9种通络祛风中药提取物对黄嘌呤氧化酶的体外抑制活性研究
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篇名: 9种通络祛风中药提取物对黄嘌呤氧化酶的体外抑制活性研究
TITLE: Study on in vitro Inhibitory Activity of the Extracts from 9 Kinds of TCM for Dredging Collaterals and Dispelling Wind on Xanthine Oxidase
摘要: 目的:研究9种通络祛风中药的提取物对黄嘌呤氧化酶(XO)的体外抑制活性,以筛选活性突出的中药提取物。方法:以黄嘌呤为底物、XO为反应酶,以别嘌醇为阳性对照,以三角风、石胡椒、山莓、金雀花根、紫藤根、牛迭肚根、梓木皮、凌霄花根、爬岩香等药材的水提物和甲醇提取物(以下简称“醇提物”)以及活性提取物的石油醚、二氯甲烷、乙酸乙酯、正丁醇、水等萃取部位为对象,采用紫外分光光度法检测各样品对XO的抑制率,采用Graphpadprism6.0软件计算半数抑制浓度(IC50)以筛选活性提取物/部位;采用双倒数法判断药材的酶抑制作用的类型。结果:9种中药共18个提取物中,500μg/mL各中药提取物(除石胡椒醇提物外),250μg/mL三角风、石胡椒、山莓、爬岩香的水提物和醇提物,250μg/mL梓木皮水提物,250μg/mL金雀花根、牛迭肚根、凌霄花根醇提物,125μg/mL金雀花根、牛迭肚根醇提物以及62.5μg/mL金雀花根醇提物对XO的抑制率均超过了50%;其中,金雀花根醇提物的IC50值为43.43μg/mL,低于其他药材的提取物,为活性提取物。金雀花根醇提物石油醚、二氯甲烷、乙酸乙酯、正丁醇、水等萃取部位的IC50值分别为>200、193.35、7.67、14.80、>200μg/mL,其中乙酸乙酯提取部位的IC50值与阳性对照(5.11μg/mL)接近。金雀花根醇提物对XO的抑制类型为竞争-非竞争性抑制,不同于阳性对照的竞争性抑制。结论:金雀花根、牛迭肚根、石胡椒、山莓、爬岩香、三角风等醇提物及三角风、石胡椒、梓木皮等水提物对XO均具有一定的体外抑制活性,以金雀花根醇提物最强,且该提取物的乙酸乙酯萃取部位的抑制活性与别嘌醇相当,但抑制类型有所差异。
ABSTRACT: OBJECTIVE:To study in vi tro inhibitory act ivities of 9 kinds of TCM for dredging collaterals and dispelling wind on xanthine oxidase (XO),and to screen TCM with outstanding activity. METHODS :Using xanthine as substrate and xanthinase as reaction enzyme ,allopurinol as positive control ,with water extract and methanol extract (hereinafter referred to as “ethanol extract”)from the stem and leaves of Hedera nepalensis ,the whole plant of Piper wallichii ,the fruits of Rubus corchorifolius ,the root of Caragana sinica ,the root of Wisteria sinensis ,the root of Rubus crataegifolius ,the bark of Catalpa ovata ,the root of Campsis grandiflora ,the stem of P. hancei (hereinafter referred to by plant name )and petroleum ether ,dichloromethane,ethyl acetate,n-butanol and water fraction of active extract as the objects ,inhibition rate of each sample to XO was detected by spectrophotometry;IC50values were calculated with Graphpad prism 6.0 software to screen active extract/fraction. Double reciprocal method was used to determine the type of enzyme inhibition. RESULTS :Among 9 kinds of TCM and 18 kinds of the extracts ,the inhibitory rates to XO of 500 μg/mL extracts from each TCM(except for ethanol extract of P. wallichii ),250 μg/mL water extract and ethanol extract of H. nepalensis ,P. wallichii ,R. corchorifolius and P. hancei ,250 μg/mL water extract of C. ovata ,250 μ g/mL ethanol extract of C. sinica ,R. crataegifolius and C. grandiflora ,125 μ g/mL ethanol extract of C. sinica and R. crataegifolius,62.5 μg/mL ethanol extract of C. sinica were more than 50%. The IC 50 value of the ethanol extract from C. sinica was 43.43 μg/mL,which was lower than the extracts of other TCM ,and which was the active extract. The IC 50 values of petroleum ether,dichloromethane,ethyl acetate ,n-butanol and water fracti on of ethanol extract from C. sinica were >200,193.35,7.67, 14.80 and >200 μg/mL,respectively. The IC 50 value of ethyl XO was competitive-noncompetitive inhibition , which was different from competi tive inhibition of positive control. CONCLUSIONS:The ethanol extracts of C. sinica ,R. crataegifolius ,P. wallichii ,R. corchorifolius ,P. hancei ,H. nepalensis and the water extracts of H. nepalensis ,P. wallichii ,C. ovata show certain inhibitory activity in vitro to XO ,especially ethanol extract of C. sinica . The ethyl acetate fraction of the ethanol extract of C. sinica has similar inhibitory activity to allopurinol but their inhibition types are different.
期刊: 2020年第31卷第06期
作者: 李芮,马良会,王栋,张春雷,周孟,廖尚高
AUTHORS: LI Rui,MA Lianghui ,WANG Dong,ZHANG Chunlei ,ZHOU Meng,LIAO Shanggao
关键字: 黄嘌呤氧化酶抑制活性;痛风;中药;提取物;金雀花根;体外
KEYWORDS: Xanthine oxidase inhibitory activity ;Gout;TCM;Extract;Root of Caragana sinica ;in vitr o
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