Abstract：
Targeting multiple factors such as oxidative stress, alpha glucosidase and acetylcholinesterase (AChE) are considered advantageous for the treatment of diabetes and diabetes associated-cognitive dysfunction. In the present study, Hibiscus rosa-sinensis flowers anthocyanin-rich extract (HRA) was prepared. Phytochemical analysis of HRA using LC-ESI/MS/MS revealed the presence of various phenolic acids, flavonoids and anthocyanins. HRA showed in vitro antioxidant activity at low concentrations. HRA inhibited all the activities of mammalian glucosidases and AChE activity. The IC50 value of HRA for the inhibition of maltase, sucrase, isomaltase, glucoamylase and AChE was found to be 308.02 ± 34.25 µg/ml, 287.8 ± 19.49 µg/ml, 424.58 ± 34.75 µg/ml, 408.94 ± 64.82 µg/ml and 264.13 ± 30.84 µg/ml, respectively. Kinetic analysis revealed mixed-type inhibition against all the activities except for glucoamylase (competitive) activity. In silico analysis confirmed the interaction of two active constituents cyanidin 3-sophoroside (CS) and quercetin 3-O-sophoroside (QS) with four subunits, n-terminal and c-terminal subunits of human maltase-glucoamylase and sucrase-isomaltase as well as with AChE. Molecular dynamics simulation, binding free energy calculation, DCCM, PCA, PCA-based free energy surface analysis ascertained the stable binding of CS and QS with target proteins studied. HRA could be used as complementary therapy for diabetes and cognitive improvement.
摘要：
针对多种因素，如氧化应激，α-葡糖苷酶和乙酰胆碱酯酶(AChE)被认为有利于糖尿病和糖尿病相关认知功能障碍的治疗。在本研究中,制备了富含花青素的芙蓉花提取物(HRA)。使用LC-ESI/MS/MS对HRA进行植物化学分析，发现存在各种酚酸，黄酮类和花青素。HRA在低浓度下显示出体外抗氧化活性。HRA抑制哺乳动物葡萄糖苷酶的所有活性和AChE活性。HRA对麦芽糖酶抑制作用的IC50值，蔗糖酶,异麦芽糖酶，发现葡糖淀粉酶和AChE为308.02±34.25µg/ml，287.8±19.49微克/毫升，424.58±34.75µg/ml，408.94±64.82µg/ml和264.13±30.84µg/ml，分别。动力学分析显示，除葡糖淀粉酶（竞争性）活性外，对所有活性均具有混合型抑制作用。计算机模拟分析证实了两种活性成分花青素3-苦参苷（CS）和槲皮素3-O-苦参苷（QS）与四个亚基的相互作用，人麦芽糖-葡糖淀粉酶和蔗糖酶-异麦芽糖酶以及AChE的n端和c端亚基。分子动力学模拟,结合自由能计算，DCCM,PCA,基于PCA的自由能表面分析确定了CS和QS与所研究的靶蛋白的稳定结合。HRA可作为糖尿病和认知改善的补充治疗。
