臺灣博碩士論文加值系統

　　骨關節炎是全世界最常見的慢性疾病之一，其最主要的症狀是疼痛，傳統止痛藥包括類固醇與非類固醇抗發炎藥，長期使用會有消化性潰瘍、下肢水腫及腎功能損傷等副作用，藥物治療的副作用非常令人憂慮，因此，研究自然界中可對抗關節發炎的天然物質更顯得重要許多。本研究實驗將三種成份—（a）有機大麥若葉苗(Barley grass)萃取物。（b）山竹(Garcinia mangostana)萃取物。（c）天然蔬果酵素(Natural vegetable fruit enzymes )，分別以各樣品之水萃取液，處理3T3老鼠纖維母細胞或細胞培養液，並進行基質金屬蛋白酶與透明質酸酶的酵素活性抑制分析及DPPH自由基清除能力分析。
　　實驗結果顯示：1.0%山竹水萃取液處理24與48小時後，對MMP-2、MMP-9及MMP-13抑制活性之能力開始顯見，另以2.5%山竹水萃取液處理24與48小時後， 發現從24小時開始對MMP-2、MMP-9及MMP-13有顯著抑制活性之能力，當濃度提高至5.0%及10.0%時，山竹水萃取液則可完全抑制MMP-2、MMP-9及MMP-13活性之能力。相對以5.0%、10.0%之大麥若葉與天然蔬果酵素水萃取液處理細胞24與48小時後，對MMP-2、MMP-9及MMP-13並無明顯之抑制能力。而1.0%之山竹水萃取液在處理時間為24小時，即可以完全抑制透明質酸酶之活性，顯示其具有極高的抑制能力；最終濃度5.0%的天然蔬果酵素水萃取液於處理時間24小時即具有些許抑制透明質酸之效果，當最終處理濃度提高至10.0%時且時間延長至48小時，亦具有強的抑制透明質酸酶之效果；而大麥若葉水萃取液於最終濃度為5.0%及10.0%，在24小時時對於透明質酸酶具有些許的抑制效果，在48小時則與控制組相比無明顯差異，即不具有抑制透明質酸酶作用的能力。而另抗氧化實驗結果顯示，山竹濃度200 μg/mL以上 有很高DPPH自由基清除率達88%；而大麥若葉以濃度400 μg/mL其DPPH自由基清除率為17%；當濃度為400 μg/mL時，天然蔬果酵素則不具DPPH自由基清除力。綜合以上研究數據，山竹兼具有抑制MMPs及抑制透明質酸酶之抗發炎能力與抗氧化能力，為一種可應用於關節發炎的有效成份。

　　The primary symptom of Osteoarthritis is pain, which is one of the most common chronic diseases in the world as well. Traditional painkillers (including steroid and non-steroid anti-inflammatory drugs) can cause side effects under long term use, such as peptic ulcer, lower limb edema and impairment of kidney. Since the side effects of drug treatment are quite disturbing annoying, the study of natural substances that which can fight joint inflammation appears especially important. This study used: (a) barley grass extract, (b) Garcinia mangostana extract, and (c) natural vegetable fruit enzymes, or their mixtures, applied with 3T3 mouse fibroblast or cell supernatant, and proceeded with the analysis of enzyme activities of matrix metalloproteinases and hyaluronidaseas as well as DPPH free-radical scavenging capacity.
　　The experimental results show that the ability of inhibition towards MMP-2, MMP-9 and of MMP-13 was obvious by treating 1.0% mangosteen water extracted for 24 and 48 hours, whereas 2.5% of Garcinia mangostana extract showed apparent inhibition on the activity of MMP-2, MMP-9 and MMP-13 after 24 h treatment (in the 24 and 48 h treatment). Comparatively speaking, barley grass extract and natural vegetable fruit enzymes , at concentrations of 5.0% and 10.0%, show unapparent inhibition on MMP-2, MMP-9 and MMP-13 in 24 and 48 h after treatment. 1.0% of the mangosteen water extract of the processing time of 24 h, completely inhibited the activity of hyaluronidase, showing its high rejection; a final concentration of 5.0% for a long time Secretary natural vegetable fruit enzymes water extract of the processing time 24 h has little effect of inhibition of hyaluronic acid; nevertheless, when the concentration was increased to 10.0% and the time to 48 h, a strong effect of inhibition of hyaluronidase has been observed. barley water extract at a final concentration of 5.0 and 10.0% for hyaluronidase has little inhibition effect at 24 h; however, with 48 h compared with the control group which does not have the ability to inhibit the role of hyaluronidase.
　　Antioxidant experimental result indicates that the DPPH free-radical scavenging rate of 200 μg/mL of Garcinia mangostana is quite high, i.e., up to 83.0%; while that of 400 μg/mL of barley grass is 17.0%; and 400 μg/mL of natural vegetable fruit enzymes showed no apparent DPPH free-radical scavenging capability.
　　Garcinia mangostana possesses the ability to inhibit MMPs and hyaluronidaseas, as well as the anti-inflammatory and antioxidative properties based on the above experimental data. In conclusion, Garcinia mangostana is an active ingredient that can be used for the treatment of joint inflammation.

