๐ The effects of Nigella sativa (Ns), Anthemis hyalina (Ah) and Citrus sinensis (Cs) extracts on the replication of coronavirus and the expression of TRP genes family
Extracts of Anthemis hyalina (Ah), Nigella sativa (Ns) and peels of Citrus sinensis (Cs) have been used as folk medicine to fight antimicrobial diseases. To evaluate the effect of extracts of Ah, Ns and Cs on the replication of coronavirus (CoV) and on the expression of TRP genes during coronavirus infection, He. La-CEACAM1a (He. La-epithelial carcinoembryonic antigen-related cell adhesion molecule 1a) cells were inoculated with MHV-A59 (mouse hepatitis virus-A59) at moi of 30. 1/50 dilution of the extracts was found to be the safe active dose. ELISA kits were used to detect the human IL-8 levels. Total RNA was isolated from the infected cells and cDNA was synthesized. Fluidigm Dynamic Array nanofluidic chip 96.96 was used to analyze the mRNA expression of 21 TRP genes and two control genes. Data was analyzed using the Bio. Mark digital array software. Determinations of relative gene expression values were carried out by using the 2-ฮฮCt method (normalized threshold cycle (Ct) value of sample minus normalized Ct value of control). TCID50/ml (tissue culture infectious dose that will produce cytopathic effect in 50 % of the inoculated tissue culture cells) was found for treatments to determine the viral loads. The inflammatory cytokine IL-8 level was found to increase for both 24 and 48 h time points following Ns extract treatment. TRPA1, TRPC4, TRPM6, TRPM7, TRPM8 and TRPV4 were the genes which expression levels changed significantly after Ah, Ns or Cs extract treatments. The virus load decreased when any of the Ah, Ns or Cs extracts was added to the CoV infected cells with Ah extract treatment leading to undetectable virus load for both 6 and 8hpi. Although all the extract treatments had an effect on IL-8 secretion, TRP gene expression and virus load after CoV infection, it was the Ah extract treatment that showed the biggest difference in virus load. Therefore Ah extract is the best candidate in our hands that contains potential treatment molecule(s).
keywords
๐ cytopathic effect (91)
๐ tissue culture (67)
๐ adhesion molecule (17)
๐ hepatitis virus (437)
๐ mouse hepatitis (371)
๐ coronavirus infection (270)
๐ infected cells (307)
๐ viral load (91)
author
๐ค Ulasli, Mustafa
๐ค Gurses, Serdar A.
๐ค Bayraktar, Recep
๐ค Yumrutas, Onder
๐ค Oztuzcu, Serdar
๐ค Igci, Mehri
๐ค Igci, Yusuf Ziya
๐ค Cakmak, Ecir Ali
๐ค Arslan, Ahmet
year
โฐ 2014
journal
๐ Molecular Biology Reports
issn
๐ 15734978 03014851
volume
41
number
3
page
1703-1711
citedbycount
3
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