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2020, vol. 61, br. 1, str. 44-51
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In vitro ispitivanje antioksidativne aktivnosti preparata ruzmarinske kiseline i meda
In vitro antioxidant activity evaluation of rosmaric acid and honey based supplement
aHealthcare center "Sloga Medik", Kruševac bUniverzitet u Beogradu, Farmaceutski fakultet, Institut za medicinsku biohemiju cUniverzitet u Beogradu, Farmaceutski fakultet, Institut za bromatologiju dSvijet biljaka d.o.o, Vodnjan, Croatia
e-adresa: miron.sopic@pharmacy.bg.ac.rs
Sažetak
Usled narušavanja ravnoteže između slobodnih radikala i antioksidanasa, dolazi do oksidativnog stresa, patofiziološkog stanja, koje je povezano sa nastankom i razvojem mnogih oboljenja. Kako su se određeni sintetski antioksidansi pokazali kao potencijalno štetni po zdravlje ljudi, sve se više pažnje usmerava ka pronalaženju prirodnih antioksidanasa. Polifenoli, fenolne kiseline, u koje spada i ruzmarinska kiselