Dataset on effect of thymol against influenza A virus in A549 cells

Published: 22 June 2022| Version 1 | DOI: 10.17632/kccn9d4htz.1
Contributors:
,
Madhu Khanna

Description

Influenza A virus, enveloped, segmented, negative strand RNA virus belongs to the family of Orthomyxoviridae, a major respiratory pathogen and responsible for causing number of global and local outbreaks. Currently, there are three classes of anti-influenza drugs with different antiviral mechanisms, including M2 ion channel blockers (amantadine and rimantadine), viral RNA synthesis inhibitors. However, evolution of NA inhibitor-resistant influenza viruses have also emerged, thus limiting the future utility of NA inhibitors. Thus the objective was to explore a low-toxic anti-influenza drug. The cytotoxicity assay of thymol was performed by MTT assay. Varying concentration of thymol ranging from 100 µg/ml to 0.39 µg/ml was added to A549 cells and absorbance was recorded at 570nm after 24hr. The data obtained was expressed in percentages. Based on the data, IC50 obtained was 49.5 µg/ml. Hence, concentrations from 50 µg/ml to 6.25 µg/ml was used for all the experiments. Thymol was evaluated against influenza A virus by real time PCR . RNA was isolated post infection and treatment and relative expression was measured by qPCR. The Ct value obtained was normalized against GAPDH. The real time PCR showed significant down regulation of IAV at 50 and 25 µg/ml. To further validate the anti-viral efficacy, the cells were further subjected to western blotting after treatment with thymol and infection with influenza A virus. Cellular protein was isolated and separated using 12 % SDS PAGE. The blot was probed using anti-PB1 influenza antibody. The expression was detected using chemiluminescence detection kit and chemidoc imaging system. The band intensity was quantified and plotted on a graph to measure the difference in expression of PB1 in both thymol treated and uninfected cells. The western blotting results showed significant inhibition of viral protein PB1 at 50 and 25 µg/ml, which corroborates with the real time PCR data. Thus, confirming the anti-viral efficacy of thymol against influenza A virus PR8 at 24hr in A549 cells. The graphs were statistically analyzed using One-Way analysis of variance (ANOVA). The data obtained might be useful in initiating in-vivo studies for establishing thymol as an effective treatment option against influenza A virus.

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