The Influence Of Wnt/β-catenin Signaling Agonist-LiCl For Chemerin Gene Expression In 3T3-L1 Preadipocyte Differentiation

Obesity, now as the factors that increasingly affects human health, has become one of thediseases affect the quality of human life worldwide. Obese people, mostly accompanied by avariety of complications, such as diabetes, atherosclerosis, cardiovascular diseases, andattention has been paid. Therefore, to study of obesity in the pathogenesis, and find the entrypoint for the treatment of obesity become problems which to be solved in the field of scientificresearch and even the medical field today.Differentiation of preadipocyte plays a facilitating roll for fat accumulation undoubtedly.Fat cell differentiation, but also gradually becomes one of the hot research obesity. In vitro,the 3T3-L1 preadipocytes are induced with drugs incubation into adipocytes, which mimickedadipocyte differentiation process in vivo, and this model provides a good platform for manyobesity researchers.Now, the role of many key genes and important factors for the 3T3-L1 differentiationmechanism has gradually become clear. For example, both the peroxisome proliferator-activatedreceptorγand the CCAAT / enhancer binding protein have been considered the key factor forcontrol of adipocyte differentiation. In recent years, inhibition of Wnt /beta-catenin signalingpathway in adipocyte differentiation has also been confirmed by numerous institute. As a newlydiscovered adipokine, chemerin expression changes in adipocyte differentiation has becomea recent focus of attention. For adipocyte differentiation, the growing body of research has grownfrom a single gene, multiple genes or even multi-factor interactions tend to the exploration of thedialogue between the signaling pathways, however, between the Wnt / beta-catenin signalingpathway and chemerin, whether there is a certain relationship is not yet any reports.Our study was on these two aspects: the expression of the Wnt /beta-catenin signalingpathway excited by LiCl in the process of cell differentiation of 3T3-L1 adipocytes, usingRT-PCR technique to detect chemerin expression changes, as well as LiCl concentration gradientintervention experiment. The results showed that: 1) By using hormone "cocktail" induced 3T3-L1 preadipocytes and after that stained cells by oilred O. The induction rate reached 85%~90%, successfully established a fat differentiationmodel, and laid the foundation for subsequent experiments.2) In the experimen of exploring LiCl intervention concentration, it found that 10 mM wasnot able to completely inhibit the differentiation of cells; 20mM totally inhibited thedifferentiation and fully activated the expression of Wnt / beta-catenin signaling pathway,so the follow-up experiment using 20mM of LiCl.3)Using 20mM of LiCl in the process of differentiation of 3T3-L1cells, oil red O resultsshowed that: The coloring of non-intervention group of lipid droplets at Day3 beganto appear more and more with the differentiation process, maximum to Day14. Theintervention group coloring lipid droplets in Day14 just a little.4) Using 20mM of LiCl incubated in the process of differentiation of 3T3-L1 cells, RT-PCRresults showed that: the chemerin of RNA level of non-intervention group from Day0 toDay1 was down, and thereafter gradually increased with the differentiation process, whilethe intervention group chemerin also showed similar tends, but its expression was inhibitedto a large extent.5) In 3T3-L1 cells differentiation and maturation, the LiCl concentration gradient interventedfor 24hours, which were setup 0,100 uM, 1mM, 5mM, 10mM, 20mM six concentration. Theresults showed that: even in the mature fat cells, RNA expression of LiCl on chemerin stillwas inhibited. And the inhibitory effect was becoming more apparent with increasing LiClconcentration. But the effect was not as good as the pre-intervention.The above results showed that the intervention of LiCl inhibited the expression of chemerin. Thisinhibition performance in two ways: First, adding in the differentiation of 3T3-L1 cells process of LiCl,showed the significant inhibitory effect on chemerin; LiCl incubated for 24 hours in 3T3-L1 cellsdifferentiation and maturation, inhibitory effect on chemerin was present but not obvious. The resultssuggested that Wnt/beta-catenin signaling pathway may have an effect on the expression ofchemerin which was direct or indirect role still had yet to be further research.