황근 및 제주피막이 추출물의 기능성 화장품 소재 개발 연구

Abstract

In this study, weinvestigated anti-oxidative, anti-inflammatory and anti-bacterial constituents from Talipariti hamabo and Hydrocotyle japonica Makino. Five constituents were isolated from the extract of T. hamabo leaves; oleic acid (1), isoquercetin (2), nicotiflorine (3), 3-(4-hydroxyphenyl) propionicacid (4), p-hydroxyphenethyl-trans-ferulate (5). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including NMR spectra, as well as comparison of the data to the literature values. Upon the anti-oxidative studies with DPPH and ABTS+ radicals, potent radical scavenging activities were observed in the ethyl acetate (EtOAc) and n-Butanol (BuOH) fractions. T. hamabo leaves were in the anti-inflammatory tests using lipopolysaccharide (LPS)-stimulated RAW264.7 cells, the EtOAc fraction inhibited the nitric oxide (NO) production without causing cell toxicity, effectively. Moreover, the EtOAc fraction reduced pro-inflammatory cytokines tumor necrosis factor (TNF)-α, interleukin (IL)-6 and prostaglandin (PG) E2. Also, the EtOAc and n-BuOH fractions showed anti-bacterial activities against Staphylococcus epidermidis. Fifteen constituents were isolated from the extract of T. hamabo branches; methyl stearate（1)，oleic acid（2)，linolenic acid（3)，α-linolenic acid（4)，ethyl linolen ate（5)，friedelin（6)，friedelinol（7)，glutinol（8)，3β-olean-12-en-3-ol（9)，oleanolic-acid（10)，β-sitosterol（11)，p-hydroxyphenethyl-trans-ferulate（12)，caffeic acid（13), caffeic anhydride（14)，triumfettalarein (15). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including NMR spectra, as well as comparison of the data to the literature values. On the anti-oxidative studies with DPPH and ABTS+ radicals, potent radical scavenging activities were observed in the EtOAc fractions. Also, for the cellularprotective effects on HaCaT keratinocytes damaged by H2O2, compound 15 indicated protective effects against oxidative stress. T. hamabo branches were in the anti-inflammatory tests using LPS stimulated RAW264.7 cells, the n-hexane(Hex) and EtOAc fraction inhibited the NO production without causing cell toxicity, effectively. Moreover, the EtOAc fraction reduced pro-inflammatory cytokines TNF-α and compound 15 reduced pro-inflammatory cytokines TNF-α, IL-6，1L-1β and PGE2. Twenty constituents were isolated from the extract of H. japonica aerial parts; (E)-phytol (1)，neophytadiene（2)，ethyl oleate（3)，1-monoolein（4)，linolenic acid（5)，ethyl linoleate（6)，1-linoleoyl glycerol（7)，enthyl linolenate(8)，1-linolenoyl glycero（9)，1,9-octadecadiene-4,6-diyn-3S-ol（10 )，β -si t ost ero l（1 1 )，capsid iol-3 -acet at e（1 2 )，(-)-hi noki ni n（1 3 )，c h l o r o p h y l - 1 1 （1 4 ) ，( 2 S ) - 1 –O - ( 9 Z , 1 2 Z - o c t a d e c a d i e n o y l ) - 3 - O -β -galactopyr-anosylglycerol(15)，(2S)-1–O-(7Z,10Z,13Z-hexadecatrienoyl)-3-O-β-galacto-pyranosylglycerol(16)，isorhamnetin-3-O-β-D-glucopyranoside (17)，quercetin-3-O-β-D-galactoside（18), quercetin 3-O-6’’-caffeoyl-β-Dgalactopyran-oside（19)，ｚizyflavoside（20). Flavonoids compound 17, 18, 19, 20 in the anti-oxidative studies with DPPH and ABTS+ radicals potent radical scavenging activities were observed. In the anti-inflammatory tests using LPS stimulated RAW 264.7 cells, the extract, n-Hex，EtOAc and n-BuOH fraction inhibited the NO production without causing cell toxicity, effectively. Moreover, the extract and n-Hex fraction reduced pro-inflammatory cytokines TNF-α and 1L-1β. Compound 12,19 reduced pro-inflammatory cytokines TNF-α, IL-6, 1L-1β and PGE2. Compound 15 reduced pro-inflammatory cytokines TNF-α, IL-6 and PGE2. Compound 10 reduced pro-inflammatory cytokines TNF-α and IL-6. Also, the EtOAc fractions showed anti-bacterial activities against Staphylococcus epidermidis and Cutibacterium acnes. Based on these results, it was suggested that the extract and isolated compounds from T. hamabo and H. japonica could be potentially applicable as natural source for pharmaceutical and/or cosmetic ingredients.