Elsevier

Biomedicine & Pharmacotherapy

Volume 87, March 2017, Pages 698-704
Biomedicine & Pharmacotherapy

Research Articles
The hemostatic effect study of Cirsium setosum on regulating α1-ARs via mediating norepinephrine synthesis by enzyme catalysis

https://doi.org/10.1016/j.biopha.2017.01.022Get rights and content

Abstract

Background

Cirsium setosum (CS) is the aboveground part of Cephalanoplos segetum Kitam. Although it has been used as a hemostatic treatment for thousands of years and is still in use today, the mechanism of CS on regulating ARs is still not clear.

Purpose

In this study, we aimed to clarify the mechanism of CS on regulating ARs.

Methods

We developed a simple method based on UPLC/Q-TOF MS combined adrenergic receptor dual-luciferase reporter assay systems for the rapid determination of active constituents in CS. The mechanism of tyramine, the main active component for regulating ARs, was further investigated by an in vitro norepinephrine biotransformation test and in vivo vaso activity tests.

Results

Two phenethylamine ARs regulators (tyramine and N-methyltyramine) in CS were characterized, and it was found that tyramine could induce vasoconstriction via regulation of α1-ARs by mediating norepinephrine synthesis.

Conclusion

The hemostatic effect of CS is associated with tyramine and N-methyltyramine, via regulation of α1-ARs, and the mechanism of tyramine is related to mediating norepinephrine synthesis by enzyme catalysis.

Introduction

Cirsium setosum (CS) is the aboveground part of Cephalanoplos segetum Kitam, which has been used as a traditional medicine against a variety of hemorrhagic diseases for thousands of years in China [1]. Currently, CS is commonly used to treat many types of diseases, including digestive tract bleeding, incomplete uterine contraction and bleeding, infectious hepatitis, and cardiovascular disease. Modern pharmacological studies have indicated that the active ingredients of CS have strong biological activity. Specifically, caffeic acid and chlorogenic acid in CS could promote aggregation and adhesion of platelets[2], [3], and rutin could reduce vascular permeability [4]. CS could play this role through multiple mechanisms, such as antihemorrhagic [5], anti-inflammatory [6], antimicrobial, and sedative functions, as well as enhancement of strong heart function [7], [8].

Additional research has shown that water decoction of CS had the effect of contracting rabbit aortic strips by activating α-AR and had an effect of relaxation of guinea pig trachea tablets by activation of β-AR [7]. It is well known that adrenergic receptors (ARs) are involved in regulating a series of important biological activities, including breath movement and regulation of myocardial and vessel contractility and that AR agonists are mainly phenethylamine alkaloids [9], [10]. These phenomena indicate that CS might play an important pharmacological role due to its relationship with ARs.

In this paper, first, an approach combining UPLC/Q-TOF MS and dual luciferase reporter assay for screening AR agonists was established and used to screen and identify active ingredients on ARs of total alkaloids in CS. Second, tyramine as a key active component was screened out from CS and was further investigated for hemostatic function. Finally, the mechanism of vasoconstriction was established by in vitro noradrenalin biotransformation tests and in vivo vasoactivity tests.

Section snippets

Samples preparation

One hundred grams of CS (purchased from Anguo herb market in the Hebei province of China and identified by Professor Tiejun Zhang from the Tianjin Institute of Pharmaceutical Research) raw material was soaked in 0.02 mol/L HCl overnight, and then supernatant was collected and filtered (WEX). The supernatant was passed through 001 × 7 cation exchange resin enrichment, washed with water until the effluent became neutral, and eluted with 5% of the ammonia water. Then, the eluted liquid (ENR) was

Effects of the total alkaloids of CS on AR activity

There are many chemical constituents included in total alkaloids of CS; therefore, to knock out weak activity components prior to further screening research we first experimented with the β2AR agonist activity of the different polarity extracts of total alkaloids in CS by the cellular assay. The results are shown in Fig. 1. The WEX, the extract of CS by 0.02 mol/L HCl, had a significant regulating action on β2AR, and its activity increased after being enriched by cation exchange resin. Among the

Conclusion

In this study, we found that the effect of CS is related to tyramine and N-methyltyramine via regulating α1-ARs, and the mechanism of tyramine is related to mediating noradrenalin synthesis by a series of enzyme catalysis.

Conflict of interest

The authors declare no conflict of interests.

Acknowledgements

This work was supported by a grant from the National Natural Science Foundation of China (nos. 81473403, 81373506), and the Natural Science Foundation of Tianjin, China (nos. 15JCYBJC28800).

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