超滤亲和-液质联用和分子对接技术筛选毛菊苣根中α-葡萄糖苷酶抑制剂
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摘要
目的采用超滤亲和-液质联用(Ultra-filtration affinity-liquid chromatography-mass spectrometry,UF-LC-MS)和分子对接技术筛选毛菊苣Cichorium glandulosum根中α-葡萄糖苷酶抑制剂。方法通过超滤亲和-液质联用技术筛选并鉴定毛菊苣根中的α-葡萄糖苷酶抑制剂,发现有4个组分对α-葡萄糖苷酶的活性具有抑制作用,进一步采用分子对接技术对这4个化合物进行验证。结果发现2个化合物对α-葡萄糖苷酶具有较高的抑制作用,其中黄芩苷、山莴苣苦素在0.1～2.0mg/mL呈良好的量效关系。结论为利用UF-LC-MS和分子对接技术筛选天然产物中α-葡萄糖苷酶抑制剂提供了参考,也为后续研究毛菊苣降糖功效提供了基础。
Objective To screen α-glucosidase inhibitors from root extract of Cichorium glundulosum(CGR) by UF-LC-MS and molecular docking.Methods The inhibitory activity of CGR extract was evaluated by establishing enzyme inhibitor model,Meanwhile,four active compounds from CGR extract were screened by UF-LC-MS and identified as α-glucosidase inhibitors,which were verified by using Autodock Software.Results The results showed that two compounds that combined with α-glucosidase well were screened out,including baicalin and lactupicrin,which showed significant α-glucosidase inhibitory activity in the concentration range of 0.1—2 mg/mL in a dose-dependent manner.Conclusion Our work provide theoretical basis for screening natural and high affinity enzyme inhibitors from CGR by using UF-LC-MS and molecular docking and further study of the hypoglycemic effect of C.glundulosum.
Objective To screen α-glucosidase inhibitors from root extract of Cichorium glundulosum(CGR) by UF-LC-MS and molecular docking.Methods The inhibitory activity of CGR extract was evaluated by establishing enzyme inhibitor model,Meanwhile,four active compounds from CGR extract were screened by UF-LC-MS and identified as α-glucosidase inhibitors,which were verified by using Autodock Software.Results The results showed that two compounds that combined with α-glucosidase well were screened out,including baicalin and lactupicrin,which showed significant α-glucosidase inhibitory activity in the concentration range of 0.1—2 mg/mL in a dose-dependent manner.Conclusion Our work provide theoretical basis for screening natural and high affinity enzyme inhibitors from CGR by using UF-LC-MS and molecular docking and further study of the hypoglycemic effect of C.glundulosum.
引文
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[4]张尧,信学雷,阿吉艾克拜尔·艾萨,等.维药毛菊苣降糖有效部位的纯化工艺[J].中成药,2012,34(12):2322-2325.
[5]Tong J,Ma B,Ge L,et al.Dicaffeoylquinic acid-enriched fraction of Cichorium glandulosum seeds attenuates experimental type 1 diabetes via multipathway protection[J].J Agric Food Chem,2015,63(50):10791-10799.
[6]姚红锐,龙子江,赵刚,等.毛菊苣有效部位降脂作用及初步机制探讨[J].安徽医药,2015,19(10):1857-1860.
[7]Upur H,Amat N,Bla?Ekovic’B,et al.Protective effect of Cichorium glandulosum root extract on carbon tetrachloride-induced and galactosamine-induced hepatotoxicity in mice[J].Food Chem Toxicol,2009,47(8):2022-2030.
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[9]Chai T T,Kwek M T,Ong H C,et al.Water fraction of edible medicinal fern Stenochlaena palustris is a potentα-glucosidase inhibitor with concurrent antioxidant activity[J].Food Chem,2015,186(2/3):26-31.
[10]Chen G,Guo M.Rapid screening forα-glucosidase inhibitors from Gymnema sylvestre by affinity ultrafiltration-HPLC-MS[J].Frontiers Pharmacol,2017,30(S1):95-99.
