-AKT/AKT、p-mTOR/ mTOR 途徑，增加蛋白質合成，並抑制 FOXO 和 NFκB 進入細胞核中轉錄 Atrogin-1 及 MuRF1，減少肌肉降解。同時魚油會增加 p-AMPKα/AMPKα、PGC-1α 增加粒線體生合成及降低 PINK1，改善高脂飲食所造成的粒線體失能。此外，魚油透過降低 PPARγ，減少肌肉中脂肪酸堆積；增加 p-AMPKα/AMPKα、PGC-1α，增加肌肉中脂肪酸氧化。由於肌肉中的脂肪含量降低，抑制了發炎因子 TNF-α、NFκB，並降低 p62 自噬因子。綜合上述結果，推測長期攝食魚油有助於減緩高脂飲食所造成的肌肉品質不良。

Obesity maybe result in muscle wasting. The study was designed to investigate the effect of fish oil fed with high fat diet on muscle wasting. Fish oil has been confirmed for increased protein synthesis in human study, but it is lack of animal study. In present study, we suggested fish oil suppress lipid accumulation in skeletal muscle via AMPKα/PGC-1α/LPL/PPARγ pathway. Moreover, fish oil significantly decreased FOXO3a and inhibit MuRF1/Atrogin-1 expression to improve muscle wasting fed with high fat diet. Nevertheless, the mechanism of fish oil improved muscle wasting is not clear. In this study, rats are divided into three groups: (1) Normal control diet (NC). (2) High fat diet (HF). (3) High fat diet with 5% fish oil (FO). The experiment was done for 16 weeks. Results showed that 5% fish oil significantly decreased plasma TC, TG and liver TG. Histology of cross sectional area upon gastrocnemius muscle, extensor digitorum longus muscles, tibialis anterior muscles and soleus muscles. These demonstrate decrease cross-sectional areas of muscle fiber in high fat diet less than normal control diet and fish oil diet. The frequency of cross sectional area with high fat diet reduce to normal control diet, and fish oil diet can enhance the frequency of cross sectional area. However, not significant differences in skeletal muscle mass. Fish oil promoted protein synthesis via p-AKT/AKT, p-mTOR/ mTOR. Fish oil also prevented nuclear translocation of FOXO1a and NFκB, thereby suppressing MuRF1and Atrogin-1. In addition, fish oil upregulated p-AMPKα/AMPKα, PGC-1α to enhance mitochondrial biogenesis and downregulated PINK1 to improve mitochondrial dysfunction. Futhermore, fish oil decreased fatty acid accumulation via PPARγ and increase fatty acid oxidation via p-AMPKα/AMPKα, PGC-1α. Through reduced lipid accumulation in skeletal muscle, reduced inflammation mediators TNF-α and NFκB and inhibited autophagy by decreasing the level of p62. In conclusions, our results demonstrated fish oil as a supplement for alleviating bad muscle quality in high fat fed rats.

摘要----------------------------------------------------II
Abstract ----------------------------------------------III
目錄---------------------------------------------------III
圖目錄---------------------------------------------------V
表目錄-------------------------------------------------VII
第一章 前言--------------------------------------------1
第二章 文獻回顧----------------------------------------3
第一節、肌少症 -------------------------------------------3
第二節、高脂飲食------------------------------------------5
第三節、膳食魚油 -----------------------------------------6
第四節、肌肉合成與降解-------------------------------------8
第五節、粒線體-------------------------------------------10
第六節、發炎介質 ----------------------------------------11
第七節、脂質代謝基因蛋白----------------------------------12
第八節、粒線體基因蛋白------------------------------------16
第九節、肌肉代謝基因蛋白----------------------------------17
第十節、肝臟代謝基因蛋白----------------------------------21
第三章 實驗動機及目的---------------------------------22
第一節、實驗動機-----------------------------------------22
第二節、實驗流程-----------------------------------------22
第三節、分析項目 ----------------------------------------24
第四章 實驗材料---------------------------------------25
第一節、實驗動物 ----------------------------------------25
第二節、實驗飼料 ----------------------------------------25
第三節、膳食魚油-----------------------------------------25
第四節、實驗儀器 ----------------------------------------25
第五章 實驗方法---------------------------------------27
第一節、膳食魚油脂肪酸組成分析----------------------------27
第二節、脂肪酸分析---------------------------------------27
第三節、動物分組-----------------------------------------27
第四節、動物飼養及樣品收集--------------------------------28
第五節、飼料配方-----------------------------------------28
第六節、樣品分析-----------------------------------------29
1.血漿分析-----------------------------------------------29
2.肝臟分析-----------------------------------------------31
3.糞便分析-----------------------------------------------32
4.蛋白質分析-西方墨點法----------------------------------33
5.石蠟切片及染色-----------------------------------------35
6.統計分析-----------------------------------------------36
第六章 結果-------------------------------------------37
第一節、魚油對大白鼠體重、攝食量及血液中瘦素之影響----------37
第二節、魚油對大白鼠臟器之影響----------------------------37
第三節、魚油對大白鼠肌肉重量及肌肉面積之影響---------------37
第四節、魚油對大白鼠血漿、肝臟、糞便生化指標之變化----------38
第五節、魚油對大白鼠肌肉脂質代謝分子機制之影響-------------38
第六節、魚油對大白鼠蛋白質合成分子機制之影響---------------39
第七節、魚油對大白鼠蛋白質降解分子機制之影響---------------39
第八節、魚油對大白鼠粒線體生合成之影響---------------------40
第九節、魚油對大白鼠肝臟脂質代謝之影響---------------------40
第七章 討論-------------------------------------------41
第一節、魚油對體重、攝食量、組織臟器的影響-----------------41
第二節、魚油對大白鼠肝功能及發炎因子的影響-----------------42
第三節、魚油對大白鼠蛋白質合成分子機制之影響---------------44
第四節、魚油對大白鼠蛋白質降解分子機制之影響---------------44
第五節、魚油建議攝取量------------------------------------46
第八章 結論-------------------------------------------47
第九章 參考文獻---------------------------------------49

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