In vitro 및 in vivo 에서 미꾸라지 근육 peptide의 항산화 효과 비교
Comparison of In vitro and In vivo Anti-oxidant Effects of Loach Muscle Peptides
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초록
본 연구에서는 미꾸라지 근육단백질 가수분해물의 항산화능력을 평가하고 생체 내에서도 그 효과가 있는지를 평가하기 위해 in vitro와 in vivo 시험계를 병행하여 시험을 수행하였다. In vitro 평가방법인 DPPH radical 소거능 시험에서처리방법(단순 가열, 알칼리 및 효소분해, 가열 및 효소분해,효소분해)을 달리한 여러 시료가 모두 농도 의존적인 항산화능을 보였다. 또 다른 in vitro 시험계인 superoxide radical 소거능에서는 알칼리 및 효소분해군에서만 소거능을 보였다. 한편 mouse에서 사염화탄소 투여로 야기한 간장독성 시험 model을 이용한 in vivo 시험계에서는 항산화효과를 확인할 수 없었다. 이 결과는 비록 in vitro 시험계에서 항산화 효과가 관찰된다고 하더라도 그 물질의 섭취 시생체내에서도 항산화 효과를 발휘할 지는 의문스러움을 시사한다. In vivo에서 항산화 효과가 나타나지 않는 원인으로서 활성물질의 위장관내에서의 파괴나 불량한 흡수와 같은요인을 고려해 볼 수 있을 것이다.
Abstract
The purpose of this study was to investigate anti-oxidant effects of loach muscle-derived peptides in vitro and in vivo. Loach muscle peptides were prepared by 4 different methods: boiling (B), enzymatic hydrolysis (E), boiling and enzymatic hydrolysis (BE), alkaline and enzymatic hydrolysis (AE). Two different in vitro analyses, DPPH radical scavenging activity and xanthine-xanthine oxidase-induced superoxide radical scavenging activity, were performed. All the four preparations showed concentration-dependent DPPH radical scavenging activity. However, superoxide radical scavenging activity was found only with AE and E preparations. To evaluate in vivo effects, mice were fed with 10% AE-containing diets for 4 weeks before hepatotoxicity induction with CCl4. In serum glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) levels, total antioxidant levels, and relative hepatic weight ratio, no evidence for anti-oxidant effects was found with AE indicating the absence of anti-oxidant effect in the in vivo mouse experiment. It needs to be clarified why anti-oxidant activity of loach protein hydrolysates was not evident in vivo. Furthermore, these results suggest that in vivo evaluation is crucial in demonstrating anti-oxidant activities.
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