[11]Yang J B,Tian J Y,Dai Z,et al.a-Glucosidase inhibitors extracted from the roots of Polygonum multiflorum Thunb[J].Fitoterapia,2016,117(7):65-67.
[12]Nguyen V B,Nguyen Q V,Nguyen A D,et al.Screening and evaluation ofα-glucosidase inhibitors from indigenous medicinal plants in Dak Lak Province,Vietnam[J].Res Chem Intermediates,2016,43(6):3599-3602.
[13]Trinh B T,Staerk D,Jager A K.Screening for potential alpha-glucosidase and alpha-amylase inhibitory constituents from selected Vietnamese plants used to treat type 2 diabetes[J].J Ethnopharmacol,2016,186(32):189-195.
[14]Yan H,Qiu C,Zhong K,et al.Inhibition ofα-glucosidase activity by 3,5-dicaffeoylquinic acid from Lonicera japonica thunb flower buds[J].Modern Food Sci Tech,2015,31(7):44-49.
[15]范莉,王业玲,唐丽.天然来源α-葡萄糖苷酶抑制剂筛选方法的研究进展[J].天然产物研究与开发,2016,46(2):313-321.
[16]Kim J H,Ahn J H,Kim S K,et al.Combined use of basal insulin analog and acarbose reduces postprandial glucose in patients with uncontrolled type 2 diabetes[J].JDiabetes Investigation,2015,6(2):219-226.
[17]Yang D,Zhao J,Liu S,et al.The screening of potentialα-glucosidase inhibitors from the Polygonum multiflorum extract using ultrafiltration combined with liquid chromatography-tandem mass spectrometry[J].Anal Methods,2014,6(10):3353-3359.
[18]Zhang A,Ye F,Lu J,et al.Screeningα-glucosidase inhibitor from natural products by capillary electrophoresis with immobilised enzyme onto polymer monolith modified by gold nanoparticles[J].Food Chem,2013,141(3):1854-1859.
[19]邬思琪,杨华,李萍.亲和超滤结合液质技术在中药有效成分发现中的应用[J].药学学报,2016,51(7):1060-1067.
[20]张加余,屠鹏飞.天然产物液相色谱-质谱-数据库(LC-MS-DS)的建立与应用[J].药学学报,2012,45(9):1187-1192.
[21]Breemen R B V,Huang C R,Nikolic D,et al.Pulsed ultrafiltration mass spectrometry:A new method for screening combinatorial libraries[J].Anal Chem,1997,69(11):2159-2164.
[22]Wang J,Liu S,Ma B,et al.Rapid screening and detection of XOD inhibitors from S.tamariscina by ultrafiltration LC-PDA-ESI-MS combined with HPCCC[J].Anal Bioanalyt Chem,2014,406(28):7379-7387.
[23]Fang Y X,Song H P,Liang J X,et al.Rapid screening of pancreatic lipase inhibitors from Monascus-fermented rice by ultrafiltration liquid chromatography-mass spectrometry[J].Anal Methods,2017,23(9):3422-3429.
[24]Ding L,Liu J L,Hassan W,et al.Lipid modulatory activities of Cichorium glandulosum Boiss et Huet are mediated by multiple components within hepatocytes[J].Sci Reports,2014,4(17):4715-4717.
[25]杨飞,罗玉琴,薛桂蓬,等.维药毛菊苣根-种子不同比例配伍的90%乙醇提取物对小鼠急性肝损伤的保护作用及其机制[J].中国医院药学杂志,2015,35(15):1352-1356.
[26]唐英,刘春明,李森林,等.黄连中α-葡萄糖苷酶抑制剂的筛选分离及质谱分析[J].天然产物研究与开发,2016,28(7):1078-1083.
[27]Hui C,Ouyang K,Yan J,et al.Constituent analysis of the ethanol extracts of Chimonanthus nitens Oliv.leaves and their inhibitory effect onα-glucosidase activity[J].Inter JBiol Macromol,2017,98(9):829-836.
