臺灣博碩士論文加值系統

我們將本論文分做兩個部分進行探討。
第一部分：紫外線的輻射是造成許多皮膚病變的原因之一，長時間在紫外線的照射下會造成角質細胞的損傷。而紫外線輻射的中波UVB所造成細胞的損害尤其嚴重，可能造成DNA或蛋白質等分子的損害，當這些損傷過於嚴重的時候會使得角質細胞趨向於癌化。因此，如何抑制已損傷的角質細胞走向癌化並減緩皮膚癌的發生是本文研究的動機。先前國內外之研究證實穿心蓮內酯能夠發揮抗發炎及抗癌的效用，我們進一步探討穿心蓮內酯對於紫外線所造成的人類角質層細胞（HaCaT）的影響極其造成的死亡機轉。實驗中以不同濃度的穿心蓮內酯(0-50 μM)與UVB（1.5 mJ/cm2）的照射分別處理下，結果顯示在低劑量的穿心蓮內酯（1 μM ~ 5 μM）對HaCaT細胞是不具細胞毒性的，以及在UVB照射的試驗則發現當細胞單獨照射UVB 1.5 mJ/cm2的劑量會造成約4%的細胞死亡。而在穿心蓮內酯濃度(2.5 μM和5 μM)與1.5 mJ/cm2的UVB共同作用細胞後，可更加促進細胞走向凋亡。由此我們推測此傳統中草藥「穿心蓮內酯」具有預防紫外線照射後角質細胞癌化之潛力，對於抑制或減緩人類皮膚癌應有相當的裨益。
第二部分：活性氧分子（Reactive oxygen species; ROS）是造成氧化壓力的原因之ㄧ，氧化壓力的增加可能對細胞造成傷害，使細胞可能借由細胞凋亡或是細胞壞死的途徑而走向死亡。在近年來的研究也得知利用調控ROS的生成可抑制癌細胞的生長； 桂花（Osmanthus fragrans flower）含有許多的抗氧化物質，其萃取物具有增加人類肝癌細胞HepG2 cell氧化壓力的效果。在MTT及LDH的分析結果發現對HepG2具有生長抑制的效果。然而在本實驗中並未發現Osmanthus fragrans flower萃取物會造成HepG2凋亡的表現。此外，我們發現桂花萃取物也對於正常人類肝細胞Chang liver cell則有促進生長且增加細胞模的完整性的功能。因此，桂花萃取物可能應用在護肝上及肝癌相關的治療是具有相當潛力的。

We will devide it into two parts to introduction in the study.
In part one： Ultraviolet radiation（UV） is the cause of many skin diseases, and being exposing under ultraviolet radiation on long-time bases could cause the injury of keratinocyte cell. The UVB of Ultraviolet radiation could caused the most serious injury to cells, and the UVB could caused of injury in DNA or protein and lading to promoter tumor produced. Therefore, Is been interested by how to suppressed the cell injury. Function of andrographolide could to anti-cancer and anti- immunity. In our study, we used different concentrate of andrographolide ( 0 μM-50 μM )on effect of low dose UVB（ 1.5 mJ/cm2 ） induced apoptosis in human keratinocyte cell. In the result, low dose andrographolide （1 μM、2.5 μM、5 μM）is non toxic effect in HaCaT cell. Induced 4 % death by 1.5 mJ/cm2 of UVB in HaCaT cell, and the 2.5 and 5μM andrographolide combined UVB could enhance apoptosis landing to cell death. In future, can application use the andrographolide in inhibit UVB promoter tumor by Chinese medical herb in human keratinocyte cell, and reduce occurrence of skin cancer.
In part two：Oxidative stress is caused by exposure to reactive oxygen species, which can affect the damage effect of proteins and cell membranes, even lead to cell death via necrosis or apoptosis. So, the cancer cell inhibited by regulating intracellular ROS has been described in recently studies.
The extract of Osmanthus fragrans flower is rich with antioxidant compounds and enhances ROS produced in Hep G2 cell line. MTT and LDH assays showed that the extract of Osmanthus fragrans flower reduced cell viability in Hep G2 cell, but not showed apoptosis. Furthermore, no toxicity was found in the Chang liver cell treated with Osmanthus fragrans flower extract and must be maintain the integrity of the cell membrane. These results suggest that Osmanthus fragrans flower extract may be valuable for a hepatoprotective agent and possible to development of the therapeutic management of human hepatoma.

誌謝 iii
中文摘要 - 3 -
Abstract - 5 -
第一部分：穿心蓮內酯對紫外線輻射B誘導人類角質層細胞之影響
- 1 -
第一章 前言 - 2 -
一、常用中草藥植物 - 2 -
二、穿心蓮（ Andrographis paniculata ( Burm. F. ) Nees ） - 3 -
三、穿心蓮內酯 ( Andrographolide；Andro ) - 6 -
四、紫外線（Ultraviolet radiation ; UVR）簡介 - 9 -
五、細胞凋亡 - 11 -
六、研究目的 - 14 -
七、研究架構 - 15 -
第二章 材料與方法 - 16 -
第三章 結果 - 23 -
參考文獻 - 30 -
圖表 - 36 -
第二部分：桂花萃取物調控人類肝癌細胞株氧化壓力之影響 - 45 -
第一章 前言 - 46 -
一、氧化壓力 - 46 -
二、肝癌 - 47 -
三、天然藥草植物 - 49 -
四、桂花 - 50 -
五、目的 - 53 -
六、研究架構 - 54 -
第二章 材料與方法 - 55 -
第 四 章 結果 - 61 -
第五章 討論 - 63 -
圖表 - 69 -

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