Antimikrobni i citotoksični potencijal etanolnih ekstrakata odabranih predstavnika podfamilije Polygonoideae i njihov efekat u kombinaciji sa streptomicinom i doksorubicinom

Abstract

Za ispitivanje antimikrobnog i citotoksičnog efekta odabrano je 8 biljaka podfamilije Polygonoideae od kojih je napravljeno 12 etanolnih ekstrakata, od herbi (h) i rizoma (r): Polygonum aviculare (POAh, POAr), Polygonum maritimum (POMh), Persicaria amphibia (PEAh), Persicaria maculosa (PEMh), Persicaria lapathifolia (PELh, PELr), Persicaria hydropiper (PEHh, PEHr), Bistorta officinalis (BIOh, BIOr) i Fagopyrum esculentum (FAEh). Svi testirani ekstrakti ispoljavaju umerenu antifungalnu aktivnost, a POAh, POMh, PEAh, POAr, PEHr i BIOr dovode do obezbojavanja konidija. Antibakterijski efekat je testiran prema većem broju G+ i G- bakterija pri čemu je BIOr pokazao najpotentnije antimikrobno dejstvo. BIOr i PEMh pokazuju značajan antibiofilm i antiquorum sensing efekat prema medicinski važnom patogenu Pseudomonas aeruginosa PAO1 i to preko modulacije aktivnosti LasR receptora. Ovi ekstrakti i njihovi aktivni konstituenti, hinska i hlorogenska kiselina, katehin i kvercetin- 3-O-glukozid, inhibiraju sintezu violaceina kod Chromobacterium violaceum. U kombinaciji sa streptomicinom BIOr ostvaruje različite tipove interakcija: antagonistički, indiferentni, aditivni i sinergistički. Prilikom testiranja citotoksičnosti odabranih ekstrakata i doksorubicina (D) uočen je sinergizam između P. aviculare i D na ćelijama hepatocelularnog adenokarcinoma (HepG2) i plućnog adenokarcinoma (A549). Takođe, sinergizam između P. amphibia i D je detektovan na A549 ćelijama. Sinergistički efekat se ostvaruje preko sledećih procesa: uvođenje ćelija u apoptozu, zaustavljanje ćelijskog ciklusa, modelovanje aktivnosti antioksidativnih enzima i Keap1-Nrf2 signalnog puta. Na normalnim ćelijama fibroblasta pluća (MRC-5) P. aviculare i D, P. amphibia i D kao i P. maculosa i D ostvaruju antagonizam. Odsustvo toksičnosti PEMh i BIOr u nižim dozama, potvrđeno je in vivo testom na modelu Danio rerio.

were selected and 12 ethanol extracts were made. Herb (h) and rhizome (r) extracts were obtained from: Polygonum aviculare (POAh, POAr), Polygonum maritimum (POMh), Persicaria amphibia (PEAh), Persicaria maculosa (PEMh), Persicaria lapathifolia (PELh, PELr), Persicaria hydropiper (PEHh, PEHr), Bistorta officinalis (BIOh, BIOr) and Fagopyrum esculentum (FAEh). All tested extracts moderately inhibited the growth of selected fungi. POAh, POMh, PEAh, POAr, PEHr and BIOr leads to discoloration of conidia. Antibacterial activity on different Gram positive and Gram negative strains has been evaluated. BIOr showed the most potent antibacterial activity. Also, BIOr and PEMh exhibited significant antibiofim and antiquorum sensing effects against medically important pathogen, Pseudomonas aeruginosa PAO1, through the modulation of LasR receptor activity. Both extracts as well as their active constituents, chinic acid, chlorogenic acid, catechin and quercetin-3-O-glucoside, inhibited violacein synthesis in Chromobacterium violaceum. The BIOr engaged in various interactions with streptomycin: synergic, addictive, indifferent and antagonistic. Cytotoxicity testing of selected extracts and doxorubicin (D) revealed a synergistic relationship between P. aviculare and D, in hepatocellular adenocarcinoma (HepG2) and pulmonary adenocarcinoma cells (A549). Also, synergism between P. amphibia and D, was detected on A549 cells. The observed synergisms could be linked to the interference with apoptosis, cell cycle, antioxidant enzymes expression and Keap1-Nrf2 pathway. On normal lung fibroblast cells (MRC-5) P. aviculare and D, P. amphibia and D as well as P. maculosa and D exerts an antagonistic effect. In vivo assay on the Danio rerio model shows no signs of toxicity at lower tested concentrations for PEMh and BIOr.