Hugan Qingzhi medication ameliorates hepatic steatosis by activating AMPK and PPAR伪 pathways in L02 cells and HepG2 cells

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• 作者：JinJin Yin^a ; YanQin Luo^b ; HouLiang Deng^a ; ShuMin Qin^c ; WaiJiao Tang^a ; Lu Zeng^a ; BenJie Zhou^a ; ^{zhoubj163@163.com" class="auth_mail} ; ^{yinjinjinsunny@163.com" class="auth_mail}
• 关键词：ACOX1 ; acetyl-CoA oxidase 1 ; ALT ; alanine aminotransferase ; AMPK ; AMP-activated protein kinase ; AST ; aspartate aminotransferase ; ChREBP ; carbohydrate-responsive element-binding protein ; CPT-1 ; carnitine palmitoyltransferase 1 ; FFA ; free fatty acid ; GSH ; glutathione ; HPLC ; high-performance liquid chromatography ; HQT ; Hugan Qingzhi tablet ; LDH ; lactate dehydrogenase ; MDA ; malondialdehyde ; NAFLD ; nonalcoholic fatty liver disease ; NASH ; non-alcoholic steatohepatitis ; PPAR伪 ; peroxisome proliferator a
• 刊名：Journal of Ethnopharmacology
• 出版年：28 May 2014
• 年：2014
• 卷：154
• 期：1
• 页码：229-239
• 全文大小：4332 K

文摘

Hugan Qingzhi tablet (HQT), a lipid- lowering traditional Chinese medicine formula, has been used for the prevention and treatment of nonalcoholic fatty liver (NAFLD).

Aim of the study

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.

Materials and 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-伪 (PPAR伪), 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 PPAR伪, CPT-1, and ACOX1, and reduced (P<0.05 or P<0.01) the expression of SREBP-1.

Conclusion

The results suggested that HQT-medicated serum exerts a preventive effect against hepatic steatosis, and the potential mechanism might be activation of AMPK and PPAR伪 pathways.