Wei Yong, Xuan Yuye, Wang Wenxiang, Zhang Yonghui, Li Xiaoshan, Liu Chunlan, Wang Chen, Liu Zhihai
College of Electronic and Information Engineering, Chongqing Three Gorges University, Chongqing 404100, China.
Chongqing Key Laboratory of Development and Utilization of Genuine Medicinal Materials in Three Gorges Reservoir Area, Chongqing Three Gorges Medical College, Chongqing 404120, China.
Biomed Opt Express. 2025 Feb 18;16(3):1090-1103. doi: 10.1364/BOE.555014. eCollection 2025 Mar 1.
The existing screening methods for therapeutic targets of active ingredients in traditional Chinese medicine (TCM) have problems of long detection time and high instrument cost. This article proposes a new target screening method based on virtual screening and fiber surface plasmon resonance (SPR) sensing technology, which has the characteristics of flexibility, speed, and low cost. It also reveals the target mechanism of the active ingredient isoquercitrin in the treatment of insulin resistance (IR). The binding energies of isoquercitrin with target proteins PDPK1, INSR, and PTPN1 were calculated using computer virtual methods to be -8.9, -8.9, -8.8 kcal/mol, indicating strong binding activity with isoquercitrin and predicted as three key targets. Then a fiber optic SPR biosensor functionalized with isoquercitrin molecules was constructed to detect the binding affinity between isoquercitrin and the key targets. The experimental results showed that the binding affinities of isoquercitrin to the targets PDPK1, INSR, and PTPN1 were 1.45, 1.14, and 13.21, respectively, indicating that PTPN1 is the main target of isoquercitrin in the treatment of IR. The proposed sensor has a sensitivity of 0.699 nm/(μg/ml), LOD of 0.515μg/ml, and the experimental detection time of this method is as low as 45 minutes, without the need for large and expensive optical demodulation equipment, and the device volume is 5.50 dm, providing new ideas for the screening of therapeutic targets of active ingredients in TCM.
现有中药活性成分治疗靶点的筛选方法存在检测时间长和仪器成本高的问题。本文提出了一种基于虚拟筛选和光纤表面等离子体共振（SPR）传感技术的新型靶点筛选方法，该方法具有灵活性、速度快和成本低的特点。同时揭示了活性成分异槲皮苷治疗胰岛素抵抗（IR）的靶点机制。采用计算机虚拟方法计算异槲皮苷与靶点蛋白PDPK1、INSR和PTPN1的结合能分别为-8.9、-8.9、-8.8 kcal/mol，表明与异槲皮苷具有较强的结合活性，并预测为三个关键靶点。然后构建了异槲皮苷分子功能化的光纤SPR生物传感器，检测异槲皮苷与关键靶点之间的结合亲和力。实验结果表明，异槲皮苷与靶点PDPK1、INSR和PTPN1的结合亲和力分别为1.45、1.14和13.21，表明PTPN1是异槲皮苷治疗IR的主要靶点。所提出的传感器灵敏度为0.699 nm/(μg/ml)，检测限为0.515μg/ml，该方法的实验检测时间低至45分钟，无需大型昂贵的光解调设备，装置体积为5.50 dm，为中药活性成分治疗靶点的筛选提供了新思路。
