Screening and Identification of Cardioprotective Compounds From Wenxin Keli by Activity Index Approach and in vivo Zebrafish Model.

Hao Liu,Yi Wang,Xiaoping Zhao,Xuechun Chen,Mirko Baruscotti,Yang Shi,Buchang Zhao,Ke Qian

doi:10.3389/fphar.2018.01288

Abstract

Wenxin Keli (WXKL) is a widely used Chinese botanical drug for the treatment of arrhythmia, which is consisted of four herbs and amber. In the present study, we analyzed the chemical composition of WXKL using liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) to tentatively identify 71 compounds. Through typical separate procession, the total extract of WXKL was divided into fractions for further bioassays. Cardiomyocytes and zebrafish larvae were applied for assessment. In vivo arrhythmia model in Cmlc2-GFP transgenic zebrafish was induced by terfenadine, which exhibited obvious reduction of heart rate and occurrence of atrioventricular block. Dynamic beating of heart was recorded by fluorescent microscope and sensitive camera to automatically recognize the rhythm of heartbeat in zebrafish larvae. By integrating the chemical information of WXKL and corresponding bioactivities of these fractions, activity index (AI) of each identified compound was calculated to screen potential active compounds. The results showed that dozens of compounds including ginsenoside Rg1, ginsenoside Re, notoginsenoside R1, lobetyolin, and lobetyolinin were contributed to cardioprotective effects of WXKL. The anti-arrhythmic activities of five compounds were further validated in larvae model and mature zebrafish by measuring electrocardiogram (ECG). Our findings provide a successful example for rapid discovery of bioactive compounds from traditional Chinese medicine (TCM) by activity index based approach coupled with in vivo zebrafish model.

Highlights

Natural products have played important roles in healthcare system throughout history and will continue to be served as huge and invaluable resource for the discovery of drug candidates
Twenty-seven compounds of them belong to Notoginseng, including notoginsenoside R1, ginsenoside Rg1, ginsenoside Re, ginsenoside Ra3, ginsenoside Rb1, notoginsenoside R2, ginsenoside Rc, ginsenoside Rd and so on
Wang et al established a database for the chemical components of the five herbs in Wenxin Keli (WXKL) for active compounds predication (Wang et al, 2017), all the compounds were acquired from database refer to the herb not the real composition of the patent drug, and the chemical components probably change during the manufactory process

Summary

Introduction

Natural products have played important roles in healthcare system throughout history and will continue to be served as huge and invaluable resource for the discovery of drug candidates. Traditional Chinese Medicine (TCM), widely used in Eastern Asian countries, has been regarded as an important part of natural products for the therapy of various diseases (Wang et al, 2012). As a consequence, developing high throughput methods with satisfied sensitivity for identifying active compounds from complex mixtures of TCM is in great demand. Many efforts have been made for rapidly screening of compounds from TCM. Artemisinin (qinghaosu) with antimalarial effect is a successful and impressive example as the gift from TCM (Tu, 2011)

Methods

Results

Conclusion