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

為因應臺灣高齡化及少子化衍生致伴侶動物市場成長及對伴侶動物保健產品之需求 。本研究利用水產加工副產物蝦頭萃取磷脂質製成微脂體(SH-Lip.)，分別以犬類巨 噬細胞株(DH82)、老鼠巨噬細胞株(RAW264.7)及犬初代培養之周邊血液單核球細胞 (canine-PBMC)進行免疫調節確校評估。分析SH-Lip.粒徑約129.2 nm，其外層膜電 位-31.7 mV；實驗以DH82共培養1 mg/mL以下的SH-Lip.對細胞活存率無顯著差異 ，在脂多醣與g-干擾素(LPS/IFN-g)共同誘導DH82發炎實驗中，添加0.037~1 mg/mL SH-Lip.顯著調升抗發炎基因TGF-b及IL-10的表現量(P<0.05)，添加0.11~1 mg/mL SH-Lip.顯著調降細胞上清液中促發炎細胞激素IL-1b、IL-6、IL-12及巨噬細胞趨化 蛋白MCP-1的分泌(P<0.05)；進一步在LPS誘導canine-PBMC發炎試驗中，添加0.11~1 mg/mL SH-Lip.亦顯著調降細胞激素IL-6、IL-10、IL-12及TNF-a的分泌(P<0.05)。 以流式細胞儀分析RAW264.7極化之細胞表面抗原CD86 (M1型)與CD206 (M2型)，結果 在LPS誘導後RAW264.7細胞極化M1/M2比例由0.41上升致0.77，若添加0.037~1 mg/mL SH-Lip.共培養，M1/M2 比例降為0.58~0.45，顯示共培養SH-Lip.有助於M1/M2的平 衡。上述結果初步顯示，SH-Lip.應具有調節DH82、canine-PBMC及RAW264.7之免疫 活性，未來應可應用於伴侶動物皮膚保健產品之開發。

The companion animal marker is growth to response the trend of fewer children and more elderly society in Taiwan, therefore the companion animal healthcare product development is expected. In this study, a liposome (SH-Lip.) was prepared by extracting phospholipids from shrimp head which is the by-products from seafood processing wastes. We were validated the immune regulation effects on canine macrophage cell line (DH82), mouse peritoneal macrophage cell line (RAW264.7) and the primary cell culture from canine peripheral blood mononuclear cell (canine-PBMC). The SH-Lip particle size was 129.2 nm approximately, and the potential on outer membrane was -31.7 mV. The cell viabilities were no significant affected, when co-cultured with below 1 mg/mL of SH-Lip. on LPS induced DH82 cell. On the lipopolysaccharide and interferon-g (LPS/IFN-g) costimulated DH82 cell inflammatory cell based assay, the anti-inflammatory gene expressions were up-regulated when the cell co-cultured with 0.037~1 mg/mL of SH-Lip., including TGF-b and IL-10. The pro-inflammatory cytokines such as IL-1b, IL-6, IL-12 and the monocyte chemoattractant protein-1 (MCP-1) in the supernatants were inversely significant downregulated, when the cell co-cultured with 0.11~1 mg/mL of SH-Lip. (P<0.05). Furthermore, the cytokines IL-6, IL-10, IL-12 and TNF-a secretions also markedly decreased (P<0.05) on LPS induced canine-PBMC cell, when the cell co-treated with 0.11~1 mg/mL of SH-Lip. On the other hands, the polarization of cell surface markers CD86 (M1) and CD206 (M2) were also analysis by flow cytometry. The results showed that M1/M2 ratio on the LPS induced RAW264.7 polarization is increase from 0.41 to 0.77, and the ratio is reduced to 0.58~0.45 when the cell co-cultured with 0.037~1 mg/mL of SH-Lip., seemed the co-culture SH-Lip help balance of M1/M2 ratio.Those results preliminarily showed that SH-Lip. should have the immunological activity for regulating DH82, canine-PBMC and RAW264.7 cells. SH-Lip. might be applied to the development of companion animal skin health products in the future.