臺灣博碩士論文加值系統

中文摘要 I
ABSTRACT II
謝誌 IV
目錄 V
圖表目錄 IX
第一章 前言 1
第二章 文獻回顧 2
2.1 骨碎補 2
2.2 骨碎補成分及生理活性 4
2.3 骨質吸收及重建影響因子 5
2.4 骨母細胞 （osteoblasts） 生理活性 10
第三章 材料與方法 13
3.1 材料 13
3.2 儀器設備 13
3.3 化學試藥 14
3.4 實驗設計 16
3.5 驗方法 20
3.5.1大葉骨碎補組織切片鑑定 20
3.5.2大葉骨碎補水萃取物製備 24
3.5.3大葉骨碎補乙醇萃物製備 24
3.5.4前骨母細胞MC3T3-E1培養及保存 24
3.5.5前骨母細胞MC3T3-E1細胞生長測定 25
3.5.6鹼性磷酸酶活性測定 25
3.5.7西方墨點法測試BMP-2, ALP,Col-1, Runx-2蛋白表現量 28
3.5.8骨細胞礦化節染色及測定 30
3.5.9 Diaion HP20 管柱區分 30
3.5.10 Sephadex LH20 管柱區分 30
3.5.11析出 31
3.5.12 MCI-gel CHP20P管柱區分 31
3.5.13薄層層析分析thin layer chromatography (TLC) 31
3.5.14 DFW及CP製備 31
3.5.15 C57BL/6JNarl 小鼠飼養 32
3.5.16劑量分組 34
3.5.17檢體採集 34
3.5.18 Osteocalcin（OCN）之含量測定法 35
3.5.19 TNF−α之含量測定法 35
3.5.20 IL-1β之含量測定法 36
3.5.21蘇木紫/伊紅染色法（ Hematoxylin and Eosin stain ； H/E stain ） 36
3.5.22統計分析 36
第四章 結果與討論 37
4.1細胞試驗結果 37
4.1.1大葉骨碎補組織切片鑑定結果 37
4.1.2第一部大葉骨碎補水萃取物製備結果 39
4.1.3大葉骨碎補乙醇萃取物製備結果 39
4.1.4大葉骨碎補水及乙醇萃取物對前骨母細胞MC3T3-E1細胞生長影響 39
4.1.5大葉骨碎補水及乙醇萃取物對前骨母細胞MC3T3-E1鹼性磷酸酶活性影響 42
4.1.6大葉骨碎補水及乙醇萃取物對前骨母細胞MC3T3-E1之BMP-2, ALP,COL-1, Runx-2蛋白質表現影響 44
4.1.7大葉骨碎補水及乙醇萃取物對前骨母細胞Runx-2蛋白質表現影響 44
4.1.8大葉骨碎補水及乙醇萃取物對前骨母細胞BMP-2蛋白質表現影響 47
4.1.9大葉骨碎補水及乙醇萃取物對前骨母細胞Col-1蛋白質表現影響 49
4.1.10 大葉骨碎補水及乙醇萃取物對前骨母細胞ALP蛋白質表現影響 51
4.1.11大葉骨碎補水及乙醇萃取物對前骨母細胞MC3T3-E1骨質礦化影響 53
4.2純化實驗結果 55
4.3動物實驗結果 69
4.3.1老鼠體重變化 69
4.3.2 介白素1β (Interleukin 1 beta (IL-1β)) 及腫瘤壞死因子α (Tumor necrosis factor alpha (TNF-α)) 71
4.3.3鹼性磷酸酶 (Alkaline phosphatase (ALP)) 及骨鈣素 (Osteocalcin (OCN)) 測定 75
4.3.4左股骨骨小樑之組織染色 (H/E stain)測定 78
第五章 結論 80
第六章 參考文獻 83
第七章 作者簡介 92

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