Preparation and Characterization of  var. and  var. Extract and Analysis of Their Hemolysis

DING Li-shuai; ZHAO Meng; LI Yan-min; CHEN Liang-mian; WANG Zhu-ju; GAO Hui-min; WANG Zhi-min

doi:10.13422/j.cnki.syfjx.2017210007

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Preparation and Characterization of var. and var. Extract and Analysis of Their Hemolysis

更新时间：2024-01-04

- Preparation and Characterization of var. and var. Extract and Analysis of Their Hemolysis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 21, Pages: 7-12(2017)
- 作者机构：
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- DOI：10.13422/j.cnki.syfjx.2017210007
  CLC： R284
- Published：2017
- 稿件说明：
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DING Li-shuai, ZHAO Meng, LI Yan-min, et al. Preparation and Characterization of var. and var. Extract and Analysis of Their Hemolysis [J]. Chinese journal of experimental traditional medical formulae, 2017, 23(21): 7-12.
DOI：

DING Li-shuai, ZHAO Meng, LI Yan-min, et al. Preparation and Characterization of var. and var. Extract and Analysis of Their Hemolysis [J]. Chinese journal of experimental traditional medical formulae, 2017, 23(21): 7-12. DOI： 10.13422/j.cnki.syfjx.2017210007.

摘要

目的：比较七叶一枝花和滇重楼提取物体外溶血作用强弱，探讨不同结构类型重楼皂苷的溶血作用特点。方法：采用75%乙醇对七叶一枝花和滇重楼根茎进行提取，采用大孔吸附树脂对各类物质进行富集，采用快速制备色谱技术对目标组分群进行定向制备，基于对照品对照和在线紫外吸收光谱对总取物、部位和组分进行化学信息表征，采用分光光度法测定各样品的溶血率。结果：根据质量浓度-溶血率曲线计算七叶一枝花和滇重楼总提物的50%溶血浓度分别为34.97，46.52 mg·L-1。在2.5~10 mg·L-1时，七叶一枝花部位中仅有70%乙醇洗脱部位（PPC-D）显示了较强溶血作用，滇重楼部位中70%乙醇和95%乙醇洗脱部位（PPY-D和PPY-E）均显示溶血作用。5.0，10 mg·L-1时PPC-D的溶血率分别为78.00%和95.68%，PPY-D（PPY-E）的溶血率分别为31.86%和63.29%（15.82%和55.25%）。在2.5~5.0 mg·L-1时，七叶一枝花的组分呋甾皂苷和薯蓣皂苷未显示溶血作用，偏诺皂苷组分在质量浓度为2.5，5.0 mg·L-1的溶血率分别为5.89%和90.33%；滇重楼的呋甾皂苷组分同样未显示溶血作用，偏诺皂苷（薯蓣皂苷）组分在质量浓度为2.5，5.0 mg·L-1的溶血率分别为0.51%和57.75%（4.29%和49.39%）。结论：七叶一枝花和滇重楼均具有溶血作用，其中七叶一枝花强于滇重楼；七叶一枝花溶血成分群主要为偏诺皂苷类，滇重楼溶血成分群为偏诺皂苷和薯蓣皂苷；两者呋甾皂苷均无溶血作用；总提物溶血作用强弱主要由其中所含偏诺皂苷和薯蓣皂苷类成分的绝对含量和相对比例来决定。

Abstract

Objective: To compare in vitro hemolytic effect of Paris polyphylla var. chinensis (PPC) and P. polyhoylla var. yunnanensis (PPY) extract and to explore hemolytic property of various subtypes of steroidal saponins from Paridis Rhizoma. Method: The rhizoma of PPC or PPY was extracted with 75% ethanol.The extract was subjected to chromatography on HPD100 macroporous resin to give fractions of PPC(Y)-A-E. The sub-fractions of PPC(Y)-D-F

PPC(Y)-D-P and PPC(Y)-D-D were obtained by flash chromatography.The chemical profiles of these extract

fractions and sub-fractions were analyzed by HPLC and the main chromatographic peaks were identified on the basis of the comparison of reference substances and on-line ultraviolet spectra.The hemolytic effect of these samples against the rabbit red blood cells were determined by UV-VIS. Result: The hemolytic curves of PPC and PPY were obtained by plotting the hemolytic effect against the corresponding concentrations and the concentrations up to 50% hemolytic rate were calculated as 34.97

46.52 mg·L-1

respectively.Among fractions of PPC-A-E

PPC-D displayed the potent hemolytic effect at concentrations from 2.5-10 mg·L-1.For fractions of PPY-A-E

both PPY-D and PPY-E showed hemolytic effect at the same tested concentrations.The hemolytic rates of PPC-D at the concentration of 5.0

10 mg·L-1 were 78.00% and 95.68%

whereas PPY-D and PPY-E had the hemolytic rates of 31.86% and 63.29% as well as 15.82% and 55.25%.For the sub-fractions from PPC

furostanol saponins and dioscin saponins had no hemolytic effect

while pennogenin saponins took on the hemolytic rates of 5.89% and 90.33% at the concentration of 2.5

5.0 mg·L-1.For the sub-fractions from PPY

furostanol saponins display similar result as that of furostanol saponins from PPC

whereas pennogenin and dioscin saponins were observed at concentration of 2.5

5.0 mg·L-1 with the hemolytic rates of 0.51% and 57.75% as well as 4.29% and 49.39%

respectively. Conclusion: It is found that both PPC and PPY have hemolytic effects

and PPC is stronger than PPY.The hemolytic effect of PPC is related to its main constituents(pennogenin saponins) and that of PPY is due to the conjunction of pennogenin and dioscin saponins.The furostanol saponins from both plants have no hemolytic effect.The hemolytic level of PPC or PPY is determined according to the absolute and relative amounts of pennogenin and dioscin saponins in the extract.

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