To determine the effects of POCU1b on AMPK activation in 3 T3 L1 adipocytes, cells were treated with or without POCU1b and AMPK was de tected by immunoblot analysis. Levels of pAMPK were markedly increased protein inhibitor in a dose dependent manner following POCU1b treatment. At a same dose, these resulted in a 78% increase for POCU1b and 17% increase for HCA. Effect of POCU1b on expression of lipid accumulation related factors ADRP stimulates lipid accumulation and lipid drop let formation. Therefore, we tested the effects of POCU1b on the expression of ADRP, a lipid associated protein that is expressed during early adipose differenti ation. POCU1b reduced protein and mRNA expression levels of ADRP in a dose dependent manner. Similar to ADRP, perilipin stimulated lipid accumulation and localized to the surfaces of lipid droplets.
As shown Inhibitors,Modulators,Libraries in Figure 4, perilipin protein and mRNA expression levels decreased in POCU1b treated 3 T3 L1 adipocytes in a dose dependent manner. Effect of POCU1b on mRNA and protein expression levels of adipogenic factors To determine whether suppression of both triglycer ide accumulation and GPDH activity was caused by the inhibition of an adipogenic mechanism, we exam ined the expression levels of mRNA and protein of the adipogenic transcription factors PPAR and C EBP in POCU1b treated 3 T3 L1 adipocytes. As shown in Figure 5, POCU1b reduced both protein and mRNA expression levels of PPAR compared with standard differentiated adipocytes in a dose dependent manner. Both protein Inhibitors,Modulators,Libraries and mRNA expression levels of CEBP were also decreased in a dose dependent manner in POCU1b treated 3 T3 L1 cells.
Thus, we conclude that POCU1b Inhibitors,Modulators,Libraries regulates the production of adipogenic Inhibitors,Modulators,Libraries tran scription factors during the adipocyte differentiation process Inhibitors,Modulators,Libraries in 3 T3 L1 adipocytes. Discussion In the present study, POCU1b exhibited the strongest inhibitory effect on lipase activity and the putative anti obesity effects of POCU1b were investigated for the first time through the inhibition of adipocyte differentiation. POCU1b suppressed triglyceride accumulation in 3 T3 L1 adipocytes. GPDH activity was also significantly diminished by POCU1b in a dose dependent manner, and pAMPK activ ity was markedly increased in 3 T3 L1 adipocytes. Fur thermore, both ADRP and perilipin levels were decreased, and levels of the adipogenic transcription factors PPAR and CEBP were also significantly decreased after POCU1b treatment.
We also observed reduced lipid content in adipocytes, increased phosphorylation of AMPK, inhibition of lipid accumulation via lower ADRP and perilipin levels, and the inhibition of differentiation small molecule in 3 T3 L1 adipocytes. Natural products have been commonly used as dietary supplements to control body weight. P. cuspida tum has also been used as a natural laxative in trad itional Asian herbal medicines. In our study, the ethanol extract of P. cuspidatum exhibited strong anti lipase activity, with an IC50 value of 72. 5 67 ugml.