[28]Li H,Song F,Xing J,et al.Screening and structural characterization ofα-glucosidase inhibitors from hawthorn leaf flavonoids extract by ultrafiltration LC-DAD-MS n and SORI-CID FTICR MS[J].J Am Soc Mass Spectr,2009,20(8):1496-1503.
[29]Striegel L,Kang B,Pilkenton S J,et al.Apostolidis E.Effect of black tea and black tea pomace polyphenols on?±-glucosidase and?±-amylase inhibition,relevant to type2 diabetes prevention[J].Frontiers Nutr,2015,2(3):3-6.
[30]Deng S,Xia L,Xiao H.Screening ofα-glucosidase inhibitors from green tea extracts using immobilized enzymes affinity capture combined with UHPLC-QTOFMS analysis[J].Chem Comm,2014,50(20):2582-2584.
[31]Ji T,Li J,Su S L,et al.Identification and determination of the polyhydroxylated alkaloids compounds withα-glucosidase inhibitor activity in mulberry leaves of different origins[J].Molecules,2016,21(2):206-208.
[32]Luo L,Hao Y,Jia L,et al.Analysis of chlorogenic acid oxidation pathway in simulated enzymatic honeysuckle browning system by high performance liquid chromatography and mass spectrometry[J].Tropical JPharm Res,2016,15(2):405-407.
[33]Tóth G,Albertiá,Sólyomváry A,et al.Phenolic profiling of various olive bark-types and leaves:HPLC-ESI/MSstudy[J].Industr Crops Prods,2015,67(2):432-438.
[34]Gu Y X,Yu C H,Zhou Z L.Determination of baicalin in cosmetics and toothpaste by solid-phase extraction and high-performance liquid chromatography[J].Instrumentation Sci Technol,2016,44(6):593-602.
[35]Wang Y,Min Z,Ou Y,et al.Metabolic profile of esculin in rats by ultra high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry[J].J Chromatogr B,2016,1020(23):120-124.
[36]Yi T,Zhu L,Tang Y N,et al.An integrated strategy based on UPLC-DAD-QTOF-MS for metabolism and pharmacokinetic studies of herbal medicines:Tibetan“Snow Lotus”herb(Saussurea laniceps),a case study[J].J Ethnopharmacol,2014,153(3):701-713.
[37]辛文妤,宋俊科,何国荣,等.黄芩素和黄芩苷的药理作用及机制研究进展[J].中国新药杂志,2013,22(6):647-653.
[38]Sun M,Su M,Sun H.Spectroscopic investigation on the interaction characteristics and inhibitory activities between baicalin and acetylcholinesterase[J].Med Chem Res,2018,27(6):1589-1598.
[39]Xi Y,Wu M,Li H,et al.Baicalin attenuates high fat diet-induced obesity and liver dysfunction:Dose-response and potential role of CaMKKβ/AMPK/ACC pathway[J].Cell Physiol Biochem Inter J Exp Cell Physiol Biochem Pharmacol,2015,35(6):2349-2359.
[40]李小华,孙毅毅.秦皮甲素对高脂饮食诱导的大鼠脂肪肝的防治作用[J].中国新药与临床杂志,2010,29(9):679-683.
[2]潘兰,贾新岳,石明辉,等.维药毛菊苣化学成分研究[J].新疆医科大学学报,2015,45(9):1088-1091.
[3]信学雷,吴汉夔,吕俏莹,等.维吾尔药毛菊苣提取物降糖活性的研究[J].天然产物研究与开发,2012,24(2):234-238.
[4]张尧,信学雷,阿吉艾克拜尔·艾萨,等.维药毛菊苣降糖有效部位的纯化工艺[J].中成药,2012,34(12):2322-2325.
[5]Tong J,Ma B,Ge L,et al.Dicaffeoylquinic acid-enriched fraction of Cichorium glandulosum seeds attenuates experimental type 1 diabetes via multipathway protection[J].J Agric Food Chem,2015,63(50):10791-10799.
[6]姚红锐,龙子江,赵刚,等.毛菊苣有效部位降脂作用及初步机制探讨[J].安徽医药,2015,19(10):1857-1860.
