A fast and efficient method for purification of dihydromyricetin (3,5,7,3′,4′,5′-six hydroxy-2,3-dihydro flavonol; DMY) from Ampelopsis grossedentata was created by crystallization eight times at 25°C, and a purity of 98% was finally achieved. The purified DMY exhibited high oxygen radical absorbance capacity (ORAC) (30.21 μmol Trolox equiv/mg) and strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (half-maximal inhibitory concentration [IC50]=0.235 μg/mL). The addition of DMY could also effectively attenuate 2,2′-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced human erythrocyte hemolysis and cupric chloride (CuCl2)-induced human plasma lipid peroxidation via inhibition of intracellular reactive oxygen species (ROS) generation. It was also found that DMY (>12 μg/mL) treatment significantly inhibited intracellular malondialdehyde (MDA) formation. Meanwhile, DMY treatment significantly inhibited the obvious increase of anti-oxidant enzymes levels (superoxide dismutase [SOD]; glutathione peroxidase [GPX], and catalase [CAT]) induced by AAPH radicals, suggesting that stress defense mechanisms are associated with protection of DMY against intracellular oxidation.
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References
1.
MurakamiT, MiyakoshiM, ArahoD, MizutaniK, KambaraT, IkedaT, ChouWH, InukaiM, TakenakaA, IgarashiK. Hepatoprotective activity of tocha, the stems and leaves of Ampelopsis grossedentata, and ampelopsin. Biofactors, 2004; 21:175–178.
2.
TowatariK, YoshidaK, MoriN, ShimizuK, KondoR, SakaiK. Polyphenols from the heartwood of Cercidiphyllum japonicum and their effects on proliferation of mouse hair epithelial cells. Planta Med, 2002; 68:995–998.
3.
CheungLM, CheungPC, OoiVE. Antioxidant activity and total phenolics of edible mushroom extracts. Food Chem, 2003; 81:249–255.
4.
ZhangYS, NingZX, YangSZ, WuH. Antioxidation properties and mechanism of action of dihydromyricetin from Ampelopsis grossedentata. Acta Pharmaceutica Sinica, 2003; 38:241–244.
5.
ZhengQ, XuL, ZhuL, ChenJ, LiuF, ChenD, XuK, ChenM. Preliminary investigations of antioxidation of dihydromyricetin in polymers. B Mater Sci, 2010; 33:273–275.
6.
DuQ, CaiW, XiaM, ItoY. Purification of (+)-dihydromyricetin from leaves extract of Ampelopsis grossedentata using high-speed countercurrent chromatograph with scale-up triple columns. J Chromatogr A, 2002; 973:217–220.
7.
YoshikawaM, MurakamiT, UedaT, YoshizumiS, NinomiyaK, MurakamiN, MatsudaH, SaitoM, FujiiW, TanakaT. Bioactive constituents of Chinese natural medicines. III. Absolute stereostructures of new dihydroflavonols, hovenitins I, II, and III, isolated from hoveniae semen seu fructus, the seed and fruit of Hovenia dulcis THUNB.(Rhamnaceae): Inhibitory effect on alcohol-induced muscular relaxation and hepatoprotective activity. Yakugaku Zasshi, 1997; 117:108.
8.
RoschekBJr, FinkRC, McMichaelMD, LiD, AlberteRS. Elderberry flavonoids bind to and prevent H1N1 infection in vitro. Phytochemistry, 2009; 70:1255–1261.
9.
ZhouT, ZhouX. Isolation, structure determination and pharmacological activity of flavanonol from Ampelopsis grossedentata. Chin Pharm J, 1996; 31:458–460.
10.
GuoQ, YuanJ, ZengJ, HeX, LiD. Synthesis of dihydromyricetin–manganese (II) complex and interaction with DNA. J Mol Struct, 2012; 1027:64–69.
11.
LiuB, DuJ, ZengJ, ChenC, NiuS. Characterization and antioxidant activity of dihydromyricetin–lecithin complex. Eur Food Res Technol, 2009; 230:325–331.
12.
GaoJ, LiuB, NingZ, ZhaoR, ZhangA, WuQ. Characterization and antioxidant activity of flavonoid-rich extracts from leaves of Ampelopsis grossedentata. J. Food Biochem, 2009; 33:808–820.
13.
GuLJ, ZhaoMM, LiWZ, YouLJ, WangJF, WangHY, RenJY. Chemical and cellular antioxidant activity of two novel peptides designed based on glutathione structure. Food Chem Toxicol, 2012; 50:4085–4091.
14.
