## 4 Discussion  
Obesity is caused by an imbalance between energy intake and expenditure. Excess energy is stored in white adipose tissue in the form of triglycerides, and as cells grow and expand, excess fat accumulates [30]. A current research trend is using natural dietary compounds to treat and prevent obesity through fat browning and its associated lipid  
Figure 7: Effects of ESETM on thermogenesis and browning proteins expression in ICR mice fed HFD after 7 weeks and adipocytes treated with ESETM. (**a**) Western blots of thermogenesis and browning proteins in WAT. (**b**) Western blots of thermogenesis and browning proteins in adipocytes. Data are shown as the means \(\pm\) standard deviation (SD) (\(n\) = 5). Different low case letters above columns indicate statistical differences at \(p\) < 0.05.  
metabolism in energy expenditure [31]. The present study assessed whether ESETM could prevent obesity, inhibit fat accumulation, and upregulate proteins associated with lipolysis and browning pathways using high-fat-induced obese mice and 3T3 cells. Our findings will support ESETM as a novel anti-obesity agent with energy expenditure modulating properties. _E. stolonifera_, brown algae, is a natural functional raw material that can be used as a food material. Seaweeds are rich in various bioactive compounds, notably phlorotannin, which may have potential health benefits [32; 33; 34; 35; 36]. In 3T3-L1 adipocytes, _E. stolonifera_ inhibits adipogenesis activity by downregulating C/EBP\(\alpha\) and PPAR\(\gamma\) expression and improving lipolysis and adipocyte browning [23; 37]. However, the effects of clinical trial material made with ESE (ESETM) on adipocyte lipolysis and browning, on the other hand, have not been studied. Furthermore, more animal studies are required to assess the efficacy of ESETM as an anti-obesity agent. As a result, we investigated the anti-obesity effect of ESETM, using in vivo and in vitro models in this study. Excessive consumption of a high-fat diet may result in weight gain secondary to WAT accumulation [38]. As expected, mice fed the high-fat diet became obese and had higher WAT than mice fed the control diet. However, giving ESETM to HFD-fed mice for seven weeks significantly improved weight gain and decreased adipocyte size in WAT. HFD-induced obesity was also associated with higher serum triglyceride and cholesterol concentrations. In the present study, ESETM decreased serum TG, TC, and LDL-C and increased HDL-C levels [39]. We also measured adipogenic protein expression in WAT for each group using Western blot analysis. ESETM reduced the expression of C/EBP\(\alpha\), PPAR\(\gamma\), and aP2 in HFD-fed mice, which are required for the late stages of preadipocyte differentiation. Furthermore, ESETM reduced the expression of the lipogenic proteins SREBP1, LPAAT0, and lipin1, which may reduce lipid synthesis. These proteins' expression was also reduced to a similar extent after treatment of 3T3-L1 cells with ESETM. Thus, in vivo and in vitro data suggest that ESETM can prevent the development of HFD-induced obesity by regulating adipogenesis and the expression of adipogenesis factors. Lipolysis is the hydrolysis of triglycerides stored in adipocytes. This hydrolysis process is activated by PKA and induces phosphorylation of HSL to release free fatty acids for oxidation in mitochondria. PKA can also induce lipolytic enzymes, such as ATGL, HSL, and MGL [40; 41; 42]. In this study, we prove that ESETM treatment increased the expression of p-PKA, ATGL, p-HSL, and MGL in HFD-fed mice. Expressed in the inner mitochondrial membrane, UCP1 is a hallmark brown adipocyte protein responsible for the energy-expending phenotype of brown adipose tissue. Furthermore, increased UCP1 expression suggests that mitochondrial activity is critical for the browning effect, and its expression in WAT drives the development of brown adipose tissue-like features [43; 44].