Sun Huihui, Gao Chuanyuan, Yang Yumin, Liu Changqing, Qin Han, Tan Mengyuan, Li Jin, Li Xiaoxia, Du Kunze, Chang Yanxu
State Key Laboratory of Chinese Medicine Modernization, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
Tianjin Key Laboratory of Phytochemistry and Pharmaceutical Analysis, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
Mater Today Bio. 2025 Jun 23;33:102018. doi: 10.1016/j.mtbio.2025.102018. eCollection 2025 Aug.
The development of efficient methods for sensing αlpha-glucosidase (α-Glu) and screening its inhibitors has attracted significant attention due to their pivotal role in discovering therapeutic medicines for Type 2 diabetes. Herein, a low-cost and sensitive fluorometric strategy based on red carbon dots (R-CDs) and cobalt oxyhydroxide nanosheets (CoOOH NSs) had been established to detect α-Glu and screen its inhibitory compounds in natural products. As a switched fluorescence source, the fluorescence of R-CDs at 625 nm could be quenched by CoOOH NSs via Förster resonance energy transfer (FRET), assembled into nonfluorescent R-CDs@CoOOH nanocompositecomposites (R-CDs@CoOOH NCs). α-Glu hydrolyzed L-ascorbic acid-2---D-glucopyranose to produce ascorbic acid, which could reduce CoOOH NSs to Co, destroying R-CDs@CoOOH NCs and restoring the emission of red fluorescence. The proposed method exhibited a linear α-Glu range from 0.01 to 15 U mL and a low limit of detection (LOD) of 0.0037 U mL. Meanwhile, high-performance liquid chromatography-DAD-fraction collector (HPLC-DAD-FC) had been employed and combined with ultra-high-performance liquid chromatography-triple quadrupole time-of-flight mass spectrometry to isolate, enrich, and characterize compounds from (). This strategy was further extended by integrating the fluorometric platform with the HPLC-DAD-FC system to explore the inhibitory effects of extracts and anti-diabetic ingredients. Finally, 85 constituents were identified, with seven compounds exhibiting high α-Glu inhibitory activity. Consequently, the established strategy could accurately determine α-Glu in vitro and screen its inhibitors from natural products.
由于α-葡萄糖苷酶(α-Glu)在发现2型糖尿病治疗药物中起着关键作用,因此开发高效的α-葡萄糖苷酶传感方法及其抑制剂筛选方法备受关注。在此,基于红色碳点(R-CDs)和氢氧化钴纳米片(CoOOH NSs)建立了一种低成本、高灵敏度的荧光检测策略,用于检测α-Glu并筛选天然产物中的α-Glu抑制化合物。作为一种开关荧光源,R-CDs在625 nm处的荧光可通过Förster共振能量转移(FRET)被CoOOH NSs猝灭,组装成非荧光的R-CDs@CoOOH纳米复合材料(R-CDs@CoOOH NCs)。α-Glu将L-抗坏血酸-2---D-吡喃葡萄糖水解生成抗坏血酸,抗坏血酸可将CoOOH NSs还原为Co,破坏R-CDs@CoOOH NCs并恢复红色荧光发射。该方法检测α-Glu的线性范围为0.01至15 U/mL,检测限低至0.0037 U/mL。同时,采用高效液相色谱-二极管阵列检测器-馏分收集器(HPLC-DAD-FC)并结合超高效液相色谱-三重四极杆飞行时间质谱对()中的化合物进行分离、富集和表征。通过将荧光检测平台与HPLC-DAD-FC系统相结合,进一步拓展了该策略,以探索提取物和抗糖尿病成分的抑制作用。最终,鉴定出85种成分,其中7种化合物表现出较高的α-Glu抑制活性。因此,所建立的策略能够在体外准确测定α-Glu,并从天然产物中筛选其抑制剂。