For all the assays, ACV control consisted of the cells, ACV at different concentrations: 3.125, 6.25, 12.5, and 25 μg/ml for HSV-2 and 0.78, 1.56, 3.125, 6.25, and 12.5 μg/ml for HSV-1 made by 1:2 serial dilutions and the virus suspension. It was dissolved with DMSO to give a stock concentration of 10 mg/ml and then diluted with DMEM-2% FBS (maintenance medium) before use. HSV-1 clinical strain and HSV-2 ATCC G strain virus stocks were propagated in Vero cells and used at a titer of 1:10,000 for HSV-1 and 1:1000 for HSV-2 in all in-vitro experiments.Īcyclovir (ACV) powder (Matrix Laboratories Limited, Hyderabad, India) was used as a standard antiviral drug at four different concentrations: 3.125, 6.25, 12.5, and 25 μg/ml for HSV-2 and 0.78, 1.56, 3.125, 6.25, and 12.5 μg/ml for HSV-1. Confluent monolayer of African Green Monkey kidney cell line (Vero) (ATCC CCL-81) was grown in T-25 flasks in a 5% carbon dioxide (CO 2 ) incubator at 37☌ for 48 hours, in DMEM supplemented with 10% FBS, penicillin G sodium 200 U/ml, streptomycin sulfate 200 mg/L, and amphotericin B 0.5 mg/L. Table 1: Details of the plant and their extracts screened against herpes virusĪll reagents and media for cell culture were purchased from GIBCO-BRL (Grand Island, NY, USA). The final concentration of DMSO was <0.2%. 12430) supplemented with 2% fetal bovine serum (FBS) (GIBCO Cat. Louis, USA) and subsequent serial dilutions were carried out in Dulbecco's modified eagle medium (DMEM) (GIBCO Cat. The stock solution of 20 mg/ml was prepared in dimethyl sulfoxide (DMSO) (Sigma, St. After each cycle, solvent was added to replenish the remaining volume and dried in a rotatory evaporator at temperature not exceeding 35☌. Water and methanol extracts were prepared from 500 g of fresh raw material of plant parts in Soxhlet extraction vessel under reflux with stirrer for 3 cycles with methanol or water. The herb powders were prepared by drying and grinding the plant materials. We screened 24 extracts of 7 selected plants, of which 7 were water extracts, 3 were methanol extracts, and 14 were powders of various plant parts. The figure depicts the plants and the methodology used for screening įigure 1: In-vitro screening of plant extracts against HSV-1 and HSV-2įigure 2: Twenty-four extracts of seven plants were screened for their antiviral activity in-vitro. They were authenticated at the Botanical Survey of India, Pune and voucher specimens were deposited and standardized using High Performance Thin Layer Chromatography (HPTLC) method (data not provided here) and. The plant species were taxonomically identified and confirmed using morphological and anatomical techniques. sanctum, Nyctanthes arbortristis, Carica papaya, and Holarrhena antidysentrica were collected from Gujarat State of India. indica, Curcuma longa, Punica granatum, O. grantum have potential to be further explored for its possible anti-HSV activity. Thus, aqueous extract and freeze-dried powder of P. PGFP showed antiviral activity at 50, 100, and 200 μg/ml concentrations followed by PGAE, which showed highest selectivity index (SI) (14 and 12.5) followed by PGFP (7.6 and 12.9). Five extracts, Punica granatum aqueous extract (PGAE), peals powder (PGPP), freeze-dried powder (PGFP), Ocimum sanctum methanol extract (OSME), and Azadirachta indica leaves powder (AZLP) demonstrated favorable activity in primary screening. Primary screening of 24 extracts of seven plants was done by cytopathic effect inhibition (CPE) assay followed by dose response, antiviral, and cytotoxicity assay conducted at eight concentrations from 3.125 to 400 μg/ml. ,, ,, We investigated the plants selected on the basis of their traditional usage by comparing the disease symptomatology and recommended use from the texts of Ayurveda and used them in the forms as used traditionally. Treatment of herpes infection is thus cause of major concern owing to the difficulty in eliminating it from the ganglion, high cost of treatment, increasing drug resistance, and association with HIV-1. Primary infection within the genital tract, followed by an established latency phase give rise to life-long infection in humans. Herpes simplex virus 1 and 2 (HSV-1 and HSV-2) belong to diverse family of Herpesviridae, causing oral herpes lesions (HSV-1), genital lesions (HSV-2), meningitis, and encephalitis. Antiviral potential of selected Indian medicinal (ayurvedic) plants against Herpes Simplex Virus 1 and 2. How to cite this URL: Jadhav P, Kapoor N, Thomas B, Lal H, Kshirsagar N. How to cite this article: Jadhav P, Kapoor N, Thomas B, Lal H, Kshirsagar N.
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