나무수국 꽃의 iridoid 및 flavonol 배당체의 골격근세포 비대 유도 효능

Abstract

나무수국 (Hydrangea paniculata Siebold)은 범의귀과 (Saxifragaceae)의 낙엽 활엽 관목이다. Coumarin과 페놀성 성분들이 나무수국의 지상부에서 분리 보고 되었지만, 꽃에 대한 성분 연구는 수행된 적이 없다. 본 연구는 나무수국의 꽃에 대한 식물 화학적 연구를 수행하였다. 나무수국 꽃을 methanol로 추출하고, 용매분획을 실시하여 분획을 나눈 후, n-butanol 분획을 open column chromatography와 prep-HPLC를 진행하여 10종의 화합물을 분리 정제하여, NMR과 ESI-Mass 등의 분광학적인 data를 이용하여 6종의 iridoid (1-5, 10), 1종의 coumarin (6), 3종의 flavonol glycoside (7-9)를 확인하였다. 각 화합물은 kingiside (1), morroniside (2), sweroside (3), loganin (4), vogeloside (5), umbelliferone (6), quercetin-3-O-sambubioside (7), quercetin-3-O-neohesperidoside (8), kaempferol 3-O-sambubioside (9), secologanin dimethyl acetal (10)로 동정하였다. 또한, 나무수국 꽃 추출물과 분리 정제한 화합물의 골격근세포 (C2C12 myotube)에 처리하여, 골격근세포의 hypertrophy능을 평가한 결과, n-butanol 분획물과 물층에서 활성을 나타냈으며, loganin (4)과 kaempferol 3-O-sambubioside (9)가 활성물질임을 확인하였다. Loganin (4)과 kaempferol 3-O-sambubioside (9)은 10 nM의 농도에서 C2C12 myotube의 diameter를 각각 12.25%, 6.52% 증가시켰으며, myosin heavy chain (MHC)의 발현을 유도하였다. 또한 western blotting을 통하여 p70S6K, AKT의 phosphorylation을 유의미하게 증가시킴을 확인하였다. 나무수국 꽃에서 10종의 화합물을 분리 정제하여 구조를 동정하였고, loganin (4)과 kaempferol 3-O-sambubioside (9) 골격근세포의 분화와 비대를 촉진하는 물질로 확인하였다.

Keywords: Hydrangea paniculata, iridoid, flavonol glycoside, C2C12 myotube hypertrophy |Although there have been many studies on the above-ground part of Hydrangea paniculate Siebold, there have been no studies on flowers. In this study, I was curious about the compounds of hydrangea flowers and tried to isolated. Hydrangea paniculate Sieb. flowers were extracted with methanol and fractionated with n-hexane, ethyl acetate, n-butanol and water. Among them, the nbutanol fraction was subjected to open column and prep-HPLC to isolate ten compounds. The compounds identified using LC-MS and NMR were respectively kingiside (1), morroniside (2), sweroside (3), loganin (4), vogeloside (5), umbelliferone (6), quercetin-3-O-sambubioside. (7), quercetin-3-O-neohesperidoside (8), kaempferol 3-O-sambubioside (9) and secologanin dimethyl acetal (10). Loss of skeletal muscle mass occurs during aging (sarcopenia), disease (cachexia), or lethargy (contraction). However, a drug that can treat sarcopenia has not yet been released. Differentiation and hypertrophy of skeletal muscle cells play an important role. In this experiment, in order to find out the active ingredient of the Hydrangea paniculate Sieb. flowers, each compound was treated with 10 nM each in C2C12 cells, and the activity was confirmed in compounds 4 and 9 by confirming that the diameter of myotube and the expression of myosin heavy chain (MHC) were induced. did. The activation degree of p70S6K and AKT was confirmed with the two compounds at concentrations of 10 nM and 30 nM. In conclusion, ten compounds among the compounds of Hydrangea paniculate Sieb. flowers were isolated and their structures were confirmed. Among them, loganin (4) and kaempferol 3-Osambubioside (9) were predicted to promote differentiation and hypertrophy of skeletal muscle cells.