Yildirim Ayse, Duran Gulay Gulbol, Duran Nizami, Jenedi Kemal, Bolgul Behiye Sezgin, Miraloglu Meral, Muz Mustafa
Department of Histology and Embryology, Mustafa Kemal University, Medical Faculty, Hatay, Turkey.
Department of Medical Biology, Mustafa Kemal University, Medical Faculty, Hatay, Turkey.
Med Sci Monit. 2016 Feb 9;22:422-30. doi: 10.12659/msm.897282.
BACKGROUND Propolis is a bee product widely used in folk medicine and possessing many pharmacological properties. In this study we aimed to investigate: i) the antiviral activities of Hatay propolis samples against HSV-1 and HSV-2 in HEp-2 cell line, and ii) the presence of the synergistic effects of propolis with acyclovir against these viruses. MATERIAL AND METHODS All experiments were carried out in HEp-2 cell cultures. Proliferation assays were performed in 24-well flat bottom microplates. We inoculated 1x105 cells per ml and RPMI 1640 medium with 10% fetal calf serum into each well. Studies to determine cytotoxic effect were performed. To investigate the presence of antiviral activity of propolis samples, different concentrations of propolis (3200, 1600, 800, 400, 200, 100, 75, 50, and 25 μg/mL) were added into the culture medium. The amplifications of HSV-1 and HSV-2 DNA were performed by real-time PCR method. Acyclovir (Sigma, USA) was chosen as a positive control. Cell morphology was evaluated by scanning electron microscopy (SEM). RESULTS The replication of HSV-1 and HSV-2 was significantly suppressed in the presence of 25, 50, and 100 μg/mL of Hatay propolis. We found that propolis began to inhibit HSV-1 replication after 24 h of incubation and propolis activity against HSV-2 was found to start at 48 h following incubation. The activity of propolis against both HSV-1 and HSV-2 was confirmed by a significant decrease in the number of viral copies. CONCLUSIONS We determined that Hatay propolis samples have important antiviral effects compared with acyclovir. In particular, the synergy produced by antiviral activity of propolis and acyclovir combined had a stronger effect against HSV-1 and HSV-2 than acyclovir alone.
蜂胶是一种广泛应用于民间医学且具有多种药理特性的蜂产品。在本研究中,我们旨在调查:i)哈塔伊蜂胶样品在人喉表皮样癌细胞系(HEp-2)中对单纯疱疹病毒1型(HSV-1)和单纯疱疹病毒2型(HSV-2)的抗病毒活性,以及ii)蜂胶与阿昔洛韦联合对这些病毒的协同作用。
所有实验均在HEp-2细胞培养物中进行。增殖试验在24孔平底微孔板中进行。我们向每孔接种每毫升1×10⁵个细胞以及含有10%胎牛血清的RPMI 1640培养基。进行了确定细胞毒性作用的研究。为了研究蜂胶样品的抗病毒活性,向培养基中添加了不同浓度的蜂胶(3200、1600、800、400、200、100、75、50和25μg/mL)。通过实时聚合酶链反应(PCR)方法进行HSV-1和HSV-2 DNA的扩增。选用阿昔洛韦(美国西格玛公司)作为阳性对照。通过扫描电子显微镜(SEM)评估细胞形态。
在存在25、50和100μg/mL哈塔伊蜂胶的情况下,HSV-1和HSV-2的复制受到显著抑制。我们发现,孵育24小时后蜂胶开始抑制HSV-1复制,而蜂胶对HSV-2的活性在孵育48小时后开始显现。病毒拷贝数的显著减少证实了蜂胶对HSV-1和HSV-2两者均有活性。
我们确定,与阿昔洛韦相比,哈塔伊蜂胶样品具有重要的抗病毒作用。特别是,蜂胶和阿昔洛韦联合产生的抗病毒活性协同作用对HSV-1和HSV-2的效果比单独使用阿昔洛韦更强。