Biological activity of commercial medicinal Lamiaceae plant extracts from Serbia

Čovekova potraga za terapeutskim sredstvima prirodnog, a posebno biljnog porekla, datira još iz starih vremena. Sa napretkom savremene nauke omogućeno nam je da detaljno istražujemo njihove fitokonstituente, biološku aktivnost i mehanizme delovanja, sa ciljem dobijanja bezbednijih i efikasnijih farmakoloških agenasa. Cilj ove doktorske disertacije je bio određivanje hemijskog sastava i biološke aktivnosti metanolnih, etanolnih i vodenih ekstrakata 18 vrsta familije Lamiaceae koje se tradicionalno upotrebljavaju i komercijalno gaje u Srbiji. Hemijska karakterizacija je urađena korišćenjem spektrofotometrijskih metoda (određivanjem ukupnog sadržaja fenola, fenolnih kiselina, flavonoida, flavonola, kumarina i triterpena), kao i HPLC-DAD analizom. Biološki potencijal pomenutih ekstrakata je određen testiranjem njihove antioksidativne, genoprotektivne, antigenotoksične, antidijabetične, antineurodegenerativne, antineuroinflamatorne, antitumorske i antibakterijske aktivnosti, kao i procenom njihovog uticaja na modelu zarastanja rane. Ekstrakti ispitivanih predstavnika familije Lamiaceae obiluju sekundarnim metabolitima, posebno onima iz grupe fenolnih jedinjenja, pri čemu sastav i količina komponenti zavisi od primenjenog rastvarača za ekstrakciju. U ovim ekstraktima, najprisutnija fenolna kiselina je bila rozmarinska kiselina, a najprisutniji flavonoid, flavon luteolin-7-O-glukozid. Iako su svi ekstrakti ispoljili antioksidativnu aktivnost, najjači potencijal su imali ekstrakti Origanum vulgare i Melissa officinalis. Vodeni ekstrakti su pružali najviši nivo zaštite plazmidne DNK, posebno ekstrakti Mentha × piperita i Thymus vulgaris. Dok su najjaču antigenotoksičnu aktivnost u prokariotskom modelu imali etanolni ekstrakti Rosmarinus officinalis i Salvia officinalis, na eukariotskom modelu posebno dobru aktivnost su pokazali ekstrakti Hyssopus officinalis, Leonurus cardiaca i Marrubium vulgare. Iako ekstrakti nisu inhibirali aktivnost enzima α-amilaze, svoju antidijabetičnu aktivnost su ispoljili inhibicijom aktivnosti α-glukozidaze, pri čemu su se posebno izdvojili vodeni ekstrakti M. piperita, Satureja montana i Thymus serpyllum. Ispitivani ekstrakti imaju dobar antineurodegenerativni potencijal procenjivan inhibitornim efektom na aktivnost acetilholinesteraze tirozinaze, kao i antineuroinflamatorni i antineurocitotoksični efekat prema LPS-om aktiviranim ćelijama mikroglije. Većina ispitivanih etanolnih ekstrakata je imala antitumorski efekat prema ćelijskoj liniji kolorektalnog kancera. Genotoksični potencijal prema ovim tumorskim ćelijama koji je bio sličan efektu etopozida je ispoljio jedino etanolni ekstrakt Ocimum basilicum. Ekstrakti su imali i antibakterijski potencijal: inhibirali su rast testiranih bakterijskih sojeva, posebno gram-pozitivnih, inhibirali su invaziju Pseudomonas aeruginosa PAO1 prilikom infekcije fibroblasta pluća i formiranje biofilma P. aeruginosa PAO1, a takođe su degradirali i već formirani biofilm P. aeruginosa PAO1, pri čemu su se posebno izdvojili ekstrakti S. montana i O. vulgare. U ovoj doktorskoj disertaciji je po prvi put testirana i potvrđena genoprotektivna, kao i antigenotoksična aktivnost ispitivanih ekstrakata, a takođe po prvi put daje uvid u antineuroinflamatornu i antitumorsku aktivnost pojedinih predstavnika familije Lamiaceae. Na osnovu IBR analize svih ekstrakata predstavnika familije Lamiaceae, najbolji ukupni biološki potencijal su imali O. vulgare i M. officinalis, kao i etanolni ekstrakt S. officinalis, dok su najniži biološki potencijal pokazali ekstrakti M. vulgare, kao i etanolni ekstrakt G. hederacea.The man's search for natural therapeutic agents, especially those of plant origin, dates back to ancient times. However, with the progress of modern science, we have been able to investigate in detail their phytoconstituents, biological activity and mechanisms of action in the human body, in order to obtain safer and more effective farmaceuticals. The aim of this dissertation was to determine the chemical composition and biological activity of methanolic, ethanolic and aqueous extracts of 18 species of the Lamiaceae family that are being traditionally used and commercially grown in Serbia. Chemical characterization was performed using spectrophotometric methods (by the determination of the total content of phenols, phenolic acids, flavonoids, flavonols, coumarins, and triterpenes), as well as by HPLC-DAD analysis. The biological potential of the extracts was determined by testing their antioxidant, genoprotective, antigenotoxic, antidiabetic, antineurodegenerative, antineuro-inflammatory, antitumor, and antibacterial activities, as well as by assessing their influence on wound healing. The extracts of the tested Lamiaceae representatives are rich in secondary metabolites, mainly phenolic compounds. However, the content and amount of phytocomponents depends on the extraction solvent. In these extracts, the dominant phenolic acid was rosmarinic acid, and the dominant flavonoid was flavone luteolin-7-O-glucoside. Although all extracts showed antioxidant activity, Origanum vulgare and Melissa officinalis had the highest antioxidant potential. Aqueous extracts provided the highest plasmid DNA protection, especially Mentha × piperita and Thymus vulgaris. While the ethanolic extracts of Rosmarinus officinalis and Salvia officinalis had the strongest antigenotoxic activity in the prokaryotic model, Hyssopus officinalis, Leonurus cardiaca, and Marrubium vulgare extracts showed excellent activity in the eukaryotic model. Although the extracts had no effect on α- amylase inhibition, they exhibited antidiabetic activity by inhibiting the activity of α-glucosidase, especially aqueous extracts of M. piperita, Satureja montana and Thymus serpyllum. The tested extracts possess antineurodegenerative potential, which they exhibit by inhibiting the activities of acetylcholinesterase and tyrosinase, as well as antineuroinflammatory and antineurocytotoxic potentials against LPS-activated microglia. Most of the ethanolic extracts had an antitumor effect on colorectal cancer cells, while only the genotoxic effects of Ocimum basilicum were similar to etoposide. The extracts, especially S. montana and O. vulgare, exhibited their antibacterial potential by inhibiting the growth of tested bacterial strains, especially gram-positive ones, by inhibiting Pseudomonas aeruginosa PAO1 invasion during human lung fibroblast infection, as well as biofilm formation inhibition and degradation of already formed P. aeruginosa PAO1 biofilm. In this dissertation, the genoprotective and antigenotoxic activities of the examined plant extracts were tested and confirmed for the first time. Moreover, this study provides the first insight into the antineuroinflammatory and antitumor activities of certain investigated Lamiaceae representatives. Based on the IBR analysis of all tested Lamiaceae extracts, O. vulgare and M. officinalis, as well as ethanolic extract of S. officinalis had the best overall biological potential, while extracts of M. vulgare and ethanolic extract of G. hederacea showed the lowest biological potential