目的：本研究旨在探討低蛋胺酸飲食與耐力運動對成長中雄性大鼠骨骼之代謝、組織型態與生物力學特性之影響。
方法：週齡7週之雄性SD大鼠分為六組（每組n=12），依照飲食中蛋胺酸（methionine, MET）含量與運動訓練 (exercise, EXE) 之有無，分成0.86%MET、0.52%MET、0.17%MET、0.86%MET+EXE、0.52%MET+EXE、0.17%MET+EXE等六組，其中，以0.86%MET為控制組，而0.52%MET組和0.17%MET組分別為降低40%與80%MET含量之低蛋胺酸飲食組別；耐力運動組別的動物則以24公尺/分鐘、60分鐘/天及5天/週之跑步訓練介入。實驗介入期為期八週。採取自由取食之方式，每週記錄兩天動物之體重變化與飲食飲水量，完成為期八週之介入後，將犧牲動物後所得之血液與骨骼樣本，進行血清骨代謝、骨組織型態和骨組織生物力學特性等變項分析，統計方法為二因子（蛋胺酸耐力運動）變異數分析(two-way ANOVA) (α = .05)。
結果：經過八週不同蛋胺酸飲食與耐力訓練後，體重方面，METEXE之交互作用達顯著水準 (p=.003)，0.52%MET組之大鼠在體重增加方面顯著高於0.86%MET組和0.17%MET組，又0.86%MET組顯著高於0.17%MET組，而運動組顯著低於非運動組。在骨代謝的血液分析方面，本研究分別測量骨鈣素 (osteocalcin)及第一型膠原羧基端肽（C-telopeptide fragments of collagen type 1, CTX-1）分別代表骨生成指標及骨吸收指標，此外，本研究亦分析血中類胰島素生長因子-1 (insulin-like growth factor-1, IGF-1)及類胰島素生長因子連結蛋白-3（insulin-like growth factor binding protein-3, IGFBP-3）以作為發展與老化指標之用，結果顯示，各項指標均在MET或EXE之主要效果呈現顯著水準，亦有部份指標在交互作用達顯著，低蛋胺酸飲食及運動均對上述指標有不同呈度的向下調節作用。在骨靜態組織型態方面，攝食0.17%MET之大鼠在各項海綿骨結構參數較低於0.86%MET組和0.52%MET組，而運動之大鼠則呈現較高於非運動組之骨小樑厚度 (p<.001)及較低於非運動組之破骨細胞數。在動態組織型態分析顯示，骨礦化與生成率方面均為0.17%MET飲食之動物相較於0.86%MET和0.52%MET者顯著較低，另運動組較非運動組低。結構性生物力學部分，0.17%MET飲食者有顯著較低之抗彎曲負荷、抗彎曲能量吸收及硬度，然在材料層次的生物力學特性方面，0.17%MET飲食者之降伏點應力值 (p=.002) 和楊氏係數 (p=.001) 則顯著較高。
結論：降低40%蛋胺酸含量的低蛋胺酸飲食對於骨骼各方面之影響無異於正常飲食，而降低80%蛋胺酸含量相較之下似乎呈現較差的骨骼型態與結構性生物力學特性，然而，在老化相關的血液指標與骨組織之材料層次生物力學特性方面，低蛋胺酸飲食則呈現其優勢，此外，若分別佐與運動，則對部份參數亦有助益。

Abstract
Purpose: To investigate the effects of low methionine (MET) diets and endurance exercise (EXE) on bone metabolism, histomorphometry and biomaterial properties in growing male rats.
Methods: Seventy-two male SD rats (7 weeks old) were randomly assigned to six groups, which were the 0.86%MET, 0.52%MET, 0.17%MET, 0.86%MET+EXE, 0.52%MET+EXE and 0.17%MET+EXE groups (n=12 for each). Control animals were fed with a purified diet containing 0.86% methionine. In addition, the 0.52% and 0.17% MET-fed animals were set as the low MET diet groups. For the EXE groups, animals were subjected to treadmill running training by a protocol of 24 m/min, 60 min/day and 5 days/week for 8 weeks. Body weights and food intake of animals were measured twice a week. After the end of eight-week intervention, all animals were sacrificed under deep anesthesia. Samples of blood and bone were collected and stored for serum bone metabolism, histomorphometry and tissue level biomechanical properties analysis. Statistical analysis was processed by using two-way （METEXE）ANOVA (α = .05).
Results:
In body weight, main effects of EXE and MET, and interaction all reached significant level (post hoc comparison: the 0.52%MET group> the 0.86%MET group > the 0.17%MET group; and each exercise group was significantly lower than its parallel non-exercise group). In serum marker assays, we measured osteocalcin, CTX-1, IGF-1 and IGFBP-3 as indexes of bone metabolism and development/aging related markers. Significance of MET main effects, EXE main effects or interaction were shown, respectively, in those serum markers. Low-MET diet and exercise show different down-regulated effects on those markers. In histomorphometry, the main effect analysis of MET showed that BVTV and trabecular number were significantly lower in the 0.17%MET-fed rats. And, trabecular separation of the 0.17%MET rats was significantly higher than that of the 0.86%MET and 0.52%MET rats. Additionally, in the main effect of EXE, the exercise-trained rats were higher and lower than the non-EXE rats in trabecular thickness and osteoclast number, respectively. In dynamic histology analysis, the 0.17%MET rats showed significantly lower bone formation (MS/BS and BFR/BS) compared to the 0.86%MET and 0.52%MET rats. And, the exercise-trained rats showed lower MS/BS and BFR/BS as compared to the non-exercise rats. In biomaterial properties, femora of the 0.17%MET rats were significant lower in fracture load, yield load, stiffness and post-yield energy as compared to the 0.86%MET and 0.52%MET rats. Conversely, the 0.17%MET rats revealed significant increase in yield stress and Young’s modulus.
Conclusion:
There was no negative effect of the 40% methionine reduction on bone metabolism, histomorphometry and biomaterial properties. It seemed that the 80% methionine diet caused an inferior architecture in spongy bone and a decreased structural strength in cortical bone. However, in aging-related serum markers and material level biomechanical properties, low-MET diet shown its benefit. And, endurance exercise intervention showed modifications for parameters investigated in the current study.

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