title: 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.
abstract: 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, HeLa-CEACAM1a (HeLa-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 BioMark digital array software. Determinations of relative gene expression values were carried out by using the 2(-∆∆Ct) 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 8 hpi. 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).
authors.affiliation: Department of Medical Biology, Faculty of Medicine, University of Gaziantep, Şehitkamil, 27310, Gaziantep, Turkey, mulasli@gmail.com.
authors.forename: Mustafa
authors.initials: M
authors.lastname: Ulasli
authors.forename: Serdar A
authors.initials: SA
authors.lastname: Gurses
authors.forename: Recep
authors.initials: R
authors.lastname: Bayraktar
authors.forename: Onder
authors.initials: O
authors.lastname: Yumrutas
authors.forename: Serdar
authors.initials: S
authors.lastname: Oztuzcu
authors.forename: Mehri
authors.initials: M
authors.lastname: Igci
authors.forename: Yusuf Ziya
authors.initials: YZ
authors.lastname: Igci
authors.forename: Ecir Ali
authors.initials: EA
authors.lastname: Cakmak
authors.forename: Ahmet
authors.initials: A
authors.lastname: Arslan
mesh_terms: D000818:Animals; D031166:Anthemis / Q000737:chemistry; D032084:Citrus sinensis / Q000737:chemistry; D017934:Coronavirus / Q000187:drug effects* / Q000254:growth & development; D018352:Coronavirus Infections / Q000188:drug therapy* / Q000473:pathology / Q000821:virology; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics; D008519:Medicine, Traditional; D051379:Mice; D031881:Nigella sativa / Q000737:chemistry; D010936:Plant Extracts / Q000008:administration & dosage* / Q000737:chemistry; D050051:Transient Receptor Potential Channels / Q000096:biosynthesis*; D014779:Virus Replication / Q000187:drug effects
chemical_list: D016209:Interleukin-8; D010936:Plant Extracts; D050051:Transient Receptor Potential Channels