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In Vitro Anti-Herpes Simplex Virus Activity, Antioxidant Potential and Total Phenolic Compounds of Selected Iranian Medicinal Plant Extracts

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In Vitro Anti-Herpes Simplex Virus Activity, Antioxidant Potential and Total Phenolic Compounds of Selected Iranian Medicinal Plant Extracts Indian Journal of Traditional Knowledge Vol. 17 (2), April 2018, pp 255-262 In vitro anti-herpes simplex virus activity, antioxidant potential and total phenolic compounds of selected Iranian medicinal plant extracts Mohammad-Taghi Moradi1, Ali Karimi1*, Somayeh Alidadi2 & Leila Hashemi3 1Medical Plants Research Center; 2Cellular and molecular Research Center; 3Clinical Biochemistry Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Science, Shahrekord, Iran E-mails: [email protected], [email protected] Received 11 September 2017, revised 17 January 2018 Drug resistant strains of herpes simplex virus-1 (HSV-1) have been increased recently. Essentially, medicinal plant-based new antiviral agents that are effective against HSV-1 infections are urgently required. Therefore, this research was conducted to evaluate in vitro anti HSV-1 activity of 25 plant extracts. In this study, the hydroalchoholic extracts of different parts of 25 medicinal plants belonging to 16 different families were prepared. Anti-HSV-1 activity was evaluated on Vero cell line using MTT (3-[4,5-dimethylthiazol–2-yl]-2,5-diphenyltetrazolium bromide) assay. The 50 % effective concentration (EC50) and 50 % cytotoxicity concentration (CC50) of the extract were determined using regression analysis. The inhibitory effect of the plant materials on adsorption and/or post-adsorption stages of HSV-1 replication cycle was determined. Antioxidant activity, total phenolic content, and total flavonoid content of the extracts were determined using 2, 2-diphenyl-1- picrylhydrazyl (DPPH) assay, Folin-Ciocalteu reagent, and aluminum chloride colorimetric method, respectively. Results showed that Pistacia atlantica Desf., Equisetum arvense L., Melissa officinalis L., Anthyllis vulneraria L., Punica granatum L., Syzygium aromaticum (L.) Merr. & L.M. Perry, Camellia sinensis (L.) Kuntze, and Crataegus azarolus L., were active against HSV-1. There was a significant association of total phenolic contents (R = 0.773, p < 0.001) and the free radical scavenging property (R = -0.684, p < 0.01) with the antiviral activity of the extracts. Some of the Iranian plant extracts studied showed potent antiviral activity against HSV-1 and can be used to develop new and effective anti-HSV-1 agents. Keywords: Antiviral activity, Herpes simplex virus, Antioxidant potential IPC Int. Cl.8: A61P31/12, C07K 14/035, A61K 39/245, C09K 15/00 Plants have the ability to synthesize a wide array of mild to severe life threatening infections such as compounds and have long been used as remedies, with encephalitis14. During the past two decades, the many of them being currently collected and examined mechanisms of replication and pathogenesis of HSV-1 to identify possible sources of herbal medicines1. and thus the potential antiviral targets in this virus have Some medicinal plant extracts have been recently used been thoroughly understood, leading to development of for treatment of a few diseases2 and some others antiviral compounds for this virus14. A category of have been evaluated to develop therapeutic agents with nucleoside analogues, acyclovir (ACV), is widely used promising results3-5. to treat HSV-1 infections15. However, a major problem Modern studies have shown that some of the with the use of ACV, is the development of drug medicinal plants with therapeutic application in resistant HSV strains, particularly in AIDS patients16. traditional medicine6-9 have antiviral effects. Many This type of drug resistance may occur after long-term screening efforts have been made to seek out antiviral treatment and is mainly due to mutations in the HSV-1 agents from natural sources10-13. thymidine kinase and/or DNA polymerase genes17. Persia ancient civilization has a long history in Therefore, antiviral, medicinal plant-based agents with identification of medicinal plants and treatment by active compounds that exhibiy different modes of these plants and Iranian scientists such as Avicenna action against viral infections are urgently needed. have tried to develop this science. Iran is endowed with rich and diverse local health Infections caused by HSV-1 represent a public tradition, which is matched with its rich and diverse health concern worldwide. These infections range from plant genetic source. Detailed investigations and —————— documentation of plants used in local healthcare *Corresponding author traditions and ethnopharmacological evaluation to 256 INDIAN J TRADIT KNOWLE, VOL. 17, NO. 2, APRIL 2018 verify their efficacy and safety can lead to the dissolved in 70 % ethylealcohole and kept at room development