[7]Upur H,Amat N,Bla?Ekovic’B,et al.Protective effect of Cichorium glandulosum root extract on carbon tetrachloride-induced and galactosamine-induced hepatotoxicity in mice[J].Food Chem Toxicol,2009,47(8):2022-2030.
[8]何茂群,高峰,鞠建刚.毛菊苣根乙酸乙酯提取部位的化学成分[J].辽宁中医药大学学报,2014,17(7):70-71.
[9]Chai T T,Kwek M T,Ong H C,et al.Water fraction of edible medicinal fern Stenochlaena palustris is a potentα-glucosidase inhibitor with concurrent antioxidant activity[J].Food Chem,2015,186(2/3):26-31.
[10]Chen G,Guo M.Rapid screening forα-glucosidase inhibitors from Gymnema sylvestre by affinity ultrafiltration-HPLC-MS[J].Frontiers Pharmacol,2017,30(S1):95-99.
[11]Yang J B,Tian J Y,Dai Z,et al.a-Glucosidase inhibitors extracted from the roots of Polygonum multiflorum Thunb[J].Fitoterapia,2016,117(7):65-67.
[12]Nguyen V B,Nguyen Q V,Nguyen A D,et al.Screening and evaluation ofα-glucosidase inhibitors from indigenous medicinal plants in Dak Lak Province,Vietnam[J].Res Chem Intermediates,2016,43(6):3599-3602.
[13]Trinh B T,Staerk D,Jager A K.Screening for potential alpha-glucosidase and alpha-amylase inhibitory constituents from selected Vietnamese plants used to treat type 2 diabetes[J].J Ethnopharmacol,2016,186(32):189-195.
[14]Yan H,Qiu C,Zhong K,et al.Inhibition ofα-glucosidase activity by 3,5-dicaffeoylquinic acid from Lonicera japonica thunb flower buds[J].Modern Food Sci Tech,2015,31(7):44-49.
[15]范莉,王业玲,唐丽.天然来源α-葡萄糖苷酶抑制剂筛选方法的研究进展[J].天然产物研究与开发,2016,46(2):313-321.
[16]Kim J H,Ahn J H,Kim S K,et al.Combined use of basal insulin analog and acarbose reduces postprandial glucose in patients with uncontrolled type 2 diabetes[J].JDiabetes Investigation,2015,6(2):219-226.
[17]Yang D,Zhao J,Liu S,et al.The screening of potentialα-glucosidase inhibitors from the Polygonum multiflorum extract using ultrafiltration combined with liquid chromatography-tandem mass spectrometry[J].Anal Methods,2014,6(10):3353-3359.
[18]Zhang A,Ye F,Lu J,et al.Screeningα-glucosidase inhibitor from natural products by capillary electrophoresis with immobilised enzyme onto polymer monolith modified by gold nanoparticles[J].Food Chem,2013,141(3):1854-1859.
[19]邬思琪,杨华,李萍.亲和超滤结合液质技术在中药有效成分发现中的应用[J].药学学报,2016,51(7):1060-1067.
[20]张加余,屠鹏飞.天然产物液相色谱-质谱-数据库(LC-MS-DS)的建立与应用[J].药学学报,2012,45(9):1187-1192.
[21]Breemen R B V,Huang C R,Nikolic D,et al.Pulsed ultrafiltration mass spectrometry:A new method for screening combinatorial libraries[J].Anal Chem,1997,69(11):2159-2164.
[22]Wang J,Liu S,Ma B,et al.Rapid screening and detection of XOD inhibitors from S.tamariscina by ultrafiltration LC-PDA-ESI-MS combined with HPCCC[J].Anal Bioanalyt Chem,2014,406(28):7379-7387.
[23]Fang Y X,Song H P,Liang J X,et al.Rapid screening of pancreatic lipase inhibitors from Monascus-fermented rice by ultrafiltration liquid chromatography-mass spectrometry[J].Anal Methods,2017,23(9):3422-3429.
[24]Ding L,Liu J L,Hassan W,et al.Lipid modulatory activities of Cichorium glandulosum Boiss et Huet are mediated by multiple components within hepatocytes[J].Sci Reports,2014,4(17):4715-4717.
