PDF(Pubmed)
Abstract：
Pueraria lobata (P. lobata), a traditional anti-diabetic medicine mainly composed of flavonoids and isoflavones, has a long history in diabetes treatment in China. However, the anti-diabetic active component is still unclear. Recently, protein tyrosine phosphatase 1B (PTP1B) has been a hot therapeutic target by negatively regulating insulin signaling pathways. In this study, the spectrum-effect relationship analysis method was first used to identify the active components of P. lobata that inhibit PTP1B. The fingerprints of 12 batches of samples were established using high-performance liquid chromatography (HPLC), and sixty common peaks were identified. Meanwhile, twelve components were identified by a comparison with the standards. The inhibition of PTP1B activity was studied in vitro by using the p-nitrophenol method, and the partial least squares discriminant analysis, grey relational analysis, bivariate correlation analysis, and cluster analysis were used to analyze the bioactive compounds in P. lobata. Peaks 6, 9 (glycitin), 11 (genistin), 12 (4\'-methoxypuerarin), 25, 34, 35, 36, 53, and 59 were considered as potentially active substances that inhibit PTP1B. The in vitro PTP1B inhibitory activity was confirmed by glycitin, genistin, and 4\'-methoxypuerarin. The IC50s of the three compounds were 10.56 ± 0.42 μg/mL, 16.46 ± 0.29 μg/mL, and 9.336 ± 0.56 μg/mL, respectively, indicating the obvious PTP1B inhibitory activity. In brief, we established an effective method to identify PTP1B enzyme inhibitors in P. lobata, which is helpful in clarifying the material basis of P. lobata on diabetes. Additionally, it is evident that the spectrum-effect relationship method serves as an efficient approach for identifying active compounds, and this study can also serve as a reference for screening bioactive constituents in traditional Chinese medicine.
摘要：
葛根(P.lobata),一种传统的抗糖尿病药物，主要由黄酮类和异黄酮组成，糖尿病治疗在中国有着悠久的历史。然而,抗糖尿病活性成分尚不清楚.最近,蛋白酪氨酸磷酸酶1B(PTP1B)通过负向调节胰岛素信号通路成为热门的治疗靶点。在这项研究中,首先采用谱效关系分析方法鉴定了抑制PTP1B的P.bobata活性成分。采用高效液相色谱法(HPLC)建立了12批样品的指纹图谱,并确定了60个常见峰。同时,通过与标准品的比较,确定了12种成分.通过使用对硝基苯酚方法在体外研究了PTP1B活性的抑制作用，和偏最小二乘判别分析，灰色关联分析，双变量相关分析，并采用聚类分析对P.lobata中的生物活性化合物进行分析。峰6，9（缩水甘油），11（生苷），12（4'-甲氧基葛根素），25、34、35、36、53和59被认为是抑制PTP1B的潜在活性物质。体外PTP1B抑制活性由缩水甘油证实，Genistin,和4'-甲氧基葛根素。三种化合物的IC50为10.56±0.42μg/mL,16.46±0.29μg/mL，和9.336±0.56μg/mL，分别,表明明显的PTP1B抑制活性。简而言之,我们建立了一种有效的方法来鉴定假单胞菌中的PTP1B酶抑制剂，这有助于阐明假单胞菌对糖尿病的物质基础。此外,显然，光谱-效应关系方法是识别活性化合物的有效方法，本研究也可为中药中生物活性成分的筛选提供参考。
