Adipocyte Differentiation Related Gene Expression Of LXRαand Its Primary Analysis By Bioinformatics

Adipocytes differentiation is a expression process of somemarker genes really. The series of genes include PPARγ, HSL andLPL. Liver X receptorα(LXRα) gene is a member of the nuclearreceptor supper family, which play a dominant role in lipidmetabolism. LXRαis highly expressed in adipocyte. And it canenhance the expression of adipocyte specific genes. For examplePPARr, LPL and leptin. Therefore, we infer LXRαmay haveparticipated during adipocytes differentiation. Eytokines areimportant factors affecting adipocytes differentiation. Someresearches indicated IL-6 can stimulate lipolysis and fatty acidoxidation. While at present，the molecular mechanism is remainsunclear for IL-6 regulating of adipocyte differentiation.In this study，at first, we analyzed the the pig LXRαbybioinformatics methods which has been published on gene bank. Then healthy pigs were used as experimental animal. The mRNAdistribution profile of the porcine LXRαgene in eight tissues (heart,liver, subcutaneous adipose, kidney, visceral adipose, spleen, lung,Muscle) from the pig sample were examined by RT-PCR. Then the SDrats were incubated with different IL-6 concentrations for varioustimes Afterthat, RNA and protein were extracted from adiposetissue. Further, gene expression of PPARr, LXRα, LPL snd HSL weredetermined with RT-PCR and Western blot to resolve the possiblemechanisms of IL-6 regulating of adipocyte differentiation. It willprovide available information for control of human obesity and thetherapy of metabolic diseases.The main results were as follows:1. Protein structure analysis suggested that that the relativemolecular weight of LXRαreceptor is 502910.00 D, coding 454Amino Acids, LXRαprotein is mainly consists of three functionaldomains: a Ligand-binding Domain, a DNA-binding Domain, and aZinc fingerr C4 region. phylogenetic result showed that the LXRαgenes in Mammalia clade were very conserved.2. When the annealing temperature is 48.9℃and MgCl2concentration is 2mmol/L, the pig LXRαgene was amplificatedefficiently. 3. The mRNA distribution profile of the porcine LXRαgene in eighttissues (heart, liver, subcutaneous adipose, kidney, visceral adipose,spleen, lung, Muscle) from the pig sample were examined by RT-PCR.The results showed that the LXRαgene was expressed specificity.The porcine LXRαgene highly expressed in liver, visceral adiposetissue, subcutaneous adipose tissue and kidney. While remarkablelower in heart, lung, spleen and muscle. The highest of LXRαexpression levels were found in liver tissue. The results suggestedthat LXRαhas a close correlation with lipid metabolism.4. Then the SD rats were incubated with different IL-6concentrations (0, 0.1ug/ml, 0.4ug/ml) for 48h. The result of RT-PCRshow that IL-6 significantly inhibits the mRNA expression ofadipogenic marker genes PPARr, LXRαand LPL. But HSL mRNAlevels were not changed. The effect of IL-6 on expresion of PPARγwas detected by Western blot. All these test support the downregulation of protein expression for PPARγby IL-6.These results showed that different concentrations of IL-6 couldinhibit adipocyte differentiation by regulating the expressions ofLXRα, LPL and PPARγ.