本論文中使用之NML-E-E01是萃取自傳統中草藥植物廣藿香(Pogostemon cablin; PC)的沸水萃取物,傳統上廣藿香是用來治療感冒、反胃、腹瀉、頭痛及發燒等症狀。本研究中探討它的水萃取物對於人類神經母細胞瘤的抗腫瘤生長與老鼠巨噬細胞RAW264.7抗發炎的功效。 本研究利用體外實驗來探討NML-E-E01的抗腫瘤與抗發炎功效,分別利用SHSY-5Y細胞及RAW264.7細胞來進行分析。其中SHSY-5Y細胞與RAW264.7細胞分別給予不同濃度之NML-E-E01 24至72小時後,加入MTT染劑4小時後,再以DMSO溶出紫紅色結晶,並用分光光度計以波長540nm進行分析。此外利用propidium iodide (PI)染劑與流式細胞儀來分析細胞週期,同時以PI及Annexin V兩種染劑來確定細胞是否進行細胞凋亡的研究。細胞凋亡以及DNA的含量利用WinMDI 2.8軟體來進行分析。 本研究同時觀察NML-E-E01可誘發SHSY-5Y細胞的細胞凋亡與減少由LPS誘發RAW264.7細胞中的一氧化氮生成。由以上結果可以得知,NML-E-E01具有抗發炎與抗腫瘤的功效,本研究的結果可推論廣藿香的藥理作用在於抗腫瘤與抗發炎上的應用將非常重要。
NML-E-E01 was extracted from Pogostemon cablin (PC) which is a herbal medicine traditionally applied to treat not only common cold, nausea and diarrhea but also headache and fever. In the study, we have investigated the anti-tumour and anti-Inflammatory activities of the NML-E-E01 for human Neuroblastoma and Raw 264.7 respectively . The aim of this study was to investigate the anti-tumor and anti-inflammatory properties of standardized PC water extract (NML-E-E01) in vitro. The SHSY-5Y cells and Raw 264.7 cells were treated with NML-E-E01 for 24 to 72 hours respectively. MTT dye (1 mg/mL) was added to each well for at least 4 h of treatment. The cells (SHSY-5Y) were treated with NML-E-E01 for 24 hours. One milliliter of freshly prepared propidium iodide (PI) DNA staining solution was used to resuspend the cells. Stained cells were analyzed by flow cytometric analysis. Apoptosis and DNA content analysis by flow cytometry analysis on a FACS Calibur system (BD, USA). Data was analyzed by WinMDI 2.8 software (BD, USA). Effects of NML-E-E01 on human Neuroblastoma cancer lines (SHSY-5Y) were investigated in this study. In addition to the activities of inhibiting cell proliferation and increasing apoptosis, other mechanisms have also been proposed to rationalize the anticarcinogenic effect of NML-E-E01, such as the anti-inflammatory and antioxidant activities and others. Interest in this agent has grown in recent years based on its putative beneficial pharmacological effects, which include antioxidant, anti-inflammatory, and antitumor properties. These findings suggest that PC have produced an inhibition of cell proliferation as well as the activation of apoptosis in cancer therapy