文献类型： 外文期刊

作者： Sun, Binmei ¹ ; Tan, Deguan ¹ ; Pan, Dongjin ¹ ; Baker, Margaret R. ¹ ; Liang, Zhibin ¹ ; Wang, Zhizheng ⁶ ; Lei, Jianjun ² ; Liu, Shaoqun ² ; Hu, Ching Yuan ⁷ ; Li, Qing X. ¹ ;

作者机构： 1.Univ Hawaii Manoa, Dept Mol Biosci & Bioengn, Honolulu, HI 96822 USA

2.South China Agr Univ, Coll Hort, Guangzhou, Peoples R China

3.Chinese Acad Trop Agr Sci, Minist Agr, Inst Trop Biosci & Biotechnol, Key Lab Trop Crops Biol & Genet Resources, Haikou, Hainan, Peoples R China

4.Chinese Acad Trop Agr Sci, Hainan Acad Trop Agr Resource, Haikou, Hainan, Peoples R China

5.Guangxi Univ Chinese Med, Inst Marine Drugs, Nanning, Guangxi, Peoples R China

6.Cent China Normal Univ, Coll Chem, Key Lab Pesticide & Chem Biol, Minist Educ, Wuhan, Peoples R China

7.Univ Hawaii Manoa, Dept Human Nutr Food & Anim Sci, Honolulu, HI 96822 USA

8.Salk Inst Biol Studies, Cellular Neurobiol Lab, 10010 N Torrey Pines Rd, La Jolla, CA 92037 USA

9.Salk Inst Biol Studies, Paul F Glenn Ctr Biol Aging Res, 10010 N Torrey Pines Rd, La Jolla, CA 92037 USA

关键词： adipocyte; antiadipogenesis; drug affinity responsive target stability (DARTS); 78-kDa glucose-regulated protein; GRP78; lipid droplet; obesity; proteomics; surface plasmon resonance

期刊名称：JOURNAL OF NUTRITION （ 影响因子：4.687； 五年影响因子：5.76 ）

ISSN： 0022-3166

年卷期： 2021 年 151 卷 7 期

页码：

收录情况： SCI

摘要： Background: Obesity is among the most serious public health problems worldwide, with few safe pharmaceutical interventions. Natural products have become an important source of potential anti-obesity therapeutics. Dihydromyricetin (DHM) exerts antidiabetic effects. The biochemical target of DHM, however, has been unknown. It is crucial to identify the biochemical target of DHM for elucidating its physiological function and therapeutic value. Objectives: The objective of this study was to identify the biochemical target of DHM. Methods: An abundant antiadipogenic flavanonol was extracted from the herbal plant Ampelopsis grossedentata through bioassay-guided fractionation and characterized with high-resolution LC-MS and H-1 and C-13 nuclear magnetic resonance. Antiadipogenic experiments were done with mouse 3T3-L1 preadipocytes. A biochemical target of the chemical of interest was identified with drug affinity responsive target stability assay. Direct interactions between the chemical of interest and the protein target in vitro were predicted with molecular docking and subsequently confirmed with surface plasmon resonance. Expression levels of peroxisome proliferator-activated receptor gamma (PPAR gamma), which is associated with 78-kDa glucose-regulated protein (GRP78), were measured with real-time qPCR. Results: DHM was isolated, purified, and structurally characterized. Cellular studies showed that DHM notably reduced intracellular oil droplet formation in 3T3-L1 cells with a median effective concentration of 294 mu M (i.e., 94 mu g/mL). DHM targeted the ATP binding site of GRP78, which is associated with adipogenesis. An equilibrium dissociation constant between DHM and GRP78 was 21.8 mu M. In 3T3-L1 cells upon treatment with DHM at 50 mu M (i.e., 16 mu g/mL), the expression level of PPAR gamma was downregulated to 53.9% of the solvent vehicle control's level. Conclusions: DHM targets GRP78 in vitro. DHM is able to reduce lipid droplet formation in 3T3-L1 cells through a mode of action that is plausibly associated with direct interactions between GRP78 and DHM, which is a step forward in determining potential applications of DHM as an anti-obesity agent.