The Preliminary Study Of The Effect Of Piogliazation On 3T3 L1 Preadipocyte

Objective: To observe the effect of pioglitazone on preadipocyte proliferation and differentiation, detect apelin concentration of the cell supernatant after intervention, and to investigate the mechanisms of thiazolidinedione affect endocrine function of adipose tissue.Method: Preadipocytes were serperately cultured by the basic medium with four different consistences of pioglitazone in vitro, then detected apelin concentration of the cell supernatant by ELISA after 48 hours, and evaluated cell proliferation by MTT test, to find an effective concentration of pioglitazone. According the different medium, the preadipocyte devided into three groups including control group A, differentiation group B, experimental group C. At the varied experimental phases of 24, 48, 72 hours, the apelin concentration of supernatant was detected, the cell parameters of proliferation and differentiation were observed. In 5 and 10 days, the absorbance of cells stained with oil red O was detected. The apelin concentrations,and parameters of preadipocyte proliferation and differentiation of the three groups were evaluation.Results: 1. Apelin could be detected in the supernatant of A, B and C group, and its concentration was changed with the culture time,at 72 hours the apelin levels in B, C group remarkly increased by 33.7%, 32.5% than A group (P<0.05). 2. The effect on cell differentiation: oil red O staining test showed that the absorbance values of the 10μmol/L, 100μmol/L pioglitazone groups were greater than the control group. At 24,48 hours, the absorbance of C group staining with oil red O had no significant difference compared with B group; At 72 hours, the absorbance value of C group was higher than the value of A group, the difference was statistically significant (P<0.05), but lower than B group; in the first 10 days,the difference between B and C group was not significant (P≥0.05), but the difference was still significant between A and C group(P<0.05). After 3 days, some small lipid droplets were founded in cytoplasmic of preadipocyte cultrued in C group, in 5, 10 days when the promotion of the cytoplasm of fat cells is polyester, the results were compared with A group increased by 36.8%, 59.8% (P <0.05). 3. The effect on cell proliferation: MTT test assay showed that with pioglitazone concentration, pre-fat cells to an increasing trend, 100μmol/L pioglitazone significantly promote preadipocyte proliferation.In 24,48,72 hours ,the result of cell proliferation increased by 8.1%, 20.2% and 15.2% compared with control group (P<0.05).Conclusion: Adipocytokine as apelin can be secreted in preadipocyte, and its secretion is related with cell growth state. Pioglitazone can promote preadipocyte proliferation and differentiation, promote cell proliferation in time and dose dependent manner, its role in promoting differentiation in a time dependent manner. Thiazolidinedione can affect the secretion function of preadipocyte by regulating the state of cell growth.