Screening and functional evaluation of the glucose-lowering active compounds of total saponins of Baibiandou(Lablab Semen Album)  

白扁豆总皂苷降糖活性组分筛选及其功能评价

作　　者：HAN Jun ZHENG Qinfang FANG Liangzi HUANG Xiaolong 韩君;郑钦方;方良子;黄小龙(北京康仁堂药业有限公司中药配方颗粒关键技术国家地方联合工程研究中心,北京101301;湖南中医药大学科技创新中心,湖南长沙410208;湖南医药学院侗医药研究湖南省重点实验室,湖南怀化418000;无锡霍普生物科技有限公司分析测试中心,江苏无锡214000)

机构地区：[1]National and Local Joint Engineering Research Center for Key Technology of Chinese Medicinal Composition Granules,Beijing Tcmages Pharmaceutical Co.,Ltd.,Beijing 101301,China [2]Science and Technology Innovation Center,Hunan University of Chinese Medicine,Changsha,Hunan 410208,China [3]Key Laboratory of Dong Medical Research of Hunan Province,Hunan University of Medicine,Huaihua,Hunan 418000,China [4]Analysis and Testing Center of Hope Biotechnology Co.,Ltd.,Wuxi,Jiangsu 214000,China

出　　处：《Digital Chinese Medicine》2021年第3期229-240,共12页数字中医药（英文）

基　　金：funding support from the Program of the Educational Commission of Hunan Province of China(No.20B418)。

摘　　要：Objective To screen forα-glucosidase inhibitor active compounds in the total saponins of Baibiandou(Lablab Semen Album)based on UHPLC-Q-Exactive Orbitrap MS technology and to evaluate its hypoglycemic activity in vivo.Methods Acarbose was used as the positive control,and the median inhibitory concentration(IC50)was used as the evaluation index ofα-glucosidase inhibitory activity to establish an in vitroα-glucosidase inhibition model.Further,UHPLC-Q-Exactive Orbitrap MS technology was used to screen and identify the active compounds ofα-glucosidase inhibitors in the total saponins of Baibiandou(Lablab Semen Album)in order to further verify the activity of the main active monomer and to perform homologous modeling and molecular docking of yeast-derivedα-glucosidase and human-derivedα-glucosidase,while the hypoglycemic activity was evaluated in diabetic mice.Results This study successfully identified 15 compounds with potentialα-glucosidase inhibitory activity,including Chikusetsusaponin IVa,from the total saponins of Baibiandou(Lablab Semen Album).Simultaneously,we verified the activity of the main active monomer Chikusetsusaponin IVa,and showed that it has strongα-glucosidase inhibitory activity.Theα-glucosidase inhibitory concentration IC50 was(565.2±1.026)μg/m L,and the IC50 of acarbose,which was lower than the positive control,was(706.6±1.058)μg/m L.The docking energies of Chikusetsusaponin IVa were–6.1 and–7.7 kcal/mol with yeast-derivedα-glucosidase and human-derivedα-glucosidase molecules,respectively.Both showed strong binding activity,and the levels of alanine aminotransaminase(ALT),aspartate aminotransaminase(AST),UREA,Creatinine(CREA),and cholesterol(CHO)were significantly decreased by Chikusetsusaponin IVa(P<0.05).In addition,it could repair damaged liver and pancreas cells of diabetic mice to some extent.Conclusion This study provides a basis for screeningα-glucosidase inhibitors and structural modifications of the total saponins of Baibiandou(Lablab Semen Album).目的基于UHPLC-Q-Exactive Orbitrap MS技术筛选白扁豆总皂苷中α-葡萄糖苷酶抑制剂活性成分及分子对接研究,评价其体内降糖活性。方法以阿卡波糖为阳性对照,半数抑制浓度(IC50)为α-葡萄糖苷酶抑制活性评价指标,建立体外α-葡萄糖苷酶抑制模型。运用UHPLC-Q-Exactive Orbitrap MS技术筛选并鉴定白扁豆总皂苷中的α-葡萄糖苷酶抑制剂活性成分,进一步对其中主要活性单体进行活性验证和酵母源α-葡萄糖苷酶和人源性α-葡萄糖苷酶的同源建模及分子对接,采用糖尿病小鼠模型评价其降糖活性。结果从白扁豆总皂苷中筛选并鉴定出了竹节参皂苷IVa等15个具有潜在α-葡萄糖苷酶抑制活性的化合物,同时对其中主要活性单体竹节参皂苷IVa进行活性验证,发现其有较强α-葡萄糖苷酶抑制活性,α-葡萄糖苷酶的IC50为(565.2±1.026)μg/m L低于阳性对照的阿卡波糖的IC50为(706.6±1.058)μg/m L,竹节参皂苷IVa与酵母源α-葡萄糖苷酶和人源性α-葡萄糖苷酶分子对接结合能分别为–6.1和–7.7 kcal/mol,与两者都具有较强的结合活性,竹节参皂苷IVa显著降低丙氨酸氨基转氨酶(ALT)、天门冬氨酸氨基转氨酶(AST)、尿素(UREA)、肌肝(CREA)和胆固醇(CHO)水平(P<0.05)。此外,对糖尿病小鼠损伤的肝脏和胰腺细胞有一定的修复作用。结论研究将为从白扁豆总皂苷中筛选α-葡萄糖苷酶抑制剂及结构改造提供依据。

关 键 词：Baibiandou(Lablab Semen Album) Total saponins UHPLC-Q-Exactive Orbitrap MS Α-GLUCOSIDASE Molecular docking Type 2 diabetic mice Chikusetsusaponin IVa 

分 类 号：R284[医药卫生—中药学]