Abstract
Introduction. The abolition of smallpox vaccination after its elimination in 1980 led to a decrease in the immunocompromised immunity in humans. Zoonotic monkeypox, camelpox, buffalopox and cowpox viruses that are close to the variola virus also pose a danger to humans. In Russia today there are no effective and safe medicines for the prevention and treatment of smallpox and other orthopoxvirus infections in humans and animals. The Lady's mantle (Alchemilla vulgaris) is a promising source for the development of new antiviral drugs. Previous studies have found that Alchemilla vulgaris shows activity against influenza virus and herpes simplex virus.Aim. The aim of this work was to study the chemical composition and antiviral activity of extracts from the roots and the grass of Alchemilla vulgaris against orthopoxviruses.Materials and methods. Qualitative analysis of the samples was performed by high performance liquid chromatography. Quantitative analysis was performed using a complex of spectrophotometric methods. To determine the toxicity and antiviral activity of experimental samples from Alchemilla vulgaris in vitro, a transplantable Vero cell culture was used. Antiviral activity of the obtained preparations was evaluated by reducing the infectivity (titer) of orthopoxviruses in the monolayer of Vero cells infected with orthopoxviruses in the presence of preparations with different concentrations relative to the cell culture infected with orthopoxviruses without the preparations.Results and discussion. It was shown that the experimental sample from Alchemilla vulgaris obtained by the method of ethyl acetate extraction from the roots of the studied plant and purified with chloroform contains mainly catechins and leucoanthocyanins (70 %). In parallel, extract from the raw mass of the grass of the plant purified with chloroform and ethanol contains the amount of flavonoids (71 %). Wherein the content of flavonoids in unpurified ethanol extracts from the roots and the grass of Alchemilla vulgaris composed 5 % and 6 %, respectively. It was revealed that purified preparations obtained from Alchemilla vulgaris roots when using ethyl acetate and ethanol as extractants showed antiviral activity against vaccinia virus and ectromelia virus, as well as preparations obtained from grass by ethanol extraction.Conclusion. Thus, purified ethyl acetate extracts from the roots and ethanol extracts from the wet grass mass of Alchemilla vulgaris exhibit antiviral activity against orthopoxviruses in vitro.
Highlights
The abolition of smallpox vaccination after its elimination in 1980 led to a decrease in the immunocompromised immunity in humans
Previous studies have found that Alchemilla vulgaris shows activity against influenza virus and herpes simplex virus
Antiviral activity of the obtained preparations was evaluated by reducing the infectivity of orthopoxviruses in the monolayer of Vero cells infected with orthopoxviruses in the presence of preparations with different concentrations relative to the cell culture infected with orthopoxviruses without the preparations
Summary
В работе использовали измельченные корни и траву Alchemilla vulgaris, собранной на Семинском перевале Горного Алтая в фазу бутонизации – начала цветения в 2012 году. Выход этанольного экстракта для травы и корней Alchemilla vulgaris составляет 30 % и 35 % соответственно. Получение образцов на основе суммы флавонолов из травы Alchemilla vulgaris. Получение экспериментальных образцов препаратов на основе суммы флавоноидов (катехинов и лейкоантоцианов) из корней Alchemilla vulgaris. Определяли максимально переносимые концентрации (МПК) образцов для данной культуры клеток. После этого по 100 мкл разведений экстрактов вносили в соответствующие лунки планшета (по 4 лунки для каждой концентрации) и ставили в термостат на 4 суток при температуре 37 oC, 5 % СО2 и 100 % влажности. Определяли МПК, равную концентрации экстракта, при которой сохранялось 100 % жизнеспособного монослоя клеток во всех лунках. Через 6 суток культивирования в монослое клеток подсчитывали количество бляшек и определяли титр вирусов (в lg БОЕ/мл) в опыте и контроле (без препаратов).
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