Tissue Distribution and Activity of Analogs as Anti-Human Hepatoma Factors in Hard Clam, Meretrix lusoria

• Main Authors: Cha-Chung Tu, 杜嘉純
• Other Authors: Zwe-Ling Kong
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/23046011168791227181

碩士 === 國立臺灣海洋大學 === 食品科學系 === 97 === The goal of this study was to explain the uncertained effect of hard clam hot water extraction and identify the distribution of bioactive compounds. We used ethyl acetate to extract steroids from alimentary tract and muscle in hard clam, which named as HCEAA and HCEAM. The cell viability is determined by MTT assay. The molecular mechanisms of apoptosis inducing by HCEAA was determined by DNA fragmentation, Annexin V-FITC/PI double stain, analysis of mitochondrial transmembrane potential and sub-G1 DNA distribution. The sterol compounds were analysis by TLC and HPLC. Determination of ROS generation by DCFH-DA stained. Evaluate the antioxidization activity by DPPH scavenging activity. The activity of epidioxysterols analogs compound of extract from hard clam, HCEAA and HCEAM, were significantly decreased Hep3B and HepG2 cell viability. HCEAA (Yield: 4.77%) have higher yield compared to HCEAM (Yield: 1.33%), and showed more strong effect on cell cytotoxicity by 2 folds. By using TLC and HPLC, epidioxysterol was presented in HCEAA and HCEAM. HCEAA can increase the intracellular ROS concentration. In vitro, it has DPPH scavenging activity. Therefore, this study suggested that, extract by ethyl acetate can get bioactive compounds and distribute in alimentary tract. But compared with WG30-50 (IC50 = 100 μg/mL), HCEAA’s cytotoxicity (IC50 = 397.95 μg/mL) is not outstanding. The bioactive compound of hard clam is not necessarily epidioxysterol.