臺灣博碩士論文加值系統

穿心蓮(Andrographis paniculata Nees.)為爵床科植物，是一種民間傳統藥用植物，廣泛應用於各種用途，以往被用為抗炎、抗菌、解熱及免疫促進之傳統治療藥物。過多的活性氧自由基產生的氧化壓力，容易造成大腦損傷，目前已知氧化壓力易造成神經退化性疾病的產生。穿心蓮具有很好的抗氧化能力，因此本研究探討穿心蓮的神經保護功效。我們利用神經纖維母細胞株IMR-32細胞，經由過氧化氫與乙醇的氧化傷害模式，探討穿心蓮萃取物是否具有抗氧化的神經保護功效。利用過氧化氫及乙醇誘發細胞死亡的實驗模式下，加入過氧化氫及乙醇刺激24小時可誘導細胞死亡，加入不同濃度的穿心蓮萃取物(25、50、100、200 μg/ml )可保護細胞避免死亡。經實驗證實，該細胞死亡是經由細胞凋亡機制，穿心蓮可以減緩細胞凋亡的發生。綜合本研究的結果，中草藥穿心蓮萃取物具有保護神經細胞對抗氧化壓力的功效，或許未來可以應用於神經退化性疾病的預防或治療。

Andrographis paniculata Nees., Family Acanthaceae, is a traditional folk medicinal plants, widely used in various applications, and has been used as antiinflammatory, antibacterial, antipyretic and immune promoting traditional therapeutic drugs. Excessive reactive oxygen radicals produced by oxidative stress make the brain hurt easily. It has been known that oxidative stress could easily lead to the generation of neurodegenerative diseases and the extract of Andrographis paniculata is a good antioxidant. Therefore, we used the human neuroblastoma IMR-32 cells treated with hydrogen peroxide and ethanol oxidation to investigate whether Andrographis paniculata extract has antioxidant neuroprotective effects. Under the cellular experimental conditions, the use of hydrogen peroxide and ethanol can induce cell death. To study the protective effects of Andrographis paniculata extract, we treated cells with hydrogen peroxide/ethanol and various concentrations of Andrographis paniculata extract (25, 50, 100, 200 μg/ml) for 24 hours and found the extract can protect cells from death. We studied whether the cell death via apoptosis using ApoGlowTM assay kit and found that hydrogen peroxide/ethanol induces apoptosis and Andrographis paniculata extract decreases the adverse effects. Integrated our experimental results, Chinese herbal medicine Andrographis paniculata extract protects nerve cells against oxidative stress injury. Maybe it can be used for prevention or treatment of neurodegenerative diseases in the future.

口試委員審定書
授權書
目 錄 i
圖目錄 iv
致謝 v
縮寫表 vii
中文摘要 1
英文摘要 2
第一章 前言
1-1 氧化壓力 3
1-2 乙醇所造成氧化傷害 4
1-3 過氧化氫所造成之氧化傷害 5
1-4 神經系統的細胞 5
1-5 穿心蓮介紹
1-5.1穿心蓮的名稱 6
1-5.2穿心蓮的產地來源 7
1-5.3穿心蓮植物形態特徵 7
1-5.4 穿心蓮的功效 9
1-6 細胞凋亡 9
1-7 研究目的 10
第二章 材料與方法
2-1實驗設備 12
2-2 實驗材料 12
2-3 實驗方法
2-3.1 細胞株 13
2-3.2 IMR-32細胞培養 14
2-3.3 細胞繼代培養 14
2-3.4 冷凍細胞活化 15
2-3.5 細胞冷凍保存 15
2-3.6 細胞計數方法 16
2-3.7 穿心蓮萃取物 16
2-3.8 細胞凋亡 16
第三章 結果
3-1穿心蓮萃取物不造成IMR-32細胞毒性 18
3-2過氧化氫對IMR-32 細胞之氧化傷害 18
3-3乙醇對IMR-32細胞毒殺之影響 19
3-4過氧化氫和穿心蓮對IMR-32細胞存活率之影響 19
3-5 乙醇和穿心蓮對IMR-32細胞存活率之影響 20
3-6穿心蓮萃取物不造成IMR-32細胞凋亡 20
3-7過氧化氫對IMR-32 細胞氧化傷害之細胞凋亡 21
3-8乙醇對IMR-32細胞凋亡之影響 21
3-9 過氧化氫和穿心蓮對IMR-32細胞凋亡之影響 21
3-10 乙醇和穿心蓮對IMR-32細胞凋亡之影響 22
第四章 討論
4-1穿心蓮萃取物抑制IMR-32細胞凋亡 23
4-2穿心蓮萃取物對過氧化氫誘發細胞死亡之保護作用 23
4-3 穿心蓮萃取物對乙醇誘發細胞死亡之保護作用 24
4-4 結論 24
參考文獻 24
圖 31
作者簡歷 51

圖表目錄
圖一、H2O2對IMR-32細胞毒殺之影響 31
圖二、乙醇對IMR-32細胞毒殺之影響 33
圖三、穿心蓮對IMR-32細胞毒殺之影響 35
圖四、H2O2和穿心蓮對IMR-32細胞存活率之影響 37
圖五、乙醇和穿心蓮對IMR-32細胞存活率之影響 39
圖六、穿心蓮萃取物不造成IMR-32細胞凋亡 41
圖七、過氧化氫對IMR-32 細胞氧化傷害經由細胞凋亡 43
圖八、乙醇對IMR-32細胞凋亡之影響 45
圖九、穿心蓮逆轉過氧化氫對IMR-32細胞凋亡之影響 47
圖十、乙醇和穿心蓮對IMR-32細胞凋亡之影響 49

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