| Objective To explore the effect of leptin to triglyceride's disposition and expression of acetyl-CoA carboxylasemRNA in normal and nonalcoholic induced fatty liver cell.Methods The investigation was conducted by means of randomized controlled trial. To establish a model of hepatocyte steatosis, we incubate the normal human L-02 hepatocyte in RPMI-1640 complete medium with 50% fetal bovine serum for 24 hours. Then add recombinant human leptin of varies concentration into the medium and incubate for another 24 hours. The concentration of leptin was respectively 10-6mol/l, 10-6mol/l and 10-7mol/l. And observe the formation of cellular morphology and lipid droplets in the cells under oil red 0 stain and determin the intracellular triglyceride levels through high performance liquid chromatography (HPLC). The RT-PCR was used to detect the mRNA levels of acetyl-CoA carboxylase of each group. The best concentration of leptin that decreases the amount of intracellular triglyceride levels was decided and the dose-effect relationship curve was established. Then expose the cell to leptin at the optimal concentration for 6h, 12h, 24h respectively, observe the above-mentioned parameters and determine the best duration for leptin to decrease intracellular triglyceride levels, and draw out the duration-effect relation curve.Results Oil red staining show that 50% inactivation embryo blood serum can make fatty liver cell model for 24 hours .After L-02 liver cell were turned into fatty liver cell , RT- PCR show that acetyl-CoA carboxylasemRNA expression increased obviously. When give leptin to fatty liver cell, The HPLC show that model group triglyceride's disposition increased clearly, but were given leptin to model cells, triglyceride's disposition decreased obviously and the variation was positive... |
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