[25]杨飞,罗玉琴,薛桂蓬,等.维药毛菊苣根-种子不同比例配伍的90%乙醇提取物对小鼠急性肝损伤的保护作用及其机制[J].中国医院药学杂志,2015,35(15):1352-1356.
[26]唐英,刘春明,李森林,等.黄连中α-葡萄糖苷酶抑制剂的筛选分离及质谱分析[J].天然产物研究与开发,2016,28(7):1078-1083.
[27]Hui C,Ouyang K,Yan J,et al.Constituent analysis of the ethanol extracts of Chimonanthus nitens Oliv.leaves and their inhibitory effect onα-glucosidase activity[J].Inter JBiol Macromol,2017,98(9):829-836.
[28]Li H,Song F,Xing J,et al.Screening and structural characterization ofα-glucosidase inhibitors from hawthorn leaf flavonoids extract by ultrafiltration LC-DAD-MS n and SORI-CID FTICR MS[J].J Am Soc Mass Spectr,2009,20(8):1496-1503.
[29]Striegel L,Kang B,Pilkenton S J,et al.Apostolidis E.Effect of black tea and black tea pomace polyphenols on?±-glucosidase and?±-amylase inhibition,relevant to type2 diabetes prevention[J].Frontiers Nutr,2015,2(3):3-6.
[30]Deng S,Xia L,Xiao H.Screening ofα-glucosidase inhibitors from green tea extracts using immobilized enzymes affinity capture combined with UHPLC-QTOFMS analysis[J].Chem Comm,2014,50(20):2582-2584.
[31]Ji T,Li J,Su S L,et al.Identification and determination of the polyhydroxylated alkaloids compounds withα-glucosidase inhibitor activity in mulberry leaves of different origins[J].Molecules,2016,21(2):206-208.
[32]Luo L,Hao Y,Jia L,et al.Analysis of chlorogenic acid oxidation pathway in simulated enzymatic honeysuckle browning system by high performance liquid chromatography and mass spectrometry[J].Tropical JPharm Res,2016,15(2):405-407.
[33]Tóth G,Albertiá,Sólyomváry A,et al.Phenolic profiling of various olive bark-types and leaves:HPLC-ESI/MSstudy[J].Industr Crops Prods,2015,67(2):432-438.
[34]Gu Y X,Yu C H,Zhou Z L.Determination of baicalin in cosmetics and toothpaste by solid-phase extraction and high-performance liquid chromatography[J].Instrumentation Sci Technol,2016,44(6):593-602.
[35]Wang Y,Min Z,Ou Y,et al.Metabolic profile of esculin in rats by ultra high performance liquid chromatography combined with Fourier transform ion cyclotron resonance mass spectrometry[J].J Chromatogr B,2016,1020(23):120-124.
[36]Yi T,Zhu L,Tang Y N,et al.An integrated strategy based on UPLC-DAD-QTOF-MS for metabolism and pharmacokinetic studies of herbal medicines:Tibetan“Snow Lotus”herb(Saussurea laniceps),a case study[J].J Ethnopharmacol,2014,153(3):701-713.
[37]辛文妤,宋俊科,何国荣,等.黄芩素和黄芩苷的药理作用及机制研究进展[J].中国新药杂志,2013,22(6):647-653.
[38]Sun M,Su M,Sun H.Spectroscopic investigation on the interaction characteristics and inhibitory activities between baicalin and acetylcholinesterase[J].Med Chem Res,2018,27(6):1589-1598.
[39]Xi Y,Wu M,Li H,et al.Baicalin attenuates high fat diet-induced obesity and liver dysfunction:Dose-response and potential role of CaMKKβ/AMPK/ACC pathway[J].Cell Physiol Biochem Inter J Exp Cell Physiol Biochem Pharmacol,2015,35(6):2349-2359.
[40]李小华,孙毅毅.秦皮甲素对高脂饮食诱导的大鼠脂肪肝的防治作用[J].中国新药与临床杂志,2010,29(9):679-683.