of invaluable herbal drugs or isolation temperature in an orbital shaker for 96 h. Next, the of compounds of therapeutic value. Therefore, this mixture was filtered and concentrated under research was conducted to examine 25 plant extracts approximately vacuum pressure and 40 °C in a for antiviral activity against HSV-1 in vitro. rotary evaporator. The extracts were kept in sterile bottles under refrigerated conditions until further Methodology use. The extracts were suspended at 37 °C in 10 % dimethylsulphoxide (DMSO) to yield a stock solution Plant collection and extraction of 10 mg/mL. This solution was filtered (Millipore® Different parts of 25 medicinal plants belonging to 0.22 μm) and stored (at 4 °C) until used. The small 16 different families (Table 1) that are traditionally percentage (up to 0.2 %) of DMSO present in the wells used for treatment of diseases were tested for their has no effect on the results of the experiments18. antiviral activity. The plant materials were provided from medicinal plants market in Shahrekord, a South Determination of free-radical scavenging activity west city of Iran. Then, both genus and species of the The free-radical scavenging activity was measured plants were identified and confirmed in the Herbarium by the 2, 2 diphenyl-1-picrylhydrazyl (DPPH) assay of Medical Plants Research Center of Shahrekord described by Moon & Terao, with some modification19. University of Medical Sciences. The plant materials Briefly, different amounts of the extracts and methanol were shadow dried, pulvarized, and then extracted were added to a solution of 0.3 mg/mL methanol using maceration method. The plant material was solution of DPPH to a final volume of 3.0 m. Table 1 — Ethnobotanical data of selected medicinal plants NO Botanical name (Family) Local name Voucher number Common name Parts used 1 Allium ampeloprasum L. (Amaryllidaceae) Tarrehkoei MPSKUMS-169 Wild leek Aerial parts 2 Allium jesdianum Boiss. (Amaryllidaceae) bosor MPSKUMS-204 - Leaves 3 Pistacia atlantica Desf. (Anacardiaceae) Baneh MPSKUMS-93 Atlas mastic tree Leaves 4 Peucedanum membranaceum Boiss (Apiaceae) Chowak MPSKUMS-57 Hog's fennel Aerial parts 5 Kelussia odoratissima Mozaff (Apiaceae) Kelos MPSKUMS-144 Wild celery Leaves 6 Heracleum persicum Desf. (Apiaceae) Golpar MPSKUMS-267 Golpar Fruits 7 Achilleam illefolium L. (Asteraceae) Boomadaran MPSKUMS-304 Yarrow Aerial parts 8 Calendula officinalis L. (Asteraceae) Hamyshebahar MPSKUMS-116 Marigold Aerial parts 9 Berberis integerrima Bunge (Berberidaceae) Zereshkekoohi MPSKUMS-512 Barberry root 10 Equisetum arvense L. (Equisetaceae) Domeasb MPSKUMS-516 Horsetail Aerial parts 11 Vaccinium arctostaphylos L. (Ericaceae) Ghareghat MPSKUMS-264 Whortleberry Fruits 12 Melissa officinalis L. Badranjbuoye MPSKUMS-107 lemon balm Aerial parts (Lamiaceae) 13 Stachys lavandulifolia Vahl (Lamiaceae) Chaeekoohi MPSKUMS-204 - flowers 14 Ziziphora clinopodioides Lam. (Lamiaceae) Kakooti MPSKUMS-253 - Aerial parts 15 Zataria multiflora Boiss. (Lamiaceae) Avishanshirazi MPSKUMS-443 - Aerial parts 16 Thymus daenensis Celak. (Lamiaceae) Avishandenaei MPSKUMS-248 thyme Aerial parts 17 Hyssopus officinalis L. (Lamiaceae) Zofa MPSKUMS-177 hyssop flowers 18 Trifolium campestre Schreb. (Leguminosae) Shabdarezard MPSKUMS-1001 Kidney vetch Leaves 19 Punica granatum L. Anar MPSKUMS-316 Pomegranate Peel fruits (Lythraceae) 20 Althaea officinalis L. Khatmi MPSKUMS-139 Marsh-mallow Flowers (Malvaceae) 21 Syzygium aromaticum (L.) Merr. & L.M. Perry Mikhak MPSKUMS-1002 Clove Flower buds (Myrtaceae) 22 Rheum ribes L. Rivas MPSKUMS-527 Currant-fruited Petiolules (Polygonaceae) rhubarb 23 Adiantum capillus-veneris L. (Pteridaceae) Parsiyavash MPSKUMS-131 maidenhair Leaves 24 Crataegus azarolus L. Zalzalak MPSKUMS-499 Hawthorn Leaves (Rosaceae) 25 Camellia sinensis (L.) Kuntze (Theaceae) Chaee MPSKUMS-1003 Black tea Leaves MORADI et al.: ANTI-VIRAL ACTIVITY OF IRANIAN PLANT EXTRACTS 257 After standing for 15 min at RT, the absorbance was grown in Dulbecco's Modified Eagle's Medium read at 517 nm using UV–Vis pectrophotometer (DMEM; Gibco, USA) supplemented with 10 % of (UNICO 2100: USA). High absorbance of the reaction fetal bovine serum (FBS) (Gibco, USA), 100 µg/mL of mixture indicated low free radical scavenging activity. streptomycin, 100 UI/mL of penicillin, and 0.25 µg/mL Butylated hydroxytoluene (BHT) was used as positive of amphotericin B (Gibco, USA) at 37 °C with 5 % control. Inhibition of free radical by DPPH was CO2. The same medium containing 2 % PBS was used calculated according to the formula below: for cytotoxicity and antiviral assays. HSV-1 (HSV-1, KOS strain) was kindly provided by University of Antiradical activity (%) = (Acontrol – Asample)/Acontrol × 100. Tarbiat Modares, Tehran, Iran. Virus stock was prepared by infection of confluent monolayer Vero The IC50 value, defined as the amount of antioxidant 2 necessary to decrease
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