農業部-水產試驗所全球資訊網農業部-水產試驗所全球資訊網

本研究將麒麟菜 (Eucheuma serra) 經酵素萃取後所得之酵素水解物，進行探討酵 素水解物對 RAW264.7 類噬骨細胞之細胞增生與分化作用之影響。結果顯示，不同 濃度麒麟菜酵素水解物對 RAW264.7 巨噬細胞以及分化後 RAW264.7 類噬骨細胞 (添加分化劑 RANKL 和 M-CSF)，皆無細胞毒殺作用，且無促進細胞增生作用，而測 試促發炎 (IL-6 和 IL-1β) 和抗發炎 (IL-10) 等細胞激素部分則顯示麒麟菜酵素 水解物可降低促發炎細胞激素 IL-1β 至 46.6-79.5%，表示麒麟菜酵素水解物具有 抑制促發炎細胞激素 IL-1β 之作用。進一步用細胞染色鏡檢計數觀察發現，5-50 μg/mL 麒麟菜酵素水解物可抑制類噬骨細胞形成多核，並有劑量依賴性。另測試降 鈣素受體 Calcitonin receptor (CTR)，酵素水解物濃度為 ≧ 5 μg/mL 時，CTR 降至 53-73% (和正控制組 100% 相比)，皆顯著降低噬骨細胞表面降鈣素受體。在 評估骨質吸收作用方面，水解物濃度為 ≧ 31.25 μg/mL 時，有顯著差異，表示麒 麟菜酵素水解物可抑制噬骨細胞生成。機轉部分為觀察噬骨細胞內的 NFATc1、RANK 和 DC-STAMP 三種 mRNA 表現量以及一種 BCL-2 凋亡基因 mRNA 表現量之影響，結 果顯示 NFATc1 和 RANK 二種的 mRNA 表現量在酵素水解物濃度為 250-500 μg/mL 時，表現量有顯著差異 (和正控制組相比)，而 DC-STAMP mRNA 表現量則是當酵素 水解物濃度為 5-500 μg/mL 時，皆有顯著差異。當麒麟菜酵素水解物為 50-500 μg/mL 時，BCL-2 抗凋亡基因和 BAX 促凋亡基因 mRNA 表現量有顯著差異。綜觀 以上，推測麒麟菜酵素水解物對類噬骨細胞會造成細胞凋亡現象，進而抑制類噬骨 細胞分化及增生的能力，未來具有可應用為保骨素材發展之潛力。

The study is to investigate the bone health effect from E. serra by osteoclast-like RAW264.7 cells. Results showed that no cytotoxicity and proliferation in RAW264.7 and osteoclast-like RAW264.7 cells (added RANKL and M-CSF). The activity of pro-inflammatory factor (IL-6 and IL-1β) and anti-inflammatory factor (IL-10) assay showed that enzyme hydrolyzate can inhibited IL-1β activity to 46.6-79.5% from E. serra. 5-50 μg/mL enzyme hydrolyzate inhibited the formation of multinuclei in osteoclast-like RAW264.7 cells with a dose-dependent manner. The expression of calcitonin receptor were decreased to 53-73% (compared with positive control) with enzyme hydrolysate ≧ 5 mg/mL from E. serra. When enzyme hydrolysate ≧ 31.25 μg/mL, there were indicated that enzyme hydrolysate inhibit the production of osteoclasts in bone resorption assay. The mRNA expression of NFATc1、RANK and DC-STAMP were showed significantly different (compared with positive control) with 250-500 μg/mL enzyme hydrolysate. 50-500 μg/mL enzyme hydrolysate were showed significantly different in the antiapoptotic gene mRNA expression of BCL-2 and proapoptotic gene mRNA expression of BAX. In view of the above, enzyme hydrolysate can cause osteoclast cells apoptosis, and can inhibited osteoclast activity, with the potential of bone health from E. serra .