Isolation and Identification of Constituents with Activity of Inhibiting Nitric Oxide Production in Raw 264.7 Macrophages from Gentiana triflora

Abstract

Gentiana triflora is widely used to treat inflammation, jaundice, hepatitis, and rheumatism. In this study, three new compounds, including a benzo seven-membered ring compound, gentioxepine (1), two secoiridoid glucosides, (1S,5R,9R)-deglucosyltrifloroside (2) and (1S,5R,9R)-scabraside (3), together with seven known ones, (+)-syringaresinol (4), deglucogelidoside (5), 3,4-dihydro-1H,6H,8H-naphtho[1,2-c:4,5-c',d']dipyrano-1,8-dione (6), deglucoscabraside (7), 2-hydroxy-3-O-β-D-glucosyloxy benzoic acid methyl ester (8), gentiolactone (9), and trifloroside (10), were isolated from the ethanol extract of Gentiana triflora. Their structures were mainly confirmed on the basis of NMR, MS, IR, CD, and UV spectral evidences. Inhibiting activities of nitric oxide production of eight of the compounds isolated, as well as gentiopicroside, were evaluated in the macrophage cell line RAW 264.7. The results show that the three new compounds and compound 7 could significantly suppress lipopolysaccharide-induced production of NO, with IC₅₀s of 2.2 µM, 37.5 µM, 17.6 µM and 6.9 µM, respectively. Among them, compounds 1, 3, and 7 showed stronger inhibitory activity than that of the clinically used drug indometacin. Other tested compounds exerted moderate inhibiting activities.

Key words

Gentiana triflora (Gentianaceae) - secoiridoid glucoside - nitric oxide production - RAW 264.7 macrophage cells

• References

• 1 Georgieva E, Handjieva N, Popov S, Evstatieva L. Comparative analysis of the volatiles from flowers and leaves of three Gentiana species. Biochem Syst Ecol 2005; 33: 938-947
• 2 Matsukawa K, Ogata M, Hikage T, Minami H, Shimotai Y, Saitoh Y, Yamashita T, Ouchi A, Tsutsumi R, Fujioka T, Tsutsumi KI. Antiproliferative activity of root extract from gentian plant (Gentiana triflora) on cultured and implanted tumor cells. Biosci Biotechnol Biochem 2006; 70: 1046-1048
• 3 Inouye H, Ueda S, Nakamura Y, Inoue K, Hayano T, Matsumur H. Trifloroside, new secoiridoid glucoside from Gentiana triflora var. japonica . Tetrahedron 1974; 30: 571-577
• 4 Jiang RW, Wong KL, Chan YM, Xu HX, But PP, Shaw PC. Isolation of iridoid and secoiridoid glycosides and comparative study on Radix Gentianae and related adulterants by HPLC analysis. Phytochemistry 2005; 66: 2674-2680
• 5 Li C, Wang MH. Anti-inflammatory effect of the water fraction from hawthorn fruit on LPS-stimulated RAW 264.7 cells. Nutr Res Pract 2011; 5: 101-106
• 6 Tamir S, Tannenbaum SR. The role of nitric oxide (NO center dot) in the carcinogenic process. Biochim Biophys Acta 1996; 1288: F31-F36
• 7 Hsu CL, Chang FR, Tseng PY, Chen YF, El-Shazly M, Du YC, Fang SC. Geranyl flavonoid derivatives from the fresh leaves of Artocarpus communis and their anti-inflammatory activity. Planta Med 2012; 78: 995-1001
• 8 Han Y, Jung HW, Lee JY, Kim JS, Kang SS, Kim YS, Park YK. 2,5-dihydroxyacetophenone isolated from Rehmanniae Radix Preparata inhibits inflammatory responses in lipopolysaccharide-stimulated RAW264.7 macrophages. J Med Food 2012; 15: 505-510
• 9 Murakami A. Chemoprevention with phytochemicals targeting inducible nitric oxide synthase. Forum Nutr 2009; 61: 193-203
• 10 Cao FH, Shao H, Li Q, Li JR, Li WQ, Li C. Anti-inflammatory activity of Gentiana striata Maxim. Nat Prod Res 2012; 26: 1038-1044
• 11 Lim H, Son KH, Chang HW, Kang SS, Kim HP. Inhibition of chronic skin inflammation by topical anti-inflammatory flavonoid preparation, Ato Formula (R). Arch Pharm Res 2006; 29: 503-507
• 12 Kwak WJ, Kim JH, Ryu KH, Cho YB, Jeon SD, Moon CK. Effects of gentianine on the production of pro-inflammatory cytokines in male Sprague-Dawley rats treated with lipopolysaccharide (LPS). Biol Pharm Bull 2005; 28: 750-753
• 13 Yu FR, Yu FH, Li RD, Wang R. Inhibitory effects of the Gentiana macrophylla (Gentianaceae) extract on rheumatoid arthritis of rats. J Ethnopharmacol 2004; 95: 77-81
• 14 Mathew A, Taranalli AD, Torgal SS. Evaluation of anti-inflammatory and wound healing activity of Gentiana lutea rhizome extracts in animals. Pharm Biol 2004; 42: 8-12
• 15 Chen L, Liu JC, Zhang XN, Guo YY, Xu ZH, Cao W, Sun XL, Sun WJ, Zhao MG. Down-regulation of NR2B receptors partially contributes to analgesic effects of Gentiopicroside in persistent inflammatory pain. Neuropharmacology 2008; 54: 1175-1181
• 16 Chen L, Wang HB, Sun XL, Sun WJ. Study on the analgesic and anti-inflammatory activities of Gentiopicroside. Tianran Chanwu Yanjiu Yu Kaifa 2008; 20: 903-906
• 17 Li YQ. The main pharmacodynamics research of gentiopicroside [dissertation]. Shanxi: Xibei University; 2002
• 18 Chen CX, Liu ZW, Sun ZR, Song CQ, Hu ZB. Studies on anti-inflammatory effect of gentiopicroside. Zhong Cao Yao 2003; 34: 49-51
• 19 Song QS, Gao KB, Fu KZ. Isolation and identification of gentiopicroside from the roots of Gentiana triflora Pall. Zhong Yao Tong Bao 1987; 12: 36-59
• 20 Holroyde JK, Orr AF, Thaller V. 3,7-Bis (hydroxymethyl)-1-benzoxepin-5 (2H)-one, a novel oxygen heterocyclic metabolite from cultures of the fungus Marasmiellus ramealis (Bull. ex Fr.) Singer. J Chem Soc [Perkin I] 1978; 1: 1490-1493
• 21 Ikeda T, Hutchinson CR, Meier H, Tietze LF. Stereochemical correlations of secoiridoid aglucones. Tetrahedron Lett 1984; 25: 2427-2430
• 22 Nie LY, Jin HZ, Yan L, Qin JJ, Zhang WD. Chemical constituents of Inula lineariifolia Turcz. Tianran Chanwu Yanjiu Yu Kaifa 2011; 23: 643-646
• 23 Ikeshiro Y, Mase I, Tomita Y. A Secoiridoid Glucoside from Gentiana scabra var. buergeri . Planta Med 1990; 56: 101-103
• 24 Kitanov GM, Spassov SL. A naphthodipyranodione from Gentiana asclepiadea . Phytochemistry 1992; 31: 1067-1068
• 25 Ikeshiro Y, Tomita Y. A New bitter secoiridoid glucoside from Gentiana scabra var. buergeri . Planta Med 1983; 48: 169-173
• 26 Kakuda R, Machida K, Yaoita Y, Kikuchi M. Studies on the constituents of Gentiana species. II. A new triterpenoid, and (S)-(+)- and (R)-(−)-gentiolactones from Gentiana lutea . Chem Pharm Bull 2003; 51: 885-887
• 27 Tan RX, Wolfender JL, Zhang LX, Fuzzati W, Marston A, Hostettmann K. Acyl secoiridoids and antifungal constituents from Gentiana macrophylla . Phytochemistry 1996; 42: 1305-1313