Evaluation on Antioxidant and Antitumor of Ethanol Extracts from Seahorse Hippocampus kuda

• Main Authors: Ou, Chein-Yu, 歐倩妤
• Other Authors: Hwang, Deng-Fwu
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/dpmz96

碩士 === 國立臺灣海洋大學 === 食品科學系 === 105 === Seahorse is a small marine teleost animal. It has been used as traditional medicine for thousands of years in Eastern Asia because it has positive effects on human’s health. In this study, seahorses were collected from Penghu, freezing-dried and grounded into powder. The biochemical compositions of Hippocampus kuda (HK) powder were analyzed as follows: crude protein 69.16% (DW), crude lipid 0.86%, ash 28.06%, total saccharide 1.00 mg/g, total phenolic content 0.31 mg/g, and total flavonoids 0.03 mg/g. Furthermore, seahorses were derived into three parts. Two parts were extracted with 15% and 70% ethanol, and freezing-dried as 15HK and 70HK extracts. One part was extracted by 15% ethanol and further hydrolyzed by pronase E. The hydrolysate was as 15PHK hydrolysate. The protein content was 29.33, 12.13 and 7.97 mg/g DW in terms of 15HK extract, 15PHK hydrolysate and 70HKextract, respectively, and respective total saccharide content was 13.60, 11.75 and 31.53 mg/g, respective total phenolic content was 9.53, 13.28 and 11.87 mg/g, respective total flavonoid content was ND, ND and 2.43 mg/g. Three groups were also tested for their antioxidant activities including 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, reducing power and 2,2'-azino-bis(3-ethylbenzothiazoline -6 -sul- phonic acid (ABTS) radical scavenging activity The results showed that 15PHK has the potential ability of antioxidation. The results of antioxidant activities were 43.57%, 0.12 Abs. and 25.56%, respectively. 15PHK was purified with Hiprep DEAE FF 16/10 column by fast protein liquid chromatography (FPLC). Five fractions were abtained and tested their antioxidant activities. The results showed that 5 purified fractions of 15PHK appeared less antioxidant activity. Antitumor cell experiments were performed to incubate human cervix cancer cell line HeLa, human neuroblastoma cell line SH-SY5Y, human malignant melanoma cell line A375.S2, and human lung adenocarcinoma epithelial cell line A549 with 15HK, 15PHK and 70HK treatment. In MTT assay, A549 cell could be inhibited cell viability obviously by 70HK. A549 cell shrank after 24 hr 70HK incubation by observing. Moreover, lactate dehydrogenase (LDH) was released, nitric oxide (NO) was formed, and reactive oxygen (ROS) was measured corresponding with the more amount of 70HK. In contrast, mitochondrion membrane potential (MMP) was decreased. It showed that A549 cell incubated with 70HK treatment happened to apoptosis, and the more amount of 70HK treated, the more necrosis happened.