目錄
中文摘要 ---------------------------------------------------------------------------------- I
表目錄 ----------------------------------------------------------------------------------- IX
圖目錄 ------------------------------------------------------------------------------------ X
縮寫表 ---------------------------------------------------------------------------------- XII
第一章 緒論 ------------------------------------------------------------------------------ 1
1-1. 前言 ----------------------------------------------------------------------------- 1
1-2. 研究動機與目的 -------------------------------------------------------------- 2
1-3. 研究架構 ----------------------------------------------------------------------- 3
第二章 文獻回顧 ------------------------------------------------------------------------ 4
2-1. 植物介紹 ----------------------------------------------------------------------- 4
2-1-1. 山竹 --------------------------------------------------------------------- 4
2-1-2. 巴西酵素 --------------------------------------------------------------- 7
2-1-3. 大麥若葉 --------------------------------------------------------------- 8
2-2. 關節炎 -------------------------------------------------------------------------- 9
2-2-1. 關節構造 -------------------------------------------------------------- 11
2-2-2. 基質金屬蛋白酶 ---------------------------------------------------- 12
2-2-3. 透明質酸酶 ---------------------------------------------------------- 14
2-3. 抗氧化 ------------------------------------------------------------------------ 19
第三章 研究材料、儀器與方法 ---------------------------------------------------- 23
3-1. 研究材料 --------------------------------------------------------------------- 23
3-1-1. 植物來源 ------------------------------------------------------------- 23
3-1-2. 藥品試劑 ------------------------------------------------------------- 24
3-2. 實驗方法 --------------------------------------------------------------------- 27
3-2-1. 植物之水萃取法 ---------------------------------------------------- 27
3-2-2. 細胞活化與繼代培養 ---------------------------------------------- 27
3-2-2-1. 細胞來源及培養基組成 ------------------------------------ 27
3-2-2-2. 細胞解凍活化 ------------------------------------------------ 28
3-2-3. 體外動物細胞存活率分析 ---------------------------------------- 28
3-2-4. 基質金屬蛋白酶活性抑制分析 ---------------------------------- 29
3-2-5. 透明質酸酶活性抑制分析 ---------------------------------------- 30
3-2-6. DPPH 自由基清除能力分析 --------------------------------------- 31
第四章 結果與討論 ------------------------------------------------------------------- 33
4-1. 以山竹、久司巴西酵素及大麥若葉水萃取液處理 3T3 纖維母細胞
之細胞存活率分析 --------------------------------------------------------------- 33
4-3. 山竹、久司巴西酵素及大麥若葉水萃取液之 MMP-2、MMP-9 及
MMP-13 活性抑制分析 ---------------------------------------------------------- 40
4-5. 山竹、久司巴西酵素及大麥若葉水萃取液之 DPPH 自由基清除率分析
-------------------------------------------------------------------------------------------- 52
第五章 結論 ---------------------------------------------------------------------------- 56
參考文獻 -------------------------------------------------------------------------------- 58

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