## 4 Discussion  
Obesity, which is characterized by overweight, is heavily influenced by changing social, economic, lifestyle, and nutritional conditions, and populations of all ages, including adults, the elderly, infants, and adolescents, are at risk of developing obesity-related diseases [26; 27]. In addition, a society that focuses on physical appearance has naturally led to an interest in dietary control and functional health foods for weight loss, and research on supplements that help control weight to meet these needs is also actively being conducted [28; 29; 30]. IG, which is rich in vitamin C and minerals, has long been used as a herbal medicine in ancient Indian Ayurvedic medicine and has recently been found to contain active substances, such as gallic acid and ellagic acid [12; 13]. BP, which is rich in dietary fiber and antioxidants, is a cultivated plant in many parts of the world and has recently been identified to possess active substances, such as saponarin and lutonarin [18; 19; 20]. IG and BP have been reported to affect lipid metabolism individually, but their synergistic effects are unknown. Our study aimed to determine whether an IGBP mixture exerts effects on the adipogenesis and lipogenesis mechanisms of differentiated 3T3-L1 cells. In a previous study, we confirmed that the IGBP mixture showed a lipolytic effect on mature adipocytes, and the cell viability experiment also showed no toxicity up to the maximum concentration (200 \(\mu\)g/mL) used in this experiment. Regarding the mixing ratio of the IGBP mixture, the ratios that were most effective in the ratio determination test conducted before the  
Figure 5: Effects of the IGBP 2:1 mixture on lipid accumulation, triglyceride (TG) levels, and glycerol release in 3T3-L1 adipocytes: (**A**) Oil Red O staining, (**B**) optical density of lipid accumulation, (**C**) TG levels, and (**D**) glycerol release. NC, normal control; C, differentiated control; Met, metformin 1 mM; IG200, IG 200 \(\mu\)g/mL; BP200, BP 200 \(\mu\)g/mL; IGBP 2:1 (50, 100, or 200 \(\mu\)g/mL). The resulting values are shown as mean \(\pm\) standard deviation (SD; \(n=3\)), and the different superscript letters (alphabet) indicate significance at \(p<0.05\). start of this experiment were selected first, and the 2:1 (IG:BP) ratio was ultimately selected to account for the available supply of raw material. To clarify the effect of the IGBP 2:1 mixture on the adipogenesis and lipogenesis mechanisms in mature adipocytes, we measured the expression and activity levels of adipogenesis- and lipogenesis-related proteins in 3T3-L1 adipocytes. Adipogenesis is the process by which the differentiation of pre-adipocytes to mature adipocytes takes place [31]. It begins with a decrease in the cAMP level. This increases the activation of CREB and affects the expression of transcription factors, such as PPARy and C/EBP\(\alpha\)[32]. Adipogenesis is regulated by several signaling pathways or transcription factors that are master regulators in the final stage of differentiation [33]. PPARy, a ligand-activated transcription factor, can induce the expression of C/EBP\(\alpha\), which subsequently mediates the expression levels of adipogenesis-related genes, including FABP4, adiponectin, leptin, and G6 Pase [9; 10]. In the present study, the expression levels of cAMP and MAPK in adipogenic-induced adipocytes decreased. Although the phosphorylation of MAPK has not been confirmed, it is assumed that MAPK was phosphorylated and converted to an active form. In addition, it was confirmed that the expression levels of C/EBP\(\beta\), C/EBP\(\alpha\), and PPARy increased in the adipocyte differentiation model. Thus, the IGBP 2:1 mixture increased the cAMP expression levels compared to the differentiation-inducing group, reduced the activity of CREB, and decreased the expression levels of C/EBP\(\beta\), C/EBP\(\alpha\), and PPARy.