title: Hugan Qingzhi medication ameliorates hepatic steatosis by activating AMPK and PPARα pathways in L02 cells and HepG2 cells.
abstract: BACKGROUND
  Hugan Qingzhi tablet (HQT), a lipid- lowering traditional Chinese medicine formula, has been used for the prevention and treatment of nonalcoholic fatty liver (NAFLD).
  
  
  OBJECTIVE
  This study was realized to evaluate the effects of HQT-medicated serum on hepatic steatosis using in vitro experiments with cells and explore the relevant mechanisms with method of serum pharmacology.
  
  
  METHODS
  A model of hepatic steatosis in the L02 and HepG2 cells was induced by free fatty acid (FFA). The components in the HQT-medicated serum were assayed by high-performance liquid chromatography. Intracellular lipid droplets were detected by Oil Red O staining, and their ultrastructure was examined by transmission electron microscope. The biochemical parameters, including triglyceride (TG), lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT), total antioxidant capacity (T-AOC), malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH), were measured with commercial kits. Furthermore, the expression of adiponectin, AMP-activated protein kinase (AMPK) phosphorylation, sterol regulatory element-binding protein 1 (SREBP-1), peroxisome proliferator activated receptor-α (PPARα), carnitine palmitoyltransferase 1 (CPT-1), and acetyl-CoA oxidase 1 (ACOX1) was analyzed by Western blot and/or quantitative reverse transcription-polymerase chain reaction (qRT-PCR).
  
  
  RESULTS
  Moderate- and high-dose HQT-medicated serum reduced (P<0.05 or P<0.01) the accumulation of lipid droplets and the cellular TG content in L02 and HepG2 cells. They caused significant reductions (P<0.01) in LDH, AST, ALT and MDA and significant increase (P<0.05 or P<0.01) in T-AOC in the culture medium. They also caused increase (P<0.05 or P<0.01) in GSH level and SOD activity in FFA-induced steatotic L02 and HepG2 cells. Furthermore, moderate- and high-dose HQT-medicated serum enhanced (P<0.01) adiponectin expression in a concentration-dependent manner and increased (P<0.05 or P<0.01) the phosphorylation of AMPK and the expression of PPARα, CPT-1, and ACOX1, and reduced (P<0.05 or P<0.01) the expression of SREBP-1.
  
  
  CONCLUSIONS
  The results suggested that HQT-medicated serum exerts a preventive effect against hepatic steatosis, and the potential mechanism might be activation of AMPK and PPARα pathways.
authors.affiliation: Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou 510282, PR China.
authors.forename: JinJin
authors.initials: J
authors.lastname: Yin
authors.affiliation: Department of Pharmacy, Nanfang Hospital, Southern Medical University, Guangzhou 510515, PR China.
authors.forename: YanQin
authors.initials: Y
authors.lastname: Luo
authors.affiliation: Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou 510282, PR China.
authors.forename: HouLiang
authors.initials: H
authors.lastname: Deng
authors.affiliation: Institute of Digestive Disease, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, PR China.
authors.forename: ShuMin
authors.initials: S
authors.lastname: Qin
authors.affiliation: Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou 510282, PR China.
authors.forename: WaiJiao
authors.initials: W
authors.lastname: Tang
authors.affiliation: Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou 510282, PR China.
authors.forename: Lu
authors.initials: L
authors.lastname: Zeng
authors.affiliation: Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Guangdong, Guangzhou 510282, PR China. Electronic address: zhoubj163@163.com.
authors.forename: BenJie
authors.initials: B
authors.lastname: Zhou
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000378:metabolism*; D052242:Adiponectin / Q000378:metabolism*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology* / Q000627:therapeutic use; D002460:Cell Line; D004365:Drugs, Chinese Herbal / Q000494:pharmacology* / Q000627:therapeutic use; D005234:Fatty Liver / Q000188:drug therapy / Q000378:metabolism*; D005260:Female; D056945:Hep G2 Cells; D006801:Humans; D019301:Oleic Acid; D018384:Oxidative Stress / Q000187:drug effects; D047493:PPAR alpha / Q000378:metabolism*; D019308:Palmitic Acid; D017207:Rats, Sprague-Dawley
chemical_list: C499918:ADIPOQ protein, human; D052242:Adiponectin; D000975:Antioxidants; D004365:Drugs, Chinese Herbal; D047493:PPAR alpha; C000594496:hugan qingzhi; D019301:Oleic Acid; D019308:Palmitic Acid; D055372:AMP-Activated Protein Kinases
keywords: 23-O-acetylalisol B (PubChem CID: 3084460); AMPK; Adiponectin; Hepatic steatosis; Hugan Qingzhi tablet; Isorhamnetin-3-O-neohesperidoside (PubChem CID: 24204448); Nuciferine (PubChem CID: 10146); Oxidative stress; PPARα; Quercetin (PubChem CID: 5280343); Ursolic acid (PubChem CID: 64945)