Cytotoxicity of Aconitum alkaloid and its interaction with calf thymus DNA by multi-spectroscopic techniques

Fei Liu,Nan Li,Xiaoxin Tan,Xu Han,Weijun Kang,Xiang Li

doi:10.1038/s41598-017-15240-9

Abstract

The cytotoxicities of three aconitum alkaloids- aconitine, hypaconitine and mesaconitine, and their abilities to bind DNA have been explored. Rat myocardial cells H9c2 were treated with aconitum alkaloids and assessed the cytotoxicities by using MTT assay and flow cytometry. Apoptosis was evidenced by the results of the annexin V/propidium iodide (PI) assay. Aconitine was found to be the most toxic in rat myocardial cells H9c2 in three aconitum alkaloids. At the same time, DNA adducts were isolated and then analyzed by UV-Vis spectroscopy after exposure to alkaloids, which indicated that three alkaloids could bind to DNA in rat myocardial cells H9c2. Furthermore, their binding modes were investigated by UV-Visible, fluorescence, DNA melting studies and ionic strength effect. Results indicated that the interaction between three alkaloids and DNA were intercalation coupled with electrostatic effect. The estimated binding constants were between 4.83 × 105 M−1 to 9.85 × 105 M−1 for three alkaloids at 298 K.

Highlights

The cytotoxicities of three aconitum alkaloids- aconitine, hypaconitine and mesaconitine, and their abilities to bind DNA have been explored
As we know, different Aconitum sources contain varies alkaloid: MA is the main compound of Aconitum kusnezoffii, AC is the main alkaloid in Aconitum napellus, and the main constituents of Aconitum carmichaeli are HA and MA
The LD50 values of hypaconitine and mesaconitine were significantly higher than aconitine, three of the alkaloids are all held responsible for the toxic effects

Summary

Introduction

The cytotoxicities of three aconitum alkaloids- aconitine, hypaconitine and mesaconitine, and their abilities to bind DNA have been explored. Aconitine was found to be the most toxic in rat myocardial cells H9c2 in three aconitum alkaloids. DNA adducts were isolated and analyzed by UV-Vis spectroscopy after exposure to alkaloids, which indicated that three alkaloids could bind to DNA in rat myocardial cells H9c2. Their binding modes were investigated by UV-Visible, fluorescence, DNA melting studies and ionic strength effect. Unprocessed Aconitum contains highly toxic alkaloids with a diester diterpene structure such as hypaconitine (HA), aconitine (AC), and mesaconitine (MA) (Fig. 1)[5]. DNA-adducts need to be examined using UV-Visible spectroscopy after exposure to various concentrations of DDAs

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