The Research Of The Lipogenesis Of Hepatocyte Induced By SREBP-2

Liver is an important organs that gives priority to the metabolism in human body,and it has many important functions including removing the toxins, glycogen storage,secreted protein synthesis and so on. When lipid metabolism disorders happen in the cell, many diseases occur, such as non-alcoholic fatty liver disease and so on. Liver lipid metabolism is a complicated dynamic balance process regulating by many factors. At the present, the SREBPs family are recognized key transcription factors that regulate lipid metabolism, and they play importantly regulative roles during the lipid metabolism process. However, the specific regulation mechanism is still unclear.To further search for the process and molecular mechanism of liver lipid metabolism regulated by SREBP-2, we firstly used the adult mice as the liver cell donors and separated the primary hepatocytes using enzyme digestion method, and then preliminary identified the primary cells through observation and PAS methods nfirming that the isolated primary cells were hepatocytes. Lastly, we used the Recombinant adenovirus carrying SREBP-2 to infect hapetacytes, and detected the relative transcript level of SREBP-2,C/EBPα,FAS and SREBP-1c using real-time PCR to analyze the molecular mechanism of liver lipid metabolism regulated by SREBP-2。The main results were as follows:(1)We successfully isolated the high survival rate primary hepatocytes. The cells were dyed red using PAS method,and cell morphology was uniform, indicating that cells contained a lot of liver glycogen and the purity of isolated hepatocytes was higher.(2) Real-time PCR results indicated Ad-SREBP-2 adenovirus can enhanced the mRNA expression of SREBP-2 in the primary hepatocytes by increasing the expression more than 80 times.(3) Oil O staining results demonstrated that over-expression of SREBP-2 could promote the synthesis and deposition of lipid droplets of hepatocytes.(4) Over-expression of SREBP-2 significantly promoted the expression of SREBP-1,while it had no significant effect on the expression of C/EBPα.In conclusion, we successfully isolated and identified the hepatocytes, and found that over-expression of SREBP-2 could significantly promoted deposition of lipid droplets and the high expression of lipid synthesis related genes, suggesting that SREBP-2 has an important regulatory effects on the lipid synthesis process of hepatocytes, and might be a key regulatory factor of hepatic lipid metabolism disorders.