고수온에 의한 틸라피아 HSP70 유전자 발현과 Quercetin의 발현 억제 효과
Inhibitory Effect of Quercetin on the Expression of HSP70 Gene Induced by High Water Temperature in Tilapia
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초록
수온은 어류를 포함한 수중동물의 다양한 생물학적 사건에 영향을 미친다. 어류의 번식 및 발생도 수온의 변화와밀접한 관련이 있다. 어류를 높은 수온에 노출시키면 HSP70 유전자 발현을 유도한다. 따라서, HSP70 유전자 발현을 효과적으로 차단할 수 있는 저해제의 개발은 온도에 민감한 생물학적 사건과 관련된 HSP70의 역할을 연구하는데 크게 기여할수 있다. 본 연구에서는 고온(36℃)에 노출된 틸라피아 자어와 치어(전장 10～13 ㎝)에서 천연 flavonoid의 일종인 quercetin,3,3',4',5,7-pentahydroxyflavon의 “고온 유도 HSP70 유전자 발현” 억제효과를 조사하였다. 고온 노출 전에 50 μM 이상의quercetin 용액에 6시간 동안 사전 침지한 실험어에서는 HSP70 유전자 발현이 유의하게 감소하였다(P<0.05). 특히, 100μM로 침지 처리한 개체에서는 고온 처리를 하지 않은 실험어에서 만큼 낮은 HSP70 유전자 발현을 나타내었다. 한편,0.1 ㎖씩의 0.5 mM, 5 mM 또는 20 mM의 quercetin 용액을 치어에 주사한 결과, 고수온 유도 HSP70 유전자 발현은 0.5mM의 quercetin을 주사한 치어의 생식소와 뇌에서 모두 유의하게 감소되었다(P<0.05). 본 연구의 결과는 quercetin이 어류에서 고온에 의해 유도된 HSP70 유전자 발현을 효과적으로 차단할 수 있으며, 틸라피아 자어 침지의 경우 유효 농도 100 μM 그리고 치어 주사의 경우 유효 농도 0.5mM로 원하는 효과를 얻을 수 있음을 제시한다.
Abstract
Water temperature governs various biological events in many aquatic animals including fish. Temperature changes the rates of gametogenesis and development, in some cases, is even capable of reversing fish sex. Treatments of fish with unusually high temperature are known to induce the expression of HSP70 gene. Development of an effective inhibitor for HSP70 gene expression is, thus, crucial to study the role of HSP70 in the temperature sensitive biological events. We have investigated the inhibitory effect of quercetin, 3,3',4',5,7-pentahydroxyflavon, a natural flavonid, on the expressions of HSP70 gene induced by high temperature (36℃) in the Nile tilapia, Oreochromis niloticus, larvae and juveniles (10～13 ㎝ in total length). The expression of HSP 70 gene was significantly decreased in tilapia larvae immersed in 50 μM or 100 μM quercetin solution for 6 hours before the exposure to high temperature (P<0.05). In particular, the level of HSP70 expression in fish treated with 100 μM was as low as that of fish without high temperature treatment. Juveniles of tilapia were individually injected with 0.1 ㎖ of either 0.5 mM, 5 mM or 20 mM of quercetin solution before the exposure to high temperature. As the results, the expression of HSP70 gene in the gonad and brain of juvenile fish was significantly inhibited by the injection of 0.5 mM quercetin solution (P<0.05), but not by higher concentrations. We report, for the first time in the fish, that quercetin effectively inhibits the expression of HSP70 gene induced by high temperature and 100 μM for the immersion of larvae and 0.5 mM for the injection to juvenile can be used for the effective concentrations for the study of temperature sensitive biological events in tilapia.
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