Dihydromyricetin (DHM) is a natural flavonoid showing several health promoting effects such as protective activity during severe alcohol intoxication. The mechanism underlying the effects of DHM on alcohol metabolism is virtually unknown. The present paper is focused on clarifying the role of DHM in the liver alcohol elimination at its molecular level. First, impact of DHM on alcohol dehydrogenase (ADH) activity in vitro and the enzyme induction in vivo was examined. Neither the ADH activity nor the enzyme expression were influenced by DHM. Next, the effect of DHM during alcohol intoxication were studied on primary hepatocytes isolated from EtOH-premedicated and untreated rats. The viability of cells exposed to alcohol, estimated based on the released enzymes, alanine aminotransferase (ALT) and aspartate aminotransferase (AST), was slightly affected by DHM. Although the expected hepatoprotective effect of DHM was not fully achieved, DHM (in a concentration manner) proved to reduce the level of ROS/RNS in hepatocytes. However, no change in the rate of alcohol metabolism in vivo was found when rats were administered with a single or repeated dose of ethanol supplemented with DHM. In conclusion, the proposed positive effect of DHM during alcohol intoxication has not been proven. Moreover, there is no effect of DHM on the alcohol metabolism. The "hoped-for" DHM hepatoprotective activity can be attributed to the reduction of ROS/RNS levels in cells.

Department of Biochemistry Faculty of Science Charles University Prague 2 Czech Republic

Zobrazit více v PubMed

CHEN X, ZHONG Z, XU Z, CHEN L, WANG Y. 2′,7′-Dichlorodihydrofluorescein as a fluorescent probe for reactive oxygen species measurement: Forty years of application and controversy. Free Radic Res. 2010;44:587–604. doi: 10.3109/10715761003709802. PubMed DOI

FARGHALI H, BUCHAR E, MACHKOVA Z, KAMENIKOVA L, MASEK K. Muramyl dipeptide and carbon tetrachloride hepatotoxicity in rats: involvement of plasma membrane and calcium homeostasis in protective effect. Methods Find Exp Clin Pharmacol. 1986;8:469–477. PubMed

HARTREE EF. Determination of protein: a modification of the Lowry method that gives a linear photometric response. Anal Biochem. 1972;48:422–427. doi: 10.1016/0003-2697(72)90094-2. PubMed DOI

HODEK P, KRIZKOVA J, FREI E, SINGGH R, ARTL VM, STIBOROVA M. Impact of beta-naphthoflavone on genotoxicity of food-derived carcinogens. Neuro Edocrinol Lett. 2011;32(Suppl 1):25–34. PubMed

HOU X, TONG Q, WANG W, XIONG W, SHI C, FANG J. Dihydromyricetin protects endothelial cells from hydrogen peroxide-induced oxidative stress damage by regulating mitochondrial pathways. Life Sci. 2015;130:8–46. doi: 10.1016/j.lfs.2015.03.007. PubMed DOI

JIANG L, ZHANG Q, REN H, MA S, LU CJ, LIU B, LIU J, LIANG J, LI M, ZHU R. Dihydromyricetin enhances the chemo-sensitivity of nedaplatin via regulation of the p53/Bcl-2 pathway in hepatocellular carcinoma cells. PloS One. 2015;10:e0124994. doi: 10.1371/journal.pone.0124994. PubMed DOI PMC

KRIZKOVA J, BURDOVA K, HUDECEK J, STIBOROVA M, HODEK P. Induction of cytochromes P450 in small intestine by chemopreventive compounds. Neuro Endocrinol Lett. 2008;29:717–721. PubMed

LeBEL CP, ISCHIROPOULOS H, BONDY SC. Evaluation of the probe 2',7'-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem Res Toxicol. 1992;5:227–231. doi: 10.1021/tx00026a012. PubMed DOI

LIANG J, OLSEN RW. Alcohol use disorders and current pharmacological therapies: the role of GABAA receptors. Acta Pharmacol Sin. 2014;35:981–993. doi: 10.1038/aps.2014.50. PubMed DOI PMC

LIU B, TAN X, LIANG J, WU S, LIU J, ZHANG Q, ZHU R. A reduction in reactive oxygen species contributes to dihydromyricetin-induced apoptosis in human hepatocellular carcinoma cells. Sci Rep. 2014;4:7041. doi: 10.1038/srep07041. PubMed DOI PMC

LIU Z, YANG K, CHEN Z, CHENG Q, WU L. The versatile effects of dihydromyricetin in health. Evid Based Complement Alternat Med. 2017;6:1053617. doi: 10.1155/2017/1053617. PubMed DOI PMC

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–275. PubMed

MOLDEUS P, HOGBERG J, ORRENIUS S. Isolation and use of liver cell. Methods Enzymol. 1978;52:60–71. doi: 10.1016/S0076-6879(78)52006-5. PubMed DOI

MOSMANN T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55–63. doi: 10.1016/0022-1759(83)90303-4. PubMed DOI

MURAKAMI T, MIYAKOSHI M, ARAHO D, MIZUTANI K, KAMBARA T, IKEDA T, CHOU WH, INUKAI M, TAKENAKA A, IGARASHI K. Hepatoprotective activity of tocha, the stems and leaves of Ampelopsis grossedentata, and ampelopsin. Biofactors. 2004;21:175–178. doi: 10.1002/biof.552210136. PubMed DOI

PETER R, BÖCKER R, BEAUNE PH, IWASAKI M, GUENGERICH FP, YANG CS. Hydroxylation of chlorzoxazone as a specific probe for human liver cytochrome P-450IIE1. Chem Res Toxicol. 1990;3:566–573. doi: 10.1021/tx00018a012. PubMed DOI

SHEN Y, LINDEMEYER AK, GONZALEZ C, SHAO XM, SPIGELMAN I, OLSEN RW, LIANG J. Dihydromyricetin as a novel anti-alcohol intoxication medication. J Neurosci. 2012;32:90–401. doi: 10.1523/JNEUROSCI.4639-11.2012. PubMed DOI PMC

SILVA J, YU X, MORADIAN R, FOLK C, SPATZ MH, KIM P, BHATTI AA, DAVIES DL, LIANG J. Dihydromyricetin protects the liver via changes in lipid metabolism and enhanced ethanol metabolism. Alcohol Clin Exp Res. 2020;44:1046–1060. doi: 10.1111/acer.14326. PubMed DOI PMC

STROBER W. Trypan blue exclusion test of cell viability. Curr Protoc Immunol. 2001;111:3. doi: 10.1002/0471142735.ima03bs21. PubMed DOI

ZAKHARI S. Overview: how is alcohol metabolized by the body? Natl Inst Alcohol Abuse Alcohol. 2006;29:245–254. doi: 10.1159/000095013. PubMed DOI PMC