Translational Medicine Research Center, Guizhou Medical University, Guiyang, Guizhou 550025, China; Institute of Interdisciplinary Integrative Medicine Research, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China.
Guizhou Provincial Key Laboratory of Pathogenesis and Drug Research on Common Chronic Diseases, Guizhou Medical University, Guiyang, Guizhou 550025, China.
J Pharm Biomed Anal. 2024 Oct 15;249:116382. doi: 10.1016/j.jpba.2024.116382. Epub 2024 Jul 26.
DPP-IV inhibitors, which are close to the natural hypoglycemic pathway of human physiology and have few side effects, have been extensively employed in the management of type 2 diabetes mellitus (T2DM). However, there are currently no specific blood indicators that can indicate or predict a patient's suitability for DPP-IV inhibitors. In this study, based on the self-developed high-specificity fluorescent substrate glycyl-prolyl-N-butyl-4-amino-1, 8-naphthimide (GP-BAN), a detection method of human serum DPP-IV activity was established and optimized. The method demonstrates a favorable lower limit of detection (LOD) at 0.32 ng/mL and a satisfactory lower limit of quantification (LOQ) of 1.12 ng/mL, and can be used for the detection of DPP-IV activity in trace serum (2 μL). In addition, Vitalliptin and Sitagliptin showed similar IC values when human recombinant DPP-IV and human serum were used as enzyme sources, and the intra-day and inter-day precision obtained by the microplate analyzer were less than 15 %. These results indicate that the microplate reader based detection technique has good accuracy, repeatability and reproducibility. A total of 700 volunteers were recruited, and 646 serum samples were tested for DPP-IV activity. The results showed that serum DPP-IV activity was higher in patients with T2DM than in controls (P < 0.01). However, the statistical data of family history of diabetes, gender and age of diabetic patients showed no statistical significance, and there was no contrast difference. The DPP-IV activity of serum in T2DM patients ranged from 2.4 μmol/min/L to 78.6 μmol/min/L, with a huge difference of up to 32-fold. These results suggest that it is necessary to test DPP-IV activity in patients with T2DM when taking DPP-IV inhibitors to determine the applicability of DPP-IV inhibitors in T2DM patients. These results suggest that it is necessary to detect the activity of DPP-IV in blood before taking DPP-IV inhibitors in patients with T2DM to judge the applicability of DPP-IV inhibitors in patients with T2DM.
二肽基肽酶-4（DPP-4）抑制剂与人体生理的天然降糖途径较为接近，且副作用较少，因此被广泛用于 2 型糖尿病（T2DM）的治疗。然而，目前尚无特定的血液指标可以指示或预测患者是否适合使用 DPP-4 抑制剂。在本研究中，我们基于自主研发的高特异性荧光底物甘氨酰-脯氨酰-N-丁基-4-氨基-1,8-萘啶（GP-BAN），建立并优化了一种检测人血清 DPP-4 活性的方法。该方法的检测下限（LOD）为 0.32ng/mL，定量下限（LOQ）为 1.12ng/mL，检测痕量血清（2μL）中的 DPP-4 活性时，结果令人满意。此外，以人重组 DPP-4 和人血清为酶源时，维格列汀和西格列汀的 IC 值相似，微板分析仪获得的日内和日间精密度均小于 15%。这些结果表明，基于微板阅读器的检测技术具有良好的准确性、重复性和再现性。我们共招募了 700 名志愿者，检测了 646 份血清样本的 DPP-4 活性。结果显示，T2DM 患者的血清 DPP-4 活性明显高于对照组（P＜0.01）。然而，糖尿病患者的家族史、性别和年龄的统计数据无统计学意义，无显著性差异。T2DM 患者的血清 DPP-4 活性范围为 2.4μmol/min/L 至 78.6μmol/min/L，差异高达 32 倍。这些结果提示，T2DM 患者在使用 DPP-4 抑制剂前应检测 DPP-4 活性，以确定 DPP-4 抑制剂在 T2DM 患者中的适用性。这些结果提示，在 T2DM 患者使用 DPP-4 抑制剂前应检测血液中的 DPP-4 活性，以判断 DPP-4 抑制剂在 T2DM 患者中的适用性。
