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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/15993

Title: 不同孵化時間點帶石斑魚受精卵萃取物對皮膚纖維母細胞之增生作用
Cell Proliferation Activities of Fertilized Roe Extracts of Epinephelus coioides at Different Hatching Stages on Skin Fibroblasts
Authors: Shih-Ping Jhang
張詩萍
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: Epinephelus coioides;魚苗萃取液;人類皮膚纖維母細胞;細胞增生
Epinephelus coioides;Larvae Extract;human skin fibroblast;cell proliferation
Date: 2010
Issue Date: 2011-06-30T08:27:25Z
Abstract: 海水魚類中石斑魚是一種具有相當高經濟價值的養殖魚種。本研究以石斑魚苗不同孵化階段,探討其抗皮膚老化之功能,並建立最適生產條件。以三倍體積的磷酸緩衝溶液 (滲透壓:280 mOs/kg 和 pH7.45) 在溫度 4oC 下均質不同孵化階段石斑魚苗,並且以 27,400 xg 離心去除多餘組織收集上清液,得到不同孵化階段為 ECLE 8 h (受精卵)、ECLE 16 h (受精卵剛孵化)、ECLE 40 h (魚苗孵化一天後)、ECLE 56 h (魚苗孵化兩天後)及 ECLE 80 h (魚苗卵黃囊消失並張口)總共五個階段水萃取液,且水萃液利用 0.22 m 過濾膜過濾以防止微生物生長。分析水萃液中蛋白質、還原醣濃度及水分含量,發現蛋白質濃度會隨著孵化時間增加而隨之減少,而還原醣濃度及水分含量無顯著變化。利用 MTT assay 測試對人類皮膚纖維母細胞及人類皮膚角質細胞增生效果,以 Collagen kit 測定萃取液對人類皮膚纖維母細胞其膠原蛋白胞外分泌量影響,以 RT-PCR 觀察萃取液對人類皮膚纖維母細胞膠原蛋白胞內基因表現量影響,以 SDS-PAGE 觀察萃取液蛋白質分子量分佈情形,以清除 DPPH 自由基能力來初步觀察萃取液是否有抗氧化功效。實驗顯示,ECLE 40 h 可穩定促進人類皮膚纖維母細胞增生至 202%,增加為 2 倍之膠原蛋白分泌量,增加胞內膠原蛋白基因表現 1.2 倍,且具有清除 DPPH 自由基能力達 42%。離心式過濾器分離不同分子量蛋白質,各分液初步觀察對於皮膚纖維母細胞增生影響,發現石斑魚苗萃取液在蛋白質分子量 30-100 kDa 均可使人類皮膚纖維母細胞增生至 150%。綜合以上結果,ECLE 40 h (魚苗孵化一天後)為最好採樣階段。
Aging comprises various changes that occur in living organisms with the passage of time, leading to increased system entropy, loss of homeostasis and eventually death. This study extracted larvae from five different Epinephelus coioides hatching stages of zygote, hatching, a day after hatching, two days after hatching, and yolk was absorbed (fertilization after 8, 16, 40, 64, and 80 hours) all by 3x volume of cold phosphate phosphatebuffered saline (osmotic pressure: 280 mOs/kg, pH 7.45), then homogenized under 4 oC and centrifuged at 27,400 xg. The supernatant was than filtered with 0.22 m micropore film to eliminate microorganisms. The protein extracted from Epinephelus coioides at the five different stages of hatching using Bradford protein-binding assay. The concentrations of protein decreased with increase hatching days. ECLE 40 h increased cell viability of CCD-966SK to 202% by MTT assay, and increased cell viability of HaCaT to 152%. ECLEs 40 h increased cell collagen secretion increased by 2.0 folds and enhancing collagen gene expression by 1.2 folds. 30-100 kDa in ECLE increased cell viability of CCD-966SK to 150% by MTT assay. ECLE 40 h demonstrated the DPPH radical scavenging activity. Taken together, these results suggest that ECLE 40 h (a day after hatching) was the best stage.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M97320041
http://ntour.ntou.edu.tw/ir/handle/987654321/15993
Appears in Collections:[食品科學系] 博碩士論文

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