Real-time quantitative PCR analysis of mouse liver tissue and liver cells. 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. Overall design: 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