| Summary: | 碩士 === 國立臺灣大學 === 動物科學技術學研究所 === 102 === Velvet antler (VA) is one of the most valuable Chinese traditional medicines and commercial functional foods with enhancing vital energy and immune system. In our previous studies, we had demonstrated the anti-inflammatory and immunomodulatory effects of cold water extraction of VA from Formosan samber deer (Rusa unicolor swinhoii). Thus, the purpose of this study is to identify the active components in the cold water extraction. The proximate composition of VA cold water extraction powder was 83.3% of crude protein and 10.4 % of ash. To further identify the active components in the extraction, the cold water extraction powder of VA was separated into three populations by ultra-filtration concentrator based on molecular weight: (1) >10 kDa; (2) 5-10 kDa and (3) <5 kDa, and evaluated their anti-inflammatory effects in-vitro using RAW 264.7 macrophage-like cells stimulated with lipoteichoic acid (LTA). The results of pro-inflammatory cytokines showed that the production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) by LTA stimulation were significantly suppressed after co-cultured with 5-10 kDa and < 5 kDa samples of VA extracts. Besides, heat treatment at 100°C for 15 minutes did not affect the anti-inflammatory effect of VA extract samples.
We further analyzed the protein profile by sodium dodecyl sulfate gel electrophoresis (SDS-PAGE) and liquid chromatography (LC)-Mass spectrophotometer (MS)/MS of the cold water extractions. Three possible proteins were identified as profilin-1 from bovine (Bos taurus), fatty acid-binding protein-4 from red deer (Cervus elaphus), and ubiquitin from dromedary (Camelus dromedarius). Additionally, we also used high-performance liquid chromatography (HPLC) to separate and purify the fractions from VA cold water extraction. We found that some fractions separated from VA cold water extraction powder still had ability to suppress the pro-inflammatory cytokines production. After further identify the fractions, we found most of them were the proteins or peptides from antler blood.
According to the findings, the active components in VA water extraction from Formosa sambar deer might be heat resistance and water soluble proteins or peptides under 10 kDa, and the proteins or peptides from antler blood. We will further compare the VA from Formosan samber deer with other species, to find the unique functional components.
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