洛神花青素抑制黑色素瘤細胞轉移之作用

Chemoinhibitory Effect of Hibiscus anthocyanins Extract on the Metastasis in B16 Melanoma Cells

林婉玉(Wan-Yu Lin)

指導教授 ： 張雲菁

中山醫學大學/醫學院/生化暨生物科技研究所/碩士(2010年)

摘要

洛神花，其名稱是由英名Roselle音譯而來，原產於印度等地，學名為Hibiscus sabdariffa Linnaeus，目前產地遍及熱帶及亞熱帶地區，在台灣東部以及中南部也有大量種植。洛神花主要有效成分包含原兒茶酸（protocatechuic acid，PCA）、花青素（anthocyanin）、類黃酮素（flavonoid）以及異黃酮（isoflavonoid）。根據許多研究指出，癌症轉移（cancer metastasis）是致死的主要原因之一，因轉移的癌細胞經由局部侵入（invasion）周遭正常組織，甚至經由體內循環及淋巴系統轉移到身體其他部位，大幅提升了治療上的困難度，因此，在無法防止癌症形成之際，若可試圖阻斷癌細胞轉移，將會是一個極具發展潛力的方向。本篇研究目的只在觀察分離自洛神花花青素（Hibiscus anthocyanin，HAs）是否能夠抑制腫瘤的轉移及與其密不可分的血管新生之作用。首先，利用細胞毒性實驗觀察HAs對老鼠黑色素瘤細胞（B16-F1 cell）的影響，進而選用適當劑量（0~3 mg/ml）的花青素進行雞胚胎實驗，證實HAs能抑制血管新生作用，並且隨著濃度的增加，其抑制活性亦加明顯。進一步以動物模式餵食HAs來探討其對黑色素瘤細胞轉移影響，發現餵食HAs其小黑鼠體內腫瘤轉移現象有明顯抑制，且腫瘤生長情形亦顯著受到抑制。綜合以上結果，推測HAs具有降低腫瘤轉移及抗血管新生作用的潛力，未來希望本研究的初步成果可應用在日後保健食品之開發，使人人的健康保健觀念可更為提升。

關鍵字

黑色素瘤 ； 轉移 ； 花青素 ； 洛神花 ； 血管新生

並列摘要

The character of Roselle is transliterated from English alphabet. Hibiscus sabdariffa Linnaeus or Roselle is a member of Malvaceae family. Roselle is an attractive plant believed to have originated from India, and is widely grown in the tropic, also cultivated in the Sudan and Eastern Taiwan. The main active ingredient in Roselle contained protocatechuic acid, anthocyanins, flavonoids and isoflavones. Cancer metastasis is one of the lethal causes in the late stages of cancer. Many papers indicate that metastasis of cancer cells often invade surrounding normal tissues by local or whole body circulation and lymphatic system to increase the difficulty of treatment. We could not stop the development of cancer; however, trying to block or delay tumor metastasis would be a potential direction of development. This study is observe the effect of anthocyanins isolated from roselle (Hibiscus anthocyanin, HAs) to inhibit tumor metastasis and angiogenesis. First, the use of cell toxicity tests to observe that HAs inhibit B16-F1 cells growth or sensitivity in vitro. From the MTT data, we choice appropriate concentration (0-3 mg/ml) to process CAM analysis. The experiments are demonstrated that HAs could inhibit angiogenesis with the increasing dose, the better inhibition. Additionally, data showed that the inhibition of tumor metastases and tumor growth induced by HAs treatment in a C57BL/6 mice model. The results implied anti-metastatic effect of HAs display in vivo. Takntogether, we suggested that the HAs can promote suppression of tumor metastasis and angiogenesis. Hopefully the preliminary results in this study could be applied in the future health food development.

並列關鍵字

Hibiscus sabdariffa Linnaeus ； Anthocyanin ； Metastasis ； Angiogenesis ； Melanoma

參考文獻

1.王朝鐘，植物紅寶石洛神花之醫學功能與產品開發。食品生技。NO.14,70-7,2008。

2.Chen CC, Chou FP, Ho YC, Lin WL, Wang CP, Hao ES, Huang AC, Wang CJ.(2004). Inhibitory effects of Hibiscus Sabdarriffa L. extract on low-density lipoprotein oxidation and anti-hyperlipidemia in fructose-fed and cholesterol-fed rats. J Sci Food Agric. 84, 1989-1996.

3.Wang CJ, Wang JM, Lin WL, Chu CY, Chou FP, Tseng TH. (2000). Protective effect of Hibiscus anthocyanins against tert-butylhydroperoxide-induced hepatic toxicity in rats. Fd Chem Toxicol. 38, 411-416.

4.Lee MJ, Chou FP, Tseng TH, Hsieh MH, Lin MC, Wang CJ.(2002). Hibiscus protocatechuic acid or esculetin can inhibit oxidative LDL induced by either copper ion or nitric oxide donor. J Agric Fd Chem. 50, 2130-2136.

5.Chang YC, Huang KX, Huang AC, Ho YC, Wang CJ.(2006). Hibiscus anthocyanins-rich extracts inhibited LDL oxidation and oxLDL-mediated macrophages apoptosis. Food Chem Toxicol. 44, 1015-1023.