Temviriyanukul Piya, Inthachat Woorawee, Jaiaree Ararat, Karinchai Jirarat, Buacheen Pensiri, Yodkeeree Supachai, Laowanitwattana Tanongsak, Chewonarin Teera, Suttisansanee Uthaiwan, Imsumran Arisa, Wongnoppavich Ariyaphong, Pitchakarn Pornsiri
Institute of Nutrition, Mahidol University, Salaya, Nakhon Pathom 73170, Thailand.
Department of Biochemistry, Faculty of Medicine, Chiang Mai University, Muang Chiang Mai, Chiang Mai 50200, Thailand.
Foods. 2025 Jul 24;14(15):2603. doi: 10.3390/foods14152603.
Thai fermented soybeans (TFSs) contain phytochemicals with anti-diabetic benefits. In this study, an initial non-optimized TFS extract (TFSE) was prepared using a conventional triplicate 80% ethanol extraction method and evaluated for its biological activity. TFSE effectively reversed TNF-α-induced insulin resistance in 3T3-L1 adipocytes by enhancing insulin-stimulated glucose uptake, indicating anti-diabetic potential. TFSE also upregulated the phosphorylation of AKT (a key insulin signaling mediator) and the expression of adipogenic proteins (PPARγ, CEBPα) in TNF-α-exposed 3T3-L1, suggesting the mitigation of adipocyte dysfunction; however, the results did not reach statistical significance. The conventional extraction process was labor-intensive and time-consuming, and to enhance extraction efficiency and bioactivity, the process was subsequently optimized using environmentally friendly microwave-assisted extraction (MAE) in combination with the Box-Behnken design (BBD) and response surface methodology (RSM). The optimized extract (O-TFSE) was obtained over a significantly shorter extraction time and exhibited higher levels of total flavonoids and antioxidant activity in comparison to TFSE, while showing reduced levels of isoflavones (daidzein, genistein, and glycitein) in relation to TFSE. Interestingly, O-TFSE retained similar efficacy in reversing TNF-α-induced insulin resistance and demonstrated significantly stronger α-glucosidase and α-amylase inhibitory activities, indicating its enhanced potential for diabetes management. These results support the use of MAE as an efficient method for extracting functional compounds from TFS for functional foods targeting insulin resistance and type 2 diabetes mellitus.
泰国发酵大豆（TFSs）含有具有抗糖尿病益处的植物化学物质。在本研究中，使用传统的一式三份80%乙醇提取方法制备了初始未优化的TFS提取物（TFSE），并对其生物活性进行了评估。TFSE通过增强胰岛素刺激的葡萄糖摄取，有效逆转了TNF-α诱导的3T3-L1脂肪细胞中的胰岛素抵抗，表明其具有抗糖尿病潜力。TFSE还上调了TNF-α处理的3T3-L1中AKT（一种关键的胰岛素信号介质）的磷酸化以及脂肪生成蛋白（PPARγ、CEBPα）的表达，提示减轻了脂肪细胞功能障碍；然而，结果未达到统计学显著性。传统提取过程劳动强度大且耗时，为提高提取效率和生物活性，随后采用环境友好的微波辅助提取（MAE）结合Box-Behnken设计（BBD）和响应面方法（RSM）对该过程进行了优化。与TFSE相比，优化提取物（O-TFSE）在显著更短的提取时间内获得，总黄酮水平和抗氧化活性更高，而异黄酮（大豆苷元、染料木黄酮和黄豆黄素）水平相对于TFSE降低。有趣的是，O-TFSE在逆转TNF-α诱导的胰岛素抵抗方面保持了相似的功效，并表现出显著更强的α-葡萄糖苷酶和α-淀粉酶抑制活性，表明其在糖尿病管理方面具有增强的潜力。这些结果支持将MAE作为从TFS中提取功能性化合物的有效方法，用于针对胰岛素抵抗和2型糖尿病的功能性食品。
