Study On Function Of Ppp1r3c And NSDHL Gene During 3T3-L1 Preadipocytes Differentiation

Adipose tissue(adipose tissue)originated from the embryonic mesoderm,contain a variety of cells,the study of adipose tissue and its various cell types provides a good understanding for growth regulation,metabolism,endocrine function of adipose tissue and fat stem cell fate decision.Adipose tissue and fat cells have been one of the important areas that have been extensively explored by researchers and have important implications for identifying proteins or pathways that might be suitable targets for drug interventions.Analysis of RNA-Seq data showed that the expression of Protein phosphatase 1regulatory subunit 3c(Ppp1r3c)gene was significantly up-regulated and the NAD(P)steroid dehydrogenase like Protein(NSDHL)gene was significantly down-regulated in the adipogenic differentiation of adipocytes before 3T3-L1.This suggests that Ppp1r3 c and NSDHL may be new regulators in adipocyte differentiation,but their role in adipogenesis has not been reported,so we studied its role in adipogenesis.We report here that Ppp1r3 c small interfering RNA(si RNA)knocked out the expression of Ppp1r3 c,leading to the inhibition of fat production and fat gene expression.In contrast,the overexpression of Ppp1r3 c promoted the expression of fat-specific genes and the formation of lipid droplets in 3T3-L1 differentiated adipocytes.These results suggest that Ppp1r3 c may play a positive regulatory role in the formation of 3T3-L1 fat.In addition,we also found that Ppp1r3 c was down-regulated in 3T3-L1 interfered by SREBP1,suggesting that the influence of Ppp1r3 c on fat production may be realized through the regulation of SREBP1 in 3T3-L1.Previous RNA-Seq analyses revealed that NSDHL gene has a different expression during 3T3-L1 differentiation;however,its roles in adipogenesis are unknown.In the present study,using quantitative real-time PCR,we confirmed that NSDHL knockdown increased the proliferation of 3T3-L1 preadipocytes,but attenuated the differentiation of3T3-L1 preadipocytes,as evidenced by reduced lipid accumulation and downregulation of PPAR? gene expression.Further analyses showed that the expression peak of NSDHL was at the early stage of 3T3-L1 preadipocytes differentiation and LXR-SREBP1 signaling pathway was downregulated in NSDHL-knockdown 3T3-L1 cells.Collectively,our findings indicate that NSDHL is a novel modulator of adipogenesis.Moreover,our data provide insight into the complex relationships between sterol sensing,LXR-SREBP1 signaling pathway,and PPAR? in 3T3-L1 cells.In a word,this study shows that Ppp1r3 c and NSDHL are new regulators in the process of adipocyte differentiation,which provides a solid theoretical basis for future research on the molecular mechanism of adipogenesis.