# Anti-Obesity Effect of Fresh and Browned _Magnolia denudata_ Flowers in 3T3-L1 Adipocytes
## 4 Discussion  
Processing tea fermentation is a natural post-oxidation process that involves incubating the withered-fresh tea leaves in a closed high or low-humidity storage chamber for a defined period. The chemical components of fermented tea are different from those of green tea. Many studies have shown fermented tea's antioxidant, anti-mutagenic, anti-atherosclerotic, anti-obesity, and anti-hypercholesterol effects [27; 28; 29; 30; 31]. In this study, the _Magnolia denudata_ flower was fermented like oolong and black tea. Several studies have reported on 3T3-L1 cells with fresh _Magnolia denudate_ flower [9; 10; 11; 12]. However, our study is different because we focused on the anti-obesity effects to observe the detailed molecular mechanism with fresh and browned magnolia flowers. Thus, study is the first of its kind. We identified the chemical components of fresh and browned _Magnolia denudata_ flower, and the change in the chemical components of _Magnolia denudata_ flower was expected to affect its physiological activities [1; 26]. This is because the chemical compounds of MFEs could be a basis for showing the potential for many biological activations. However, it needs to be mentioned that these chemical compounds are cytotoxic if used in a high concentration, as shown in Figure 1, which demonstrates that cell viability decreases with an increasing concentration of more than 1000 \(\mathrm{\SIUnitSymbolMicro g}\)/mL. Based on this finding, we selected much safer concentrations of less than 500 \(\mathrm{\SIUnitSymbolMicro g}\)/mL, such as 200 and 400 \(\mathrm{\SIUnitSymbolMicro g}\)/mL. PARy (peroxisome proliferatorator-activating receptor gamma), C/EBP\(\alpha\) (CCAAT/inhancer-binding protein alpha), and SREBP-1 (cholesterol-regulated factor-binding protein 1) are major transcription factors that play an important role in adipogenic and lipogenic control. PPARy has a role as a major regulator of adipogenesis. It is essential for differentiating adipocyte into mature adipocytes. C/EBP\(\alpha\) interacts with PPARy to enhance the expression of the genes required for adipocyte differentiation. SREBP-1 plays a significant role in fat production. These transcription factors make a complex network that ensures the proper development and functioning of adipose tissue [32]. It has been demonstrated that the expression inhibition of PPARy, C/EBP\(\alpha\), and SREBP-1 in adipocytes is associated with the prevention of obesity [33; 34]. This is because the transcription factors regulate the gene expression of adipogenic targets in adipocytes. The FAS (fatty acid synthase) uses acetyl-CoA and malonyl-CoA as a substrate to generate long-chain fatty acids and promote lipogenesis [35; 36; 37]. Perilipin participates in the storage and mobilization of lipids [38; 39; 40; 41; 42]. FABP4 (Fatty Acid-Binding Protein 4) is a carrier protein for fatty acids in adipocytes and  
Figure 5: Effect of FMFE and BMFE on the energy and lipid metabolism in 3T3-L1 adipocytes. (**A**) p-ACC, (**B**) p-AMPK. Different letters indicate significant differences at \(p<0.05\). is responsible for intracellular lipid absorption and storage development [43; 44; 45]. HSL (hormone-sensitive lipase) regulates the triacylglycerol hydrolysis into free fatty acids in lipolysis metabolism [46; 47; 48]. In the present study, we found that the mRNA expression of all adipogenic target genes significantly decreased by treatment with FMFE and BMFE. We also found that the protein expression of adipogenic target proteins such as PPAR\(\gamma\), C/EBP\(\alpha\), and perilipin significantly decreased by treatment with FMFE and BMFE. These findings suggest that treatment with FMFE and BMFE induces the inhibition of PPAR\(\gamma\), C/EBP\(\alpha\), SREBP-1, FAS, perilipin, FABP4, and HSL in adipocytes. Cells need to act favorably to available nutrients by amending their metabolism for their energy needs. AMPK (AMP-activated protein kinase) is a highly conserved serine/threonine energy-sensing kinase.