Hepatoprotective Chemical Constituents from Thylacetate Extract of  Radix

LEI Zhong; ZHU Xin-yu; YANG Yu-sha; ZHOU Wei; LIANG Yan; HAO Xiao-yan

doi:10.13422/j.cnki.syfjx.20180609

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Hepatoprotective Chemical Constituents from Thylacetate Extract of Radix

更新时间：2024-01-04

- Hepatoprotective Chemical Constituents from Thylacetate Extract of Radix
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 24, Issue 7, Pages: 56-63(2018)
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- DOI：10.13422/j.cnki.syfjx.20180609
  CLC： R284.1
- Published：2018
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LEI Zhong, ZHU Xin-yu, YANG Yu-sha, et al. Hepatoprotective Chemical Constituents from Thylacetate Extract of Radix[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(7): 56-63.
DOI：

LEI Zhong, ZHU Xin-yu, YANG Yu-sha, et al. Hepatoprotective Chemical Constituents from Thylacetate Extract of Radix[J]. Chinese journal of experimental traditional medical formulae, 2018, 24(7): 56-63. DOI： 10.13422/j.cnki.syfjx.20180609.

摘要

目的：系统开展血人参乙酸乙酯部位保肝活性化学成分的研究工作。方法：将血人参的乙醇提取浸膏，依次进行萃取得到乙酸乙酯、正丁醇部位。开展在小鼠体内的保肝活性筛选试验，设立正常组、四氯化碳模型组、联苯双酯组、乙酸乙酯灌胃组和皮下组、正丁醇灌胃组和皮下组，检测不同组小鼠血清中谷丙转氨酶（ALT）活性，评价待测药物保肝活性。针对保肝活性部位，利用硅胶柱，Sephadex LH-20，反相C18色谱柱等手段进行活性单体化合物的分离，并进行结构鉴定。在四氯化碳诱导人肝HL-7702细胞株损伤模型下，对分离出来的化合物进行体外保肝活性筛选。结果：皮下给予血人参乙酸乙酯提取物对四氯化碳造成的小鼠肝损伤具有显著保护作用（P<0.05）。从血人参乙酸乙酯部分分离得到了11个化合物，经结构鉴定，确定为豆甾醇（化合物1），L-高丽槐素（化合物2），medicarpin（化合物3），豆甾4-烯-3-酮（化合物4），豆甾烷-3-酮（化合物5），3-hydroxy-8，9-dimethoxypterocarpan（化合物6），表儿茶素（化合物7），2α，3α-epoxyflavan-5，7，3'，4-tetraol-（4β→8）-flavan-5（化合物8），2α，3α-epoxy-5，7，3'，4'-tetrahydroxyflavan-（4β→8）-epicatechin（化合物9），表儿茶素（化合物10）和schizandriside（化合物11）。其中化合物2首次从血人参中分离得到，化合物3，化合物6首次从木蓝属植物中分离得到。化合物8，化合物9和化合物11对四氯化碳诱导的人肝HL-7702细胞损伤有一定程度的保护作用（P<0.05），HL-7702细胞存活率明显升高，提示化合物8，化合物9和化合物11在体外对肝细胞损伤有保护作用。结论：该研究确定血人参乙酸乙酯部位是该药材保肝活性的主要部位，而从中分离出的化合物8，化合物9和化合物11是血人参保肝活性的有效化合物。

Abstract

Objective: To systematically screen the hepatoprotective compounds from ethyl acetate extract of the roots of Indigofera stachyoides. Method: The roots of I. stachyoides were extracted by ethanol

and then its extract was further divided into ethyl acetate fraction and n-butanol fraction. The hepatoprotective activity screening was performed in mice in the normal control group

carbon tetrachloride (CCl4) model group

bifendate (DDB) group

intragastric administration group and subcutaneous group of ethyl acetate fraction

intragastric administration group and subcutaneous group of the n-butanol fraction. Glutamic pyruvic transaminase (ALT) level was detected in miceto evaluate the hepatoprotective activity of different samples. Then effective compounds of the hepatoprotective sample had been isolated and their structures were identified by silica gel column chromatography

Sephadex LH-20

RP-18 column chromatography and other means. Screening of hepatoprotective activity of isolated chemical compounds was performed in vitro in CCl4-induced human HL-7702 cell damage model. Result: Subcutaneous group of ethyl acetate fraction showed significant hepatoprotective effect on mice liver injury model (P<0.05). Total 11 compounds were isolated from this ethyl acetate fraction of I. stachyoides

which had been identified as stigmasterol (compound 1 )

L-maackiain (compound 2 )

medicarpin (compound 3 )

stigmast-4-en-3-one (compound 4 )

stigmastan-3-one (compound 5 )

3-hydroxy-8

9-dimethoxypterocarpan(compound 6 )

(+)-epicatechin(compound 7 )

2α

3α-epoxyflavan-5

4'-tetraol-(4β→8)-flavan-5″

7″

-4'"-triol (compound 8 )

2α

3α-epoxy-5

4'-tetrahydroxyflavan-(4β→8)-epicatechin (compound 9 )

(-)-epicatechin (compound 10 ) and schizandriside (compound 11 ). Compound 2 was isolated from this plant for the first time. Compounds 3 and 6 were isolated from Indigofera for the first time. Compounds 8

9 and 11 had a certain degree of protective effect on CCl4-induced human HL-7702 cell damage model (P<0.05). The survival rate of HL-7702 was significantly increased

suggesting that compound 8

compound 9 and compound 11 had protective effects on hepatocyte injury in vitro. Conclusion: In this study

ethyl acetate fraction of I. stachyoides was the main effective part of the hepatoprotective effect

and compound 8

9 and 11 from this fraction were the main effective compounds.

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