Selective Inhibitory Effect of Typhae Pollen and Paeoniae Radix Rubra on Platelet Aggregation Based on Activity Screening and Target Network Prediction

WANG Xiao-yi; TIAN Xiao-xuan; ZHANG Yan; WU Hong-hua; WANG Yue-fei; ZHU Yan

doi:10.13422/j.cnki.syfjx.2017010120

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Selective Inhibitory Effect of Typhae Pollen and Paeoniae Radix Rubra on Platelet Aggregation Based on Activity Screening and Target Network Prediction

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- Selective Inhibitory Effect of Typhae Pollen and Paeoniae Radix Rubra on Platelet Aggregation Based on Activity Screening and Target Network Prediction
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 23, Issue 1, Pages: 120-126(2017)
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- DOI：10.13422/j.cnki.syfjx.2017010120
  CLC： R285
- Published：2017
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WANG Xiao-yi, TIAN Xiao-xuan, ZHANG Yan, et al. Selective Inhibitory Effect of Typhae Pollen and Paeoniae Radix Rubra on Platelet Aggregation Based on Activity Screening and Target Network Prediction[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(1): 120-126.
DOI：

WANG Xiao-yi, TIAN Xiao-xuan, ZHANG Yan, et al. Selective Inhibitory Effect of Typhae Pollen and Paeoniae Radix Rubra on Platelet Aggregation Based on Activity Screening and Target Network Prediction[J]. Chinese journal of experimental traditional medical formulae, 2017, 23(1): 120-126. DOI： 10.13422/j.cnki.syfjx.2017010120.

摘要

目的：探讨常用活血化瘀中药抗血小板活性的激动剂选择性及可能的作用通路。方法：通过体外高通量血小板聚集实验系统评价24种常用活血化瘀中药醇提取物抗二磷酸腺苷（ADP）和凝血酶诱导的血小板聚集活性；通过Ingenuity Pathway Analysis（IPA）预测代表性药物蒲黄与赤芍已知单体成分的作用靶点，并加以验证。结果：蒲黄抗ADP，凝血酶诱导血小板聚集的半数抑制浓度（IC50）分别为55.27，300.60 mg·L-1；赤芍抗ADP，凝血酶诱导血小板聚集的IC50分别为336.20，101.60 mg·L-1。IPA预测提示，蒲黄与赤芍有可能通过调节Ca2+，环磷酸腺苷（cAMP）和一氧化氮（NO）水平抑制血小板聚集。实验发现，蒲黄可明显抑制ADP对血小板内Ca2+的升高（P < 0.05，P < 0.01）；赤芍可显著提高凝血酶导致的血小板内NO浓度降低（P<0.01）；蒲黄、赤芍均不能逆转ADP，凝血酶引起的血小板内cAMP浓度降低。结论：蒲黄和赤芍醇提物分别具有选择性抗ADP和凝血酶诱导的大鼠血小板聚集活性。与IPA预测相符，蒲黄可降低血小板内钙离子水平，赤芍可提高血小板内NO浓度，可能分别激活嘌呤能P2Y1受体下游的Gq亚基/磷脂酶C（PLC）/三磷酸肌醇（IP3）和磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（Akt）/内皮型一氧化氮合酶（eNOS）通路抑制血小板聚集。

Abstract

Objective: To investigate the selective inhibitory effect of the Chinese medicines with functions of promoting blood circulation and removing blood stasis on platelet aggregation

and explore the underlying mechanism. Method: Ethanol extracts of 24 kinds of common promoting blood circulation and removing blood stasis Chinese medicines were evaluated for adenosine diphosphate (ADP) and thrombin-induced platelet aggregation activity by using in vitro high throughput platelet aggregation assay. Ingenuity Pathway Analysis (IPA) was used to predict and verify the potential targets of the known monomer components in representative herbs Typhae Pollen and Paeoniae Radix Rubra. Result: The IC50 of Typhae Pollen on platelet aggregation induced by ADP and thrombin was 55.27

300.60 mg·L-1 respectively

while the IC50 of Paeoniae Radix Rubra was 336.20

101.60 mg·L-1 respectively. IPA prediction showed that Typhae Pollen and Paeoniae Radix Rubra inhibited platelet aggregation probably by regulating the calcium mobilization (Ca2+)

cyclic adenosine monophosphate (cAMP) and nitric oxide (NO) levels. The results showed that Typhae Pollen could significantly inhibit the level increase of Ca2+ in platelet stimulated by ADP (P < 0.05

P < 0.01). Paeoniae Radix Rubra could significantly increase the level decrease of NO in platelet induced by thrombin (P<0.01). Both of Typhae Pollen and Paeoniae Radix Rubra could not reverse the level decrease of cAMP induced by ADP or thrombin. Conclusion: The ethanol extracts of Typhae Pollen electively inhibited the platelet aggregation induced by ADP

while the ethanol extracts of Paeoniae Radix Rubra selectively inhibited the platelet aggregation induced by thrombin. As the prediction by IPA

Typhae Pollen could decrease the level of Ca2+ and Paeoniae Radix Rubra could increase the level of NO

and they could inhibit the platelet aggregation probably by activating Gq/phospholipase C(PLC)/inositol triphosphate(IP3) of P2Y1

and phosphatidylinositide-3-kinases (PI3K)/Akt/endothelial nitric oxide synthase (eNOS) pathways respectively.

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Related Institution

The First Affiliated Hospital of Hunan University of Chinese Medicine

Tianjin University of Traditional Chinese Medicine（TCM）

National Resource Center for Chinese Materia Medica，State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs， China Academy of Chinese Medical Sciences

Institute of TCM Health Industry，China Academy of Chinese Medical Sciences

College of Pharmacy，Hebei University of Chinese Medicine

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