冻干与生晒西洋参饮片差异皂苷含量及促斑马鱼血管生成活性比较

邵娟; 徐梦丹; 张雪晶; 郭云龙; 蔡广知; 贡济宇

doi:10.13422/j.cnki.syfjx.20240768

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冻干与生晒西洋参饮片差异皂苷含量及促斑马鱼血管生成活性比较

药学基础 | 更新时间：2024-07-31

- 冻干与生晒西洋参饮片差异皂苷含量及促斑马鱼血管生成活性比较
  增强出版
- Comparison of Differential Saponin Contents and Pro-angiogenic Activities on Zebrafish Between Freeze-dried and Sun-dried Panacis Quinquefolii Radix Decoction Pieces
- 中国实验方剂学杂志 2024年30卷第17期页码：185-192
- 作者机构：
  
  1.长春中医药大学药学院，长春 130117
  2.长春中医药大学吉林省人参科学研究院，长春 130117
- 作者简介：
  
  邵娟，在读硕士，从事中药品质鉴定、质量标准及开发利用研究，E-mail：shaojuan07@163.com
  蔡广知，博士，副教授，从事中药品质鉴定、质量标准及开发利用研究，Tel：0431-86172193，E-mail：caigz@ccucm.edu.cn
- 基金信息：
  
  吉林省科技发展计划项目(20240305014YY)
- DOI：10.13422/j.cnki.syfjx.20240768
  中图分类号： R22;R825.4;R28;R943.1
- 纸质出版日期：2024-09-05，
  
  网络出版日期：2024-03-12，
  
  收稿日期：2024-01-08，
- 稿件说明：
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邵娟,徐梦丹,张雪晶等.冻干与生晒西洋参饮片差异皂苷含量及促斑马鱼血管生成活性比较[J].中国实验方剂学杂志,2024,30(17):185-192.

SHAO Juan,XU Mengdan,ZHANG Xuejing,et al.Comparison of Differential Saponin Contents and Pro-angiogenic Activities on Zebrafish Between Freeze-dried and Sun-dried Panacis Quinquefolii Radix Decoction Pieces[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(17):185-192.
邵娟,徐梦丹,张雪晶等.冻干与生晒西洋参饮片差异皂苷含量及促斑马鱼血管生成活性比较[J].中国实验方剂学杂志,2024,30(17):185-192. DOI： 10.13422/j.cnki.syfjx.20240768.

SHAO Juan,XU Mengdan,ZHANG Xuejing,et al.Comparison of Differential Saponin Contents and Pro-angiogenic Activities on Zebrafish Between Freeze-dried and Sun-dried Panacis Quinquefolii Radix Decoction Pieces[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(17):185-192. DOI： 10.13422/j.cnki.syfjx.20240768.

摘要

目的

分析和测定冻干与生晒西洋参饮片中的差异成分，并比较二者促血管生成活性的差异。

方法

基于超高效液相色谱法（UPLC）建立冻干和生晒西洋参饮片的指纹图谱，结合主成分分析（PCA）、正交偏最小二乘法-判别分析（OPLS-DA）等化学计量学方法确定2种饮片的差异皂苷成分，采用UPLC测定其

中6种代表性成分在冻干与生晒西洋参饮片中的含量；选取受精24 h的转基因斑马鱼品系Tg（fli1a∶EGFP）胚胎，分别使用不同剂量的冻干与生晒西洋参70%甲醇提取物（10、30 mg·L

-1

）干预正常斑马鱼和血管内皮生长因子受体酪氨酸激酶抑制剂Ⅱ（PTK787）诱导的节间血管（ISV）损伤斑马鱼模型，观察斑马鱼肠下静脉（SIV）和ISV发育情况，测定SIV直径、平均交叉数和平均发芽数，计算ISV血管指数，比较2种饮片的促血管生成活性。

