Comparing coagulation activity of Selaginella tamariscina before and after stir-frying process and determining the possible active constituents based on compositional variation

Pharm Biol. 2018 Dec;56(1):67-75. doi: 10.1080/13880209.2017.1421673.

Abstract

Context: Selaginella tamariscina (P. Beauv.) Spring (Selaginellaceae) (ST) has been widely used in China as a medicine for improving blood circulation. However, its processed product, S. tamariscina carbonisatus (STC), possesses opposite haemostatic activity.

Objective: To comprehensively evaluate the activity of ST and STC on physiological coagulation system of rats, and seek potential active substances accounting for the activity transformation of ST during processing.

Materials and methods: The 75% methanol extracts of the whole grass (fine powder) of ST and STC were prepared, respectively. Male Sprague-Dawley rats were randomly divided into five groups: control group, model group, model + ST group, model + STC group and positive control group (model + Yunnanbaiyao). The duration of intragastric administration was 72 h at 12 h intervals. Haemorheology parameters were measured using an LB-2 A cone-plate viscometer and the existed classic methods, respectively. SC40 semi-automatic coagulation analyzer was employed to determine coagulation indices. Meanwhile, HPLC and LC-MS were applied for chemical analyses of ST and STC extracts.

Results: STC shortened tail-bleeding time, increased whole blood viscosity (WBV) and plasma viscosity (PV), decreased erythrocyte sedimentation rate blood (ESR), reduced activated partial thromboplastin time (APTT) and increased the fibrinogen (FIB) content in the plasma of bleeding model rats. Although ST could shorten APTT and TT, the FIB content was significantly decreased by ST. Dihydrocaffeic acid with increased content in STC vs. ST showed haemostatic activity for promoting the platelet aggregation induced by collagen and trap-6, and reducing APTT and PT significantly with a concentration of 171.7 μM in vitro. Amentoflavone with reduced content in STC vs. ST inhibited ADP and AA-induced platelet aggregation significantly with a concentration of 40.7 μM.

Discussion and conclusions: As the processed product of ST, STC showed strong haemostatic activity on bleeding rat through regulating the parameters involved in haemorheology and plasma coagulation system. Two active compounds, dihydrocaffeic acid and amentoflavone, might be partially responsible for the haemostatic and anticoagulant activity of STC and ST, respectively.

Keywords: Haemorheology and plasma coagulation system; activity transformation; amentoflavone; dihydrocaffeic acid; natural products.

Publication types

  • Comparative Study

MeSH terms

  • Animals
  • Bleeding Time / methods
  • Blood Coagulation / drug effects*
  • Blood Coagulation / physiology
  • Blood Coagulation Tests / methods
  • Hemostatics / isolation & purification
  • Hemostatics / pharmacology*
  • Hot Temperature / adverse effects*
  • Male
  • Plant Extracts / isolation & purification
  • Plant Extracts / pharmacology*
  • Random Allocation
  • Rats
  • Rats, Sprague-Dawley
  • Selaginellaceae*

Substances

  • Hemostatics
  • Plant Extracts

Grants and funding

This work was supported by the National Natural Science Foundation of China (21275169 and 81202886), the Natural Science Foundation Project of CQ CSTC (cstc2015jcyjA10044). We sincerely acknowledge the Science and Technology Development Fund of Macao SAR and University of Macau for financial support by projects, FDCT013-2015-A1 and MYRG2016-00144-ICMS-QRCM for this research work.