本論文主要研究龜鹿二仙膠對於緩解關節與骨骼退化的替代醫療效果，本研 究選用與順天堂科學中藥廠提供的龜鹿二仙膠，利用 DPPH 自由基清除能力測試 檢視龜鹿二仙膠的抗氧化力；再利用化療藥物阿黴素（Doxorubicin, DOX）處理關 節滑膜細胞（HIG-82），藉此誘發關節滑膜細胞的退化與發炎現象，透過細胞存活 率分析法 MTT 分析以及 ELISA 分析，檢視龜鹿二仙膠是否可以緩解化療誘發關 節滑膜細胞（HIG-82）的毒性與發炎；本研究利用化療藥物阿黴素預先處理，藉 此誘發老化小鼠的骨骼疏鬆，然後使用骨骼電腦斷層掃描技術檢視脛骨的骨小樑 與骨密度，藉此評估龜鹿二仙膠是否可以緩解化療藥物阿黴素所誘發小鼠的骨質 疏鬆。本論文結果如圖 7 所顯示，龜鹿二仙膠不但可以顯著增強 DPPH 自由基清 除能力，對於化療藥物阿黴素誘發關節滑膜細胞（HIG-82）的毒性與發炎，也有 顯著的緩解效果；此外，對於化療藥物阿黴素所誘發小鼠的骨質疏鬆，連續餵食 龜鹿二仙膠四週後也有顯著的緩解效果。歸納本研究結果，可以說明龜鹿二仙膠 對於緩解關節與骨骼退化確實具有替代醫療效果。
研究者認為適時給予中高齡長者補充龜鹿二仙膠，將有助於緩解高齡長者行 動不便以及長期臥病在床等問題，大大降低醫療資源與健保經費的龐大支出。
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附錄圖表
圖 1： 全台民眾骨質檢測數據統計分析
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圖 1： 全台民眾骨質檢測數據統計分析
國內知名科學中藥廠–順天堂為民眾進行數十場骨質密度檢測，透過國立臺灣師範大學吳忠信教授與順天堂中藥廠的合作以及數據統計 分析，公佈了臺灣首份「全台民眾骨質危機解密地圖」，檢測結果發現，受測者骨質密度檢測合格的比例僅為 19%，而檢測不合格比例 竟然高達 81%。
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圖 2：龜鹿二仙膠的高效液相層析 3D-HPLC 圖譜
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圖 2：龜鹿二仙膠的高效液相層析 3D-HPLC 圖譜
龜鹿二仙膠主要包含四種中藥材：枸杞（Lycii Fructus）、龜板（Testudinis Carapax et Plastrum）、人參（Ginseng Radix）以及鹿角（Cornu Cervi）。利用高效液相層析 3D-HPLC（200-400 nm，0-65 min）分析龜鹿二仙膠的中藥指紋圖譜，箭頭指示為龜鹿二仙膠的活性指標物 質。
32
圖 3：龜鹿二仙膠的 DPPH 清除氧化自由基能力測定
DP PH
33
圖 3：龜鹿二仙膠的 DPPH 清除氧化自由基能力測定
（圖 A） 利用 DPPH 方法對順天堂龜鹿二仙膠進行抗氧化能力測定，結果顯示龜鹿二仙膠具有清除自由基的抗氧化功效。（圖 B）統 計結果顯示龜鹿二仙膠隨著測試濃度增加，清除自由基的抗氧化效率顯著增加。各組數值以平均值±平均值標準誤差（Standard error of the mean, SEM）表示，且用單因子變異數分析（One-way analysis of variance, ANOVA）以及 S-N-K 多重差距檢定 （Student-Newman-Keuls multiple range test ）來分析是否有顯著誤差，若有顯著誤差則用**p<0.01 以及*p<0.05 表示。
34
圖 4：龜鹿二仙膠緩解脂多醣（LPS）誘發關節滑膜細胞（HIG-82）活性
脂多醣處理組
35
圖 4：龜鹿二仙膠緩解脂多醣（LPS）誘發關節滑膜細胞（HIG-82）活性
（圖 A） 利用 MTT 方法對順天堂龜鹿二仙膠之抗細胞活性測定，統計結果顯示：HIG-82 關節滑膜的細胞存活率會隨著龜鹿二仙膠處 理濃度增加而顯著增加。（圖 B） 而添加脂多醣（LPS）誘發關節滑膜細胞（HIG-82）損傷，雖然細胞存活率明顯減少，也會隨著龜鹿 二仙膠處理濃度增加而顯著增加。這結果說明龜鹿二仙膠確實具有保護肝臟巨噬細胞損傷的功效。各組數值以平均值±平均值標準誤差 表示，且用單因子變異數分析以及 S-N-K 多重差距檢定來分析是否有顯著誤差，若有顯著誤差則用**p<0.01 以及*p<0.05 表示。
36
圖 5：龜鹿二仙膠緩解化療誘發關節滑膜細胞（HIG-82）發炎
37
圖 5：龜鹿二仙膠緩解脂多醣（LPS）誘發關節滑膜細胞（HIG-82）發炎因子
（圖 A） 利用酵素結合免疫吸附（ELISA）分析，可觀察到添加脂多醣（LPS）誘發關節滑膜細胞（HIG-82）發炎的 TNF-α發炎指數 較空白處理組顯著增加（p<0.01），當添加龜鹿二仙膠，於 72 小時檢測關節滑膜細胞（HIG-82），可以發現 TNF-α 發炎指數顯著降低
（圖 A） 利用酵素結合免疫吸附（ELISA）分析，可觀察到添加脂多醣（LPS）誘發關節滑膜細胞（HIG-82）發炎的 TNF-α發炎指數 較空白處理組顯著增加（p<0.01），當添加龜鹿二仙膠，於 72 小時檢測關節滑膜細胞（HIG-82），可以發現 TNF-α 發炎指數顯著降低