Background: The elucidation of molecular pathways associated with adipogenesis has evidenced the relevance of estrogen and estrogen receptor beta (ERβ). The positive effects of ERβ ligands on adipogenesis, energy expenditure, lipolysis, food intake, and weight loss, make ERβ an attractive target for obesity control. From ligand-based virtual screening, molecular docking, and molecular dynamic simulations, six new likely ERβ ligands (C1 to C6) have been reported with potential for pharmacological obesity treatment. Objective: In this study, the effect of molecules C1-C6 on adipogenesis using the murine 3T3-L1 cell line was evaluated. Methods: Cell viability was assessed by MTT assays. Lipid accumulation and gene expression were investigated by ORO staining and real-time quantitative RT-PCR experiments, respectively. Results: Cell viability was not significantly affected by C1-C6 at concentrations up to 10 μM. Interestingly, treatment with 10 μM of C1 (S-Dihydrodaidzein) and C2 (3-(1,3-benzoxazol-2-yl)- benzamide) for 72 h inhibited adipocyte differentiation; moreover, ORO staining evidenced a reduced intracellular lipid accumulation (40% at day 7). Consistently, mRNA expression of the adipogenic markers, PPARγ and C/EBPα, was reduced by 50% and 82%, respectively, in the case of C1, and by 83% and 59%, in the case of C2. Conclusion: Altogether, these results show the two new potential β-estrogen receptor ligands, C1 and C2, to exhibit anti-adipogenic activity. They could further be used as lead structures for the development of more efficient drugs for obesity control.

中文翻译：

---

S-二氢黄豆苷元和3-(1,3-苯并恶唑-2-基)-苯甲酰胺，两种具有抗脂肪活性的新的潜在β-雌激素受体配体


背景：与脂肪生成相关的分子途径的阐明已经证明了雌激素和雌激素受体β（ERβ）的相关性。 ERβ 配体对脂肪生成、能量消耗、脂肪分解、食物摄入和体重减轻的积极作用，使 ERβ 成为肥胖控制的有吸引力的靶点。通过基于配体的虚拟筛选、分子对接和分子动力学模拟，已报道了六种新的可能的 ERβ 配体（C1 至 C6），它们具有药物肥胖治疗的潜力。目的：在本研究中，使用鼠 3T3-L1 细胞系评估分子 C1-C6 对脂肪形成的影响。方法：通过 MTT 测定评估细胞活力。分别通过ORO染色和实时定量RT-PCR实验研究脂质积累和基因表达。结果：浓度高达 10 μM 的 C1-C6 并未显着影响细胞活力。有趣的是，用10μM的C1（S-二氢黄豆苷元）和C2（3-（1,3-苯并恶唑-2-基）-苯甲酰胺）处理72小时抑制脂肪细胞分化；此外，ORO 染色证明细胞内脂质积累减少（第 7 天为 40%）。一致的是，在 C1 中，脂肪形成标志物 PPARγ 和 C/EBPα 的 mRNA 表达分别降低了 50% 和 82%，在 C2 中分别降低了 83% 和 59%。结论：总而言之，这些结果表明两种新的潜在 β-雌激素受体配体 C1 和 C2 具有抗脂肪形成活性。它们可以进一步用作开发更有效的肥胖控制药物的先导结构。