Real-time quantitative PCR analysis of mouse liver tissue and liver cells

Diets rich in fat is a major cause of common chronic liver diseases in worldwide and nutritional food has been widely used to counteract the metabolic disorders such as non-alcoholic fatty liver disease (NAFLD). The present study investigated the effects of oleuropein-enriched extract from Jasminum grandiflorum L. flowers (OLE-JGF), a medicinal plant that is also widely consumed as a beverage in China, in high fat diet (HFD) fed mice and oleic acid (OA)-treated AML-12 cells. Treatment of HFD-fed mice with 0.6% (w/w) OLE-JGF for 8 weeks significantly reduced the plasma levels of ALT, TC and LDL-C without changing the liver weight. Additionally, OLE-JGF administration significantly suppressed the mRNA expressions of the inflammatory mediators MCP-1 and CD68 in the liver of HFD-fed mice. Importantly, treatment with OLE-JGF significantly downregulated the key lipogenic enzymes (ACC and FAS) and their upstream transcription factor SREBP-1c in the liver. In the same time, mitochondrial DNA and UCP2 were upregulated along with increased expression of mitochondrial biogenic promoters including LKB1 and its downstream gene PGC-1α, Nrf2 and Tfam, but not changed AMPK in liver. In vitro, treatment of OLE for 24 h significantly increased the cell viability and decreased the TG level in OA-induced AML-12 cells. OLE significantly inhibited ACC mRNA expression and upregulated LKB1, PGC-1α and Tfam mRNA levels in OA-treated cells. In addition, OLE significantly increased the binding level of LKB1 to PGC-1α promoter in OA-induced cells. These findings indicate that OLE-JGF attenuates hepatocyte damage in HFD-fed mice and OA-induced liver cells, may be partly attributed to upregulation of LKB1-PGC-1α axis, which mediates hepatic lipogenesis and mitochondrial biogenesis. Our study provides a scientific basis for the benefits of J. grandiflorum flower as a supplement for met-abolic disease and potential use of OLE for the treatment of chronic liver diseases. qPCR gene expression profiling. Equal amount total RNA from each donor was pooled prior to gene expression analysis.Animal experiment Chow, HFD and HFD-JGF groups of 4-9 mice in each group were used as independent replicates. Cell test Control, OA, OLE three groups each more than or equal to three independent repeated test