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Kaempferol-3-O-Glucuronide Ameliorates Non-Alcoholic Steatohepatitis in High-Cholesterol-Diet-Induced Larval Zebrafish and HepG2 Cell Models via Regulating Oxidation Stress

Authors

Keywords:

non-alcoholic fatty liver disease, kaempferol-3-O-glucuronide, oxidative stress, inflammation, high-cholesterol diet.

Abstract

NAFLD (non-alcoholic fatty liver disease) is one of the most prominent liver diseases in the world. As a metabolic-related disease, the development of NAFLD is closely associated with various degrees of lipid accumulation, oxidation, inflammation, and fibrosis. Ilex chinensis Sims is a form of traditional Chinese medicine which is used to treat bronchitis, burns, pneumonia, ulceration, and chilblains. Kaempferol-3-O-glucuronide (K3O) is a natural chemical present in Ilex chinensis Sims. This study was designed to investigate the antioxidative, fat metabolism-regulating, and anti-inflammatory potential of K3O. A high-cholesterol diet (HCD) was used to establish steatosis in larval zebrafish, whereby 1mM free fatty acid (FFA) was used to induce lipid accumulation in HepG2 cells, while H2O2 was used to induce oxidative stress in HepG2. The results of this experiment showed that K3O reduced lipid accumulation and the level of reactive oxygen species (ROS) both in vivo (K3O, 40 μM) and in vitro (K3O, 20 μM). Additionally, K3O (40 μM) reduced neutrophil aggregation in vivo. K3O (20 μM) also decreased the level of malondialdehyde (MDA) and significantly increased the level of glutathione peroxidase (GSH-px) in both the HCD-induced larval zebrafish model and H2O2-exposed HepG2 cells. In the mechanism study, keap1, nrf2, tnf-α, and il-6 mRNA were all significantly reversed by K3O (20 μM) in zebrafish. Changes in Keap1 and Nrf2 mRNA expression were also detected in H2O2-exposed HepG2 cells after they were treated with K3O (20 μM). In conclusion, K3O exhibited a reduction in oxidative stress and lipid peroxidation, and this may be related to the Nrf2/Keap1 pathway in the NAFLD larval zebrafish model. Keywords: non-alcoholic fatty liver disease, kaempferol-3-O-glucuronide, oxidative stress, inflammation, high-cholesterol diet.

Author Biographies

Yang Deng, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Ji Ma, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Xin Weng, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Yuqin Wang, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Maoru Li, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Tingting Yang, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Zhiyang Dou, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Zhiqi Yin, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

Jing Shang, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198, China; [email protected] (Y.D.); [email protected] (J.M.); [email protected] (X.W.); [email protected] (Y.W.); [email protected] (M.L.); [email protected] (T.Y.); [email protected] (Z.D.)

State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China

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