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Methanolic Extract of Neem Plant Inhibits NS3 and NS5A Nonstructural Proteins of HCV 3A Genotype

Methanolic Extract of Neem Plant Inhibits NS3 and NS5A Nonstructural Proteins of HCV 3A Genotype

Ubaida Hussain1,2*, Amtul Jamil Sami1*, Shazia Rafique3*, Muhammad Idrees khan3 and Ahmad Ali Shahid3

1School of Biochemistry and Biotechnology University of the Punjab, Quaid-I-Azam Campus, Lahore 54590, Pakistan
2School of Chemistry, University of the Punjab, Quaid-I-Azam Campus, Lahore 54590, Pakistan
3Center for Excellence in Molecular Biology, University of the Punjab, West Canal Bank Road Thokar Niaz Baig, Lahore 53700, Pakistan
 
* Corresponding author: ubaida.chem@pu.edu.pk, amtul.ibb@pu.edu.pk,shazia.rafique@cemb.edu.pk

ABSTRACT

Hepatitis C virus (HCV) has infected an approximate of 170-200 million people worldwide. This unique RNA virus escapes the immunity in most infected people and forms a chronic infection which can progress to liver cirrhosis, fibrosis, hepatic cancer, and death. The goal of this work was to assess the cytotoxicity and in vitro inhibitory activities of local medicinal plant Azadirachta indica (A. indica) extracts on expression of NS3 and NS5A nonstructural proteins of HCV 3a genotype. Owing to the fact that HCV patients can develop HCC, plant extract has also been tested for cytotoxic activity. Colorimetric analysis was done to determine the vitality of HepG 2 cells for 24 h after treatment with methanolic seed extract and minimum inhibitory concentration was calculated. HepG2 cell were transfected stably to express nonstructural proteins NS3 and NS5A of HCV 3a genotype. Western blot and RT PCR confirmed stable protein expression. Immunofluorescence assay was done to check subcellular localization of these nonstructural proteins. The CC50 value for A. indica seed extract for HepG2 cells was 650 μg/mL. The extract concentration 500 μg/mL was found to reduce gene and protein expression of NS3 and NS5A by 50 percent. Results of our study shows that phytochemicals of neem seeds have strong antiviral potential against HCV without having any significant cytotoxic effects.

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Pakistan Journal of Zoology

June

Pakistan J. Zool., Vol. 56, Iss. 3, pp. 1001-1500

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