HuangD, OuB, Hampsch-WoodillM, FlanaganJA, PriorRL. High-throughput assay of oxygen radical absorbance capacity (ORAC) using a multichannel liquid handling system coupled with a microplate fluorescence reader in 96-well format. J Agr Food Chem, 2002; 50:4437–4444.
15.
WuS, WuJ, YinZ, ZhangJ, LongC, KennellyEJ, ZhengS. Bioactive and marker compounds from two edible dark-colored Myrciaria fruits and the synthesis of jaboticabin. J Agr Food Chem, 2013; 61:4035–4043.
16.
CheungLM, CheungPC, OoiVE. Antioxidant activity and total phenolics of edible mushroom extracts. Food Chem, 2003; 81:249–255.
17.
XinML, MaYJ, XuK, ChenMC. Structure-activity relationship for dihydromyricetin as a new natural antioxidant in polymer. J Appl Polym Sci, 2012; 128:1436–1442.
18.
PandeyKB, RizviSI. Resveratrol up-regulates the erythrocyte plasma membrane redox system and mitigates oxidation-induced alterations in erythrocytes during aging in humans. Rejuvenation Res, 2013; 16:232–240.
19.
EsterbauerH, Dieber-RothenederM, StrieglG, WaegG. Role of vitamin E in preventing the oxidation of low-density lipoprotein. Am J Clin Nutr, 1991; 53:314S–321S.
20.
HancockJT, DesikanR, NeillSJ. Role of reactive oxygen species in cell signalling pathways. Biochem Soc T, 2001; 29:345–349.
21.
HsiehYS, HsuCY. Oxidative stress and anti-oxidant enzyme activities in the trophocytes and fat cells of aueen honeybees (Apis mellifera). Rejuv Res, 2013; 16:295–303.
22.
RocheM, TarnusE, RondeauP, BourdonE. Effects of nutritional antioxidants on AAPH-or AGEs-induced oxidative stress in human SW872 liposarcoma cells. Cell Biol Toxicol, 2009; 25:635–644.
23.
SinhaR, El-BayoumyK. Apoptosis is a critical cellular event in cancer chemoprevention and chemotherapy by selenium compounds. Curr Cancer Drug Targets, 2004; 4:13–28.
24.
HoppsE, NotoD, CaimiG, AvernaMR. A novel component of the metabolic syndrome: The oxidative stress. Nutr Metab Cardiovasc Dis, 2010; 20:72–77.
25.
YuM, SatoH, SeikiM, ThompsonEW. Complex regulation of membrane-type matrix metalloproteinase expression and matrix metalloproteinase-2 activation by concanavalin A in MDA-MB-231 human breast cancer cells. Cancer Res, 1995; 55:3272–3277.
26.
MissirlisF, PhillipsJP, JäckleH. Cooperative action of antioxidant defense systems in Drosophila. Curr Biol, 2001; 11:1272–1277.
27.
YehC, HouM, TsaiS, LinS, HsiaoJ, HuangJ, WangL, WuS, HouLA, MaH. Superoxide anion radical, lipid peroxides and antioxidant status in the blood of patients with breast cancer. Clin Chim Acta, 2005; 361:104–111.
28.
BakanN, TaysiS, YilmazÖ, BakanE, KuşkayS, UzunN, GündoğduM. Glutathione peroxidase, glutathione reductase, Cu-Zn superoxide dismutase activities, glutathione, nitric oxide, and malondialdehyde concentrations in serum of patients with chronic lymphocytic leukemia. Clin Chim Acta, 2003; 338:143–149.
29.
PolatMF, TaysiS, GulM, CikmanO, YilmazI, BakanE, ErdoganF. Oxidant/antioxidant status in blood of patients with malignant breast tumour and benign breast disease. Cell Biochem Funct, 2002; 20:327–331.
30.
RenJY, ZhaoMM, WangHY, CuiC, YouLJ. Effects of supplementation with grass carp protein versus peptide on swimming endurance in mice. Nutrit, 2011; 27:789–795.
31.
ChenY, CowleyAW, ZouA. Increased H2O2 counteracts the vasodilator and natriuretic effects of superoxide dismutation by tempol in renal medulla. Am J Physiol Regul Integr Comp Physiol, 2003; 285:R827–R833.
32.
AhmadH, TianJ, WangJ, KhanMA, WangYX, ZhangLL, WangT. Effects of dietary sodium selenite and selenium yeast on antioxidant enzyme activities and oxidative stability of chicken breast meat. J Agric Food Chem, 2012; 60:7111–7120.
