Effect Of Testosterones On MRNA Expression Of Leptin And Adiponectin Shown By Human Omental Preadipocytes

Objective:To investigate the effect of testosterone on leptin and adiponectin mRNA expressions in adipocytes in human omental preadipocytes.Methods:Omental adipose tissue biopsies about 5-10g were obtained from patients undergoing elective open-abdominal surgery.Preadipocytes were incubated and then differentiated in vitro into mature adipocytes.Morphology and extracting stained intracytoplasmic lipid with oil red O all verified their preadipocyte identity.These cells were incubated for 24 hours in the presence of 10-9mol/L,10-8mol/L or 10-7mol/L testosterone respectively.Adipocytes were stimulated by 10-7mol/L testosterone for 12 hours,24 hours or 48 hours respectively.Total RNA isolated was evaluated, and then, the qualified RNA was carried out by reverse transcription- polymerase chain reaction(RT-PCR).GAPDH was used to be inside reference. Then leptin mRNA, adiponectin mRNA and GAPDH mRNA were amplified by specifical primer. After 3.0% agarose gel electrophoresis, the ratio of leptin mRNA /GAPDH mRNA and adiponectin mRNA /GAPDH mRNA were detected by gel image analysis technology and analyzed. Statistical analysis was undertaken using a one-way ANOVA for comparison of control versus treatments groops. Data in the text and figures are expressed as means±SE.The threshold for significance was P<0.05.Results:1.Human preadipocytes were primary cultured successfully and then differentiated in vitro into mature adipocytes.2.Leptin mRNA and adiponectin mRNA were decreased when the cells incubated with testosterone (10-9mol/L-10-7mol/L,respectively) and leptin mRNA and adiponectin mRNA were decreased when the cells incubated with 10-7mol/L testosterone (12h-48h,respectively).Conclusion:Human preadipocytes were primary cultured successfully.This study lay the basis for further probing into the mechanisms which all kinds of hormones effect adipokines.testosterone can decreased leptin mRNA and adiponectin mRNA expression in human omental adipocytes suggesting that were evidence about the sex specificities in 2 diebetes and obesity.