在已開發國家中發現越來越多影響生殖系統的問題出現，除了生活壓力外，晚婚社會、飲食與生活習慣的改變，均影響了男性生殖器官發育、精子活動力及數目。現今人們追求精緻可口的食物，但已有文獻證實高脂飲食會造成肥胖、高血脂、高血壓及糖尿病等與代謝症候群相關的疾病。高脂飲食會促使細胞長期浸潤在油脂中，而產生發炎反應及增加細胞膜上的磷脂質氧化壓力造成氧化傷害，而因為睪丸外膜富含多元不飽和脂肪酸，更加容易引起氧化傷害，影響睪丸功能和精子品質。近年來非常熱門的熱量限制議題中，許多學者相繼投入對於精子影響的研究，但是熱量限制對於睪丸功能的影響依然存有爭議性。故本研究擬探討高脂飲食和熱量限制飲食對於老化過程中睪固酮分泌及睪丸組織氧化壓力產生的影響。研究使用四、九和十六月齡的Sprague-Dawley (SD) 大鼠，隨機分成三組，給予標準飼料、高脂飼料或標準飼料並加以熱量限制，餵養十四週後犧牲，採集血液及睪丸，以酵素免疫分析方法 (enzyme immunoassay，EIA)測量血漿中睪固酮濃度，並以西方墨點法 (Western blot)分析steroidogenic acute regulatory protein (StAR protein)、Aquaporin 7的蛋白表現，以化學套組分析抗氧化酵素的活性及脂質過氧化物(TBARS)產量，另外取一部分睪丸用以包埋切片，計算睪丸組織內曲細精管的管腔直徑及觀察型態的變化。所有的實驗結果以平均值標準誤 (meanS.E.M.) 表示，以SPSS 18.0 進行統計分析。先以one way analysis of variance (one-way ANOVA) 進行資料分析處理，並以Duncan’s post-hoc 測驗法事後檢定組間的平均值的差異，以p<0.05視為具有顯著差異。曲細精管型態則以交叉表進行分析。結果顯示飼料的介入並不會影響到睪丸組織、副睪組織重量，但經過體重加權後高脂飼料組的比值和熱量限制組相比有下降的趨勢。在抗氧化指標的部分雖然沒有統計上差異，但是熱量限制組的抗氧化指標和高脂飼料組相比仍有較高的趨勢。血清中睪固酮濃度雖然組別上沒有統計差異，但是熱量限制過後，大鼠血清中睪固酮平均濃度和高脂飼料相比有較高的趨勢。除此之外，在曲細精管型態上，老年大鼠經過熱量限制後和高脂飼料組相比，較能維持曲細精管的正常型態。蛋白質表現的部分，老年大鼠經過熱量限制後其睪丸組織中StAR、Aquaporin 7蛋白表現量增加。整體來說，熱量改變與年齡對生殖系統均有影響，熱量限制雖然會降低生殖相關組織的重量，反之增加SOD活性及StAR表現，而年齡增加造成曲細精管型態改變可能與glutathion合成減少有關。

More and more problems, including stress late marriage and changes in the diet and lifestyle affected the reproductive system, motility and number of sperm. High-fat diet has been confirmed to cause obesity, high cholesterol, hypertension, diabetes and metabolic syndrome. Cells long-term soaked in lipid induce inflammation and increase oxidative stress. The cellular membrane of testicular tissue is rich in polyunsaturated fatty acids, and likely to cause oxidative stress and then damage testicular function and sperm quality. There are many researchers focused on the effect of caloric restriction on testis, but the results still are controversial. Therefore, this study was to investigate the impacts of high-fat diet and calorie-restricted diet on testis, including oxidative stress, testosterone release and arrangement of seminiferous tubules during the aging process. In this study, four, nine and sixteen-month-old Sprague-Dawley (SD) rats were randomly divided into three groups and given a standard diet, high fat diet or a standard diet with caloric restriction for fourteen weeks. After sacrifice, blood was collected, and subjected to enzyme immunoassay methods (enzyme immunoassay, EIA) to measure plasma testosterone levels. Western blot was used to measure the expression of steroidogenic acute regulatory protein (StAR protein) and aquaporin 7 protein. The activities of antioxidant enzymes and productions of lipid peroxidation (TBARS) and glutathione were measured with commercial kits. The testicular tissues were embedded, and stained with hematoxylin & eosin to analyze the diameter and arrangement of seminiferous tubules. All experimental results were presented as mean standard error mean (mean SEM) and used the software SPSS 18.0 for statistical analysis. The treatment means were tested for homogeneity by one way analysis of variance (one-way ANOVA) and the difference between specific means was tested for significance by Duncan's post-hoc test. A difference between two mens was considered statistically significant when p<0.05. The results showed that the intervention of diet does not affect the weight of testicular tissues and epididymal tissues. There were no difference on oxidative stress and plasma testosterone release, but calorie restriction increased the plasma testosterone release compared to high fat diet. In the seminiferous tubules, the rating score represented a decreased pattern in caloric restriction compared the seminiferous tubules situation worse, aged rats after caloric restriction and high fat diet group compared with high-fat diet during aging. The protein expression of StAR and aquaporin 7 were also increased in aged combined with caloric restriction, but not in aged combined with high-fat diet. Overall, both of energy the change with and aging affect the reproductive system. Calorie restriction, reduced the weight of reproductive organs, increase the reactivity of SOD and StAR protein expression. The disarrangement of seminiferous tubules during ageing may be due to decreased glutathione production.

第壹章 緒論…………………………………………………………1
第一節 研究背景……………………………………………….1
第二節 研究假設…………………………………………….....2
第三節 研究目的……………………………………………….2
第貳章 文獻探討……………………………………………………3
第一節 睪丸的介紹 …………………………………………….3
第二節 氧化壓力………………………………………………..6
第三節 熱量限制………………………………………………..9
第四節 高脂飲食………………………………………………..10
第五節 老化……………………………………………………..12
第參章 研究方法…………………………………………………….15
第一節 實驗動物飼養與處理…………………………………..15
第二節 實驗分組與流程………………………………………..15
第三節 實驗材料與樣本分析…………………………………..16
第四節 資料統計及分析………………………………………..28
第肆章 實驗結果…………………………………………………….29
第一節 不同飼料介入對於不同年齡大鼠體重及組織重量的影響……………………………………………………..29
第二節 不同飼料介入對於不同年齡大鼠睪固酮分泌及曲細精管的影響………………………………………………..30
第三節 不同飼料介入對於不同年齡大鼠抗氧化指標的影響……………………………………………………..31
第四節 不同飼料介入對於不同年齡大鼠睪丸蛋白表現的影響……………………………………………………..31
第伍章 討論………………………………………………………….33
第一節 不同飼料介入對於不同年齡大鼠體重、睪丸組織等的影響………………………………………………………..33
第二節 不同飼料介入對於不同年齡大鼠睪固酮分泌的影響..33
第三節 不同飼料介入對於不同年齡大鼠曲細精管的影響…..34
第四節 不同飼料介入對於不同年齡大鼠睪丸組織抗氧化指標的影響……………………………………………………..35
第五節 不同飼料介入對於不同年齡大鼠睪丸蛋白表現的影響………………………………………………………..35
第陸章 結論………………………………………………………….38
參考文獻……………………………………………………………...39
圖表說明……………………………………………………………...54

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