Chemerin Expression Changes In 3t3-l1 Adipocyte Differentiation Process And The Relationship Between Adiponectin, Leptin Preliminary Study

Objective:1.To observe the change of chemerin and chemerinR gene expression in various tissues of mice and in the process of 3T3-L1 preadipocyte differentiation.2.To explore the effects of PPAR-y agonist-rosiglitazone on chemerin and chemerinR gene expression.3.To observe the effects of recombinant mouse chemerin on 3T3-L1 preadipocyte differentiation and the change of adiponectin.leptin and PPAR-γ2 gene expression.Method:1. Total RNA was extracted from several kinds of tissues (liver, adipose tissue, stomach, spleen, kidney, heart and skeletal muscle) and Semi-quantitative RT-PCR was performed in order to measure levels of chemerin and chemerinR mRNA.Culture of 3T3-L1 cells and induction of differentiation were performed in vitro. Realtime-PCR was performed in order to measure levels of chemerin and chemerinR mRNA of differential time points (0d,2d,4d,6d,8d,10d).2.3T3-L1 cells were differentiated for 10 days as the normal control group; the other group in the induction of differentiation using 10μM rosiglitazone. Real-time fluorescent quantitative PCR method was applyed to detect the expressions of chemerin and chemerinR gene. After 3T3-L1 fat cells were matured, different concentrations of rosiglitazone(0,0.1,1,10μM) interfered for 24 hours then collected the cells, real-time quantitative PCR was used to detect the expression of chemerin and chemerinR gene.3T3-L1 cells differentiated for 10 days as the normal control group; a process of induced differentiation using rosiglitazone; another group using chemerin protein intervention. Real-time quantitative PCR was used to detect the expressions of PPAR-γ2 which was in the tenth days, so were C/EBPa gene and adiponectin, leptin gene. After 3T3-L1 fat cells were matured, different concentrations of chemerin protein interfered for 24 hours then collected the cells, real-time quantitative PCR was used to detect the expression of PPAR-γ2,C/ EBPa,adiponectin, leptin gene.Results:1. chemerin and chemerinR gene were expressed in liver, adipose tissue, stomach, spleen, kidney, heart and skeletal muscle of mice. Chemerin and chemerinR mRNA levels dramatically increased during the process of differentiation of 3T3-L1 cells after 2 days of induction. (P<0.05).2. Compared with the control group, during 3T3-L1 preadipocytes differentiation and matured 3T3-L1 adipocytes,chemerin and chemerinR gene were upregulated using rosiglitazone (P<0.05).3.In 3T3-L1 cells during differentiation for 10 days compared with the control using rosiglitazone group, chemerin protein group could raised PPAR-γ2, C/EBPa, adiponectin, leptin gene expression (p<0.05). In matured 3T3-L1 cells, compared with the control group, chemerin protein raised PPAR-γ2, C/EBPa and adiponectin, leptin gene expression (p <0.05).Conclusion:1. chemerin and chemerinR were both expressed in adipose tissues of mice; the expressions of chemerin and chemerinR gene were gradually increasesd during the 3T3-L1 preadipocyte differentiation process.2. PPAR-y agonist Rosiglitazone upregulated the expressions of chemerin and chemerinR gene.3. chemerin protein upregulated the expressions of PPAR-γ2, C/EBPa and adiponectin, leptin gene in matured 3T3-L1 adipocytes.