臺灣原生種植物之抗癌活性研究

The Anti-tumor Effects of Taiwan Endemic Plants

游璧如(Pi-Ju Yu)

指導教授 ： 王靜瓊

臺北醫學大學/藥學院/生藥學研究所/碩士(2005年)

摘要

癌症一直為世界各國主要的死亡病因。在台灣，惡性腫瘤（malignant tumor）自民國71年起連續23年蟬聯國人主要死亡原因榜首。目前化學治療用藥副作用影響癌症病人的生活品質，且許多腫瘤對於藥物有抗藥性的產生。因此開發新的抗癌藥物一直是一項重要的工作。台灣的地理位置跨越熱帶及亞熱帶地區，擁有豐富的原生種植物資源。我們認為這些原生種植物是一個很好的開發新藥的來源。本論文第一部份選擇24種坪林山區台灣原生種植物，以70％丙酮萃取，利用MTT體外細胞毒性試驗測定各種植物萃取物對於胃癌細胞（AGS、SNU-1）之細胞毒性。結果顯示狹瓣八仙花（Hydrangea angustipetala Hayata）葉部萃取物對於腫瘤細胞生長抑制活性最顯著，且對於正常細胞的毒性較小。因此，我們以抗癌活性作為導向，經由管柱分析方法來分離狹瓣八仙花葉部70％丙酮萃取物。並利用P388D1細胞引起CDF1擔癌鼠之體內抗癌模式來評估狹瓣八仙花之抗腫瘤生長活性。發現40％甲醇沖提物（D40M）有很強的細胞毒性，對兩株胃癌細胞IC50皆小於5 μg/ml，且在10 mg/kg之劑量下可以有效延長擔癌鼠的存活天數（ILS：120％）。更進一步從D40M分離出兩活性成分：(+)-febrifugine與trans-3-p-coumaryalquinic acid。MTT試驗顯示兩者之細胞毒性皆有時間濃度依存性，且有誘導細胞走向程式性死亡之能力，而(+)-febrifugine活性較強，因此我們認為(+)-febrifugine可作為抗胃癌活性藥物開發的前導物，且值得進一步探討其作用機轉。 論文的第二部份則評估台灣原生種杜鵑花科杜鵑花屬植物：西施花、烏來杜鵑、金毛杜鵑、南澳杜鵑之莖部與葉部100％乙醇及50％乙醇萃取物，共16種樣品之抗癌活性。研究發現，南澳杜鵑葉部100％乙醇萃取物最具有開發抗癌藥物之活性，在P388D1 bearing CDF1體內試驗模式顯示，5 mg/kg劑量下可延長擔癌鼠之存活天數，值得進一步做研究。

關鍵字

胃癌 ； 狹瓣八仙花 ； 杜鵑花科 ； 細胞毒性 ； 抗癌活性

並列摘要

Taiwan is located in the subtropics and abounded with many endemic plants. These endemic plants of Taiwan are good sources for developing new drugs. Cancer is one of the serious causes of death in humans. However, many tumors have shown resistances to drug, and many chemotherapeutic drugs have also been reported to cause serious side effects in clinical studies. Therefore, it is urgent to develop new anti-tumor drugs. In this study, we evaluated 4 Ericaceae plants and 24 kinds of endemic plants of Taiwan. In the first part, 24 kinds of endemic plants were collected from mountains in northern Taiwan and extracted with 70％ Acetone. The cytotoxicity effects of these extracts were evaluated by MTT assay in human gastric cancer cell lines, AGS, SNU-1, and normal cell line, Chang Liver cells. The results indicated that Hydrangea angustipetala Hayata potential than other endemic plants in inhibiting the growth of tumor cells. Column chromatography combined with cytotoxic bioassay-guided fractionation was performed to isolate the cytotoxicity compounds in this study. The anti-tumor effect of H. angustipetala was also evaluated by P388D1 bearing CDF1 mice. The 40％ MeOH eluted fraction（D40M）by diaion gel showed better anti-tumor effects in both in vitro and in vivo studies. IC50 values of D40M were 3.47 μg/ml in AGS and 3.48 μg/ml in SNU-1 cells, and the percentage of life span increase was 120％ in P388D1 bearing CDF1 mice model. Moreover, (+)-febrifugine and trans-3-p-coumaryalquinic acid were isolated as active anti-tumor products in H. angustipetala for the first time. Both compounds were able to suppress the proliferation of AGS and SNU-1 cells in a time- and dose-dependent manner. Based on the results from the Western blotting assay, these products induced apoptosis in human gastric cancer cell lines. The (+)-febrifugine showed stronger cytotoxicity than trans-3-p-coumaryalquinic acid in both in vitro and in vivo studies. Taken together, we suggested that (+)-febrifugine from Hydrangea angustipetala Hayata was a good leader compound to develop an anticancer drug. In studying of the anti-tumor effects of Ericaceae plants, 16 samples from four kinds Ericaceae Rhododendron plants were examined by MTT assay. Among them, the 100％ EtOH extracts of R. kanehirai Wilson and R. breviperulatum Hayata showed stronger cytotoxicity, and were evaluated by P388D1 animal model. The result indicated that the 100％ EtOH extracts from the leaves of R. breviperulatum could prolong the survival days of CDF1 mice under 5 mg/kg, the percentage increase in life span was 134%.

並列關鍵字

gastric cancer ； Hydrangea angustipetala ； Ericaceae ； cytotoxicity ； anti-tumor effects

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