## 4 Discussion
LPL encodes a lipoprotein lipase expressed in the heart, muscle tissue, and adipose tissue [7; 40]. Additionally, it plays an important role in breaking down the triglyceride-type fats into fats, and the broken fat molecules are used as energy in the body or stored in adipose tissue [41; 42; 43]. In this study, the expression of LPL in 3T3-L1 cells was confirmed by RT-qPCR and Western blot analysis (Figure 6). Similar to that reported by [PERSON] et al. [44], the expression of LPL was decreased in a dose-dependent manner in the 0.025 and 0.05 mg/mL ABF(r) groups compared to the ABF(r) untreated group. Lipid accumulation and adipocyte differentiation are major signals in the development of obesity [15; 16]. Transcription factors that induce gene expression in the adipogenic pathway can differentiate pre-adipocytes into mature adipocytes [9; 10]. The expression of C/EBP\(\beta\) is induced in the early stages of 3T3-L1 differentiation and regulates the expression of PPAR\(\gamma\) and C/EBP\(\alpha\), which are major transcription factors of adipogenesis and adipogenesis [36; 45]. The adipogenic differentiation process is regulated by complex and systematic gene expression, in which CCAAT enhancer-binding protein alpha (C/EBP\(\alpha\)) and peroxisome proliferator-activated receptor-gamma (PPAR\(\gamma\)) are two major regulators. FABPs are low molecular weight (14-15 kDa) proteins that occur in various tissues. FABP plays an important role in fatty acid metabolism and transport [39; 46; 47]. FABP2 is known to regulate fatty acids and cholesterol during adipocyte differentiation [48; 49]. FABP2 plays an important role in several long-chain fatty acid transport steps and is associated with increased intestinal fat absorption and obesity [48; 50; 51]. In the final step, FAS regulates adipocyte differentiation [6; 52; 53]. Therefore, we examined the effects of ABF(r) on the mRNA and protein expression at adipogenesis levels of C/EBP\(\alpha\), PPAR\(\gamma\), FABP2, and FAS in 3T3-L1 adipocytes (Figure 7). Similar to the results of a previous study [37; 38; 54; 55], ABF(r) inhibited the expression of C/EBP\(\alpha\), PPAR\(\gamma\), FABP2, and FAS in adipocytes. Thus, it could prevent obesity by blocking early adipocyte differentiation. Adipose tissue is to express and secrete various peptides, known as adipokines, and express several receptors that can respond to other signals from the hormone system and the central nervous system [5]. Adipose tissue is metabolically active and regulated by energy expenditure through both \"nervous and endocrine factors\" [56]. Adipokines play an important role in obesity-induced pathophysiological changes at the dysfunctional adipose tissue and cardiometabolic alterations [52; 56]. Adiponectin regulates lipid storage and adipogenesis in 3T3-L1 cells [57]. Leptin is produced in adipose tissue and plays an important role in body fat storage associated with food intake, energy homeostasis, and other physiological processes [14; 28]. We assessed the role of ABF(r) in adipokine expression during adipogenesis in 3T3-L1 cells by ELISA (Figure 8). As in the previous study by [PERSON] et al. [58], ABF(r) treatment of differentiated 3T3-L1 cells significantly enhanced adiponectin secretion in adipocytes and decreased leptin secretion in differentiated 3T3-L1 cells in a dose-dependent manner. These results show that ABF(r) has anti-adipogenic potential in 3T3-L1 cells. However, the present study has limitations that should be noted. Firstly, the effects of ABF(r) on protein or gene expressions associated with lipolysis, such as adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), were not examined. As the inhibition of differentiation, as well as inductions of lipolysis, has been focused on therapeutic strategies for treating obesity [59], further studies are necessary to investigate the effects of ABF(r) on lipolysis, another important pathway for suppressing fat accumulation. Secondly, our results were obtained in vitro only.