结果

冻干与生晒西洋参饮片指纹图谱相似度均

0.950，分别确定了17个共有峰，指认了其中6个共有峰分别为峰6（人参皂苷Rg

）、峰7（人参皂苷Re）、峰8（人参皂苷Rb

）、峰11（人参皂苷Rc）、峰13（人参皂苷Rb

）、峰16（人参皂苷Rd）；PCA及OPLS-DA筛选出11种差异性皂苷成分，表明2种饮片的成分含量存在一定差异；含量测定结果显示，冻干西洋参饮片中人参皂苷Rg

、Re、Rb

、Rb

含量高于生晒西洋参饮片，而人参皂苷Rc、Rd含量低于生晒西洋参饮片（

0.05，

0.01）。在对正常斑马鱼胚胎促血管生成作用研究中，与空白组比较，冻干及生晒西洋参高剂量组SIV血管直径、平均发芽率及平均交叉率均显著升高（

0.01），且冻干西洋参组血管直径、平均交叉数及平均发芽数均高于生晒西洋参组（

0.05）。对ISV损伤的斑马鱼胚胎促血管生成作用研究显示，与空白组比较，模型组斑马鱼ISV发育受到抑制；与模型组比较，冻干及生晒西洋参各剂量组均可促进ISV的生长与萌发，相同剂量下，冻干西洋参组的正常血管数目明显高于生晒西洋参组（

0.05）。

结论

冻干处理西洋参饮片可有效避免其中人参皂苷类成分的流失与二次转化，其促斑马鱼血管生成活性优于生晒西洋参饮片，可为优质西洋参饮片的生产与开发提供参考。

Abstract

Objective

To analyze and determine the differential components of freeze-dried and sun-dried Panacis Quinquefolii Radix（PQR）， and to compare the differences in their pro-angiogenic activities.

Method

Fingerprints of freeze-dried and sun-dried PQR were established based on ultra performance liquid chromatography（UPLC）， and chemometrics methods such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were combined to determine the differential saponin composition of the two decoction pieces， and six representative saponins were selected and their contents in freeze-dried and sun-dried PQR were determined by UPLC. Transgenic zebrafish line Tg（fli1a∶EGFP） embryos fertilized for 24 h were selected， and different doses of 70% methanol extracts of freeze-dried and sun-dried PQR（10， 30 mg·L

-1

） were used to intervene in normal

zebrafish and in a zebrafish model of intersegmental vascular（ISV） injury induced by vascular endothelial growth factor（VEGF） receptor tyrosine kinase inhibitor Ⅱ（PTK787）， then the development of subintestinal vein（SIV） and ISV of zebrafish was observed， SIV diameter， mean number of crossings and mean number of germinations were determined， and the ISV vascular index was calculated， in order to compare the pro-angiogenic activities of the two decoction pieces.

Result

The similarity of the fingerprints of freeze-dried and sun-dried PQR decoction pieces was

0.950， and 17 common peaks were identified， of which 6 common peaks were designated as peak 6（ginsenoside Rg

）， peak 7（ginsenoside Re）， peak 8（ginsenoside Rb

）， peak 11（ginsenoside Rc）， peak 13（ginsenoside Rb

）， and peak 16（ginsenoside Rd）， respectively. A total of 11 differential saponin components were screened by PCA and OPLS-DA， indicating that there were some differences in the contents of the components in the two decoction pieces. The results of determination showed that the contents of ginsenosides Rg

， Re， Rb

and Rb

in freeze-dried PQR were higher than those in sun-dried PQR， while the contents of ginsenosides Rc and Rd were lower than those in sun-dried PQR（

0.05，

0.01）. In the study of the pro-angiogenic effect on normal zebrafish embryos， compared with the blank group， and the SIV vessel diameter， mean germination rate and mean crossover rate were significantly higher in the high-dose groups of freeze-dried and sun-dried PQR（

0.01）， and the vessel diameter， mean numbers of crossings and germinations in the freeze-dried PQR group were higher than those of the sun-dried PQR group（

0.05）. In the study of the pro-angiogenic effect on zebrafish embryos with ISV injury， the development of ISV in the model group was significantly inhibited when compared with the blank group

， compared with the model group， different dose groups of freeze-dried and sun-dried PQR could promote the growth and sprouting of ISV， and the number of normal blood vessels in the freeze-dried PQR group was significantly higher than that in the sun-dried PQR group at the same dosage（

0.05）.

Conclusion

Freeze-drying can effectively avoid the loss and secondary transformation of ginsenosides in PQR， and its angiogenic activity is better than that of sun-dried PQR， which can provide a reference for the production and development of high-quality PQR decoction pieces.

关键词

西洋参饮片冻干生晒指纹图谱含量测定化学计量法促血管生成

Keywords

Panacis Quinquefolii Radix decoction piecesfreeze-dryingsun-dryingfingerprintdeterminationstoichiometrypromoting angiogenesis

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