臺灣博碩士論文加值系統

紫珠(Callicarpa sp.)又稱beautyberry，主要在亞洲的熱帶和亞熱帶地區的藥用植物，在先前研究中指出在中國醫療上用以止血化瘀、治療肝炎、風濕病、發燒、頭痛、消化不良等疾病，紫珠目前的研究發現具有抗菌、抗氧化、抗發炎、抗腫瘤等活性，但對前列腺癌方面的研究較少，且其機制目前也都尚未釐清，因此本篇利用12種不同萃取條件之紫珠葉萃取物進行生物活性的探討。本研究藉由MTT試驗進行前列腺癌細胞(DU-145、PC-3)存活測試、Gelatin Zymography分析轉移蛋白MMP-2蛋白的活性以及DPPH、總多酚與DCFH-DA進行抗氧化能力分析。MTT試驗結果顯示紫珠葉乙醇萃取對於前列腺癌細胞增生皆有顯著抑制現象；腫瘤轉移試驗中發現，不同紫珠葉萃取樣品在最低濃度皆能抑制MMP-2蛋白的活性。抗氧化的評估試驗發現，紫珠葉在95 % 乙醇室溫搖晃之萃取濃縮下有最好的DPPH清除自由基的能力；在70 %乙醇室溫搖晃萃取之濃縮下可以萃取出比較多的多酚化合物；在95 % 乙醇室溫搖晃之萃取濃縮下，可以萃取出較多的類黃酮化合物，DCFH-DA不同條件萃取條件之萃取物在最低濃度62.5 μg/ml即有明顯抑制H2O2誘導的氧化傷害且具有最佳的細胞抗氧化能力。綜合以上結果，紫珠葉萃取物具有抑制腫瘤增生、抑制腫瘤細胞轉移、抗氧化等生物活性。

Callicarpa sp., also known as beautyberry, scientific name Callicarpa sp. is a medicinal plant mainly used in tropical and subtropical regions of Asia. The current research of Callicarp sp. has found that it has antibacterial, antioxidant and anti-inflammatory activity. Although anti-tumor activity has been studied such as the inhibition effect on breast carcinoma cell, the effect of Callicarpa sp. on prostate cancer is still unclear. Therefore, the leaves of Callicarpa sp. have been extracted by twelve different methods such as different solvents, temperatures and w/ or w/o sonication etc. and the bioactivity of leaf extracts from Callicarpa sp. would be studied in this study. According to our results, the leaf extracts from Callicarpa sp can significantly inhibit the growth or prostat cancer cells (DU-145 and PC-3) determined by MTT assay, and also can inhibit the expression of MMP-2 protein on DU-145 cell evaluated by gelatin zymography analysis. Moreover, the leaf extracts from Callicarpa sp. had antioxidant capacity demonstrated by polyphenol and flavonoid content. DCFH-DA extracts under different extraction conditions at the lowest concentration of 62.5 μg/ml can significantly inhibit the oxidative damage induced by H2O2 and have the best cellular anti-oxidation activities. DPPH and DCFH-DA assay. Based on the results, the leaf extracts from Callicarpa sp. have several bio-activities such as the inhibition of tumor cell growth, anti-metastasis and anti-oxidation activities.

第一章 文獻回顧 III
1.1紫珠(Callicarpa sp.)1
1.1.1紫珠介紹及分類1
1.1.2紫珠的化學組成2
1.1.2.1萜類(Terpenoids)2
1.1.2.2黃酮類(Flavonoids)2
1.1.2.3苯乙醇苷類(Phenylethanoid glycosides)3
1.1.2.4苯丙素類(Phenylpropanoids)3
1.1.3紫珠的藥理活性3
1.1.3.1 抑菌活性3
1.1.3.3 抗發炎4
1.1.3.4 抑癌活性4
1.2超音波萃取5
1.2.1超音波萃取原理5
1.2.2超音波萃取應用5
1.3.1前列腺癌介紹6
1.3.2 前列腺癌的治療7
1.3.3癌細胞轉移8
1.4.1 自由基8
第二章 研究動機10
第三章 實驗材料與方法11
3.1紫珠葉來源與樣品製備11
3.1.1實驗樣品來源11
3.1.2樣品製備11
3.1.2.1熱水萃取11
3.1.2.2超音波熱水萃取11
3.1.2.3乙醇萃取12
3.1.2.4超音波乙醇萃取13
3.2 細胞株14
3.3 實驗架構14
3.4抗氧化能力試驗15
3.4.1總多酚含量試驗( Total polyphenol content)–化學法15
3.4.2清除自由基DPPH–化學法15
3.4.3 總類黃酮 –化學法16
3.5細胞培養16
3.6 細胞存活率試驗( MTT assay)16
3.7 Gelatin Zymography Assay18
3.7.1 細胞培養液(conditioned medium)及細胞溶解液(cell lysate)的收集18
3.7.2 蛋白質含量測定18
3.7.3 膠體電泳分析(SDS -PAGE)18
3.7.4 西方墨點法(Western blot)19
第四章 實驗結果20
4.1 抑癌試驗20
4.1.1細胞存活測試20
4.1.2正常細胞生長試驗24
4.2 轉移酵素測試25
4.2.1 對DU-145分泌的基質金屬蛋白酶MMP-2的表現活性25
4.3 抗氧化能力測定27
4.3.1 DPPH抗氧化能力27
4.3.2 總多酚含量檢測27
4.3.4 對CHO-K1細胞的抗氧化能力試驗28
第五章 討論30
5.1抑制癌細胞生長及正常細胞毒性試驗30
5.2 基質金屬蛋白酶活性試驗30
5.3抗氧化試驗31
第六章 結論32
第七章 未來方向33
參考文獻34

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