14-3-3γ Differentially Expressed In The WAT And Study The Role Of The Identified Proteins In Adipocyte Differentiation.

Objective: The aim of the present study was to uncover proteins -Pin1 and 14-3-3γ- differentially expressed in the WAT and study the role of the identified proteins in adipocyte differentiation.Methods and materials:1.In vivo:①Mouse model: 50 4-week-old male C57BL/6J mice were fed adaptability 7 days, and then were randomization into high fat diet group and low fat diet group, each group has 25 mice. The week after grouped was recorded as 0 week. The mice in the two groups were fed with high fat diet and low fat diet respectively for 14 weeks respectively. White adipose tissue was collected at 0, 4th and 14th week.②? Quantitative real-time PCR and western-blot analysis the expression of Pin1 and 14-3-3γin WAT of HFD and LFD group respectively.2.In vitro: During the differentiation of 3T3-L1 cells into adipocytes, quantitative real-time PCR and western-blot analysis the expression of 14-3-3γat d 0, d 2, d 4, d 6, d 8, d 10, d 12.3. In vitro: siRNA induced gene silencing of 14-3-3γ(mock as control) in adipocyte.?Oil red staining at d 8 which is the day preadipocyte turned into adipocyte. Quantitative real-time PCR analysis the expression of PPARγand C/EBPαat d 8 and d 6 respectively.Results:1.Consisted with the results of proteomics analysis, Pin1 was expressed lower at 4th week in WAT of HFD group while 14-3-3γexpressed higher.2. During the differentiation of 3T3-L1 cells into adipocytes, 14-3-3γexpressed higher at d 8. 3. Compared with mock, the?numbers of lipid droplet were fewer and the volume of lipid droplet was shrinking in adipocyte after siRNA induced gene silencing of 14-3-3γ.4. Compared with mock, the expression levels of PPARγand C/EBPαin adipocyte at d 8 and d 6 was lower.Conclusion:14-3-3γinhibit the adipocyte differentiation through the inhibition of the expression levels of PPARγand C/EBPαin adipocyte. We hope it will be target spot of the therapy of obesity.