In vitro Antiviral Activity of Rubia cordifolia Aerial Part Extract against Rotavirus.

Yuanyuan Sun,Guang Du,Lifeng Kang,Jihong Yang,Jia Y. Tan,Xuepeng Gong,Dongyan Li,Vikash Vikash

doi:10.3389/fphar.2016.00308

Abstract

The root of Rubia cordifolia has been used traditionally as a hemostatic agent, while the aerial part of the plant consisting of leaf and stem is known to exhibit anti-diarrheal properties and has been widely used as a remedy in many parts of China. As rotavirus is one of the most commonly associated diarrhea-causing pathogen, this study aims to investigate the anti-rotaviral effect of R. cordifolia aerial part (RCAP). The cytotoxicity of RCAP toward MA-104 cells was evaluated using the WST-8 assay. Colloidal gold method and real time polymerase chain reaction (qPCR) assay were used to confirm the findings of the antiviral assay. Then, 4′,6-diamidino-2-phenylindole (DAPI) staining method was subsequently used to investigate the mode of death among the cells. And the representative components of aqueous extract were isolated and identified. It was shown that both the viability of MA-104 cells and the viral load were reduced with increasing concentration of the extract. DAPI staining showed that virus-induced apoptosis was the cause of the low cell viability and viral load, an effect which was accelerated with incubation in the aqueous herbal extract. The major compounds postulated to exhibit this activity were isolated from the aqueous herbal extract and identified to be compounds Xanthopurpurin and Vanillic Acid. This study showed that RCAP extract effectively inhibited rotavirus multiplication by promoting virus-induced apoptosis in MA-104 cells.

Highlights

Rotavirus, pathogenic to both man and animals, is a non-enveloped virus with a triple-layered double capsid and 11 segments of double-stranded Ribonucleic acid (RNA; Goncalves et al, 2005)
The morphological changes of the virus infected or non-infected R. cordifolia aerial part (RCAP) extract treated MA-104 cells were observed with light microscopy, after the cells in the virus control group (VCG, cells were only inoculated with virus) showed viral cytopathic effect (CPE)
It was found that these affected cells increased with an increasing concentration of the extract (Figure 2B), while the morphology of non-infected cells treated with RCAP extract remained consistent regardless of the RCAP extract concentration (Figure 2A)

Summary

Introduction

Pathogenic to both man and animals, is a non-enveloped virus with a triple-layered double capsid and 11 segments of double-stranded Ribonucleic acid (RNA; Goncalves et al, 2005). It is the most common diarrhea-causing pathogen in children world-wide (Parashar et al, 1998; Cecilio et al, 2012). Two million children are hospitalized, and 800,000 deaths occur due to rotavirus-associated diseases (Atherly et al, 2009; Mameli et al, 2012). Clinical manifestations of rotavirus gastroenteritis are typically fever, vomiting, abdominal pain, diarrhea and dehydration (Walker-Smith, 1978; Cecilio et al, 2012; Paul et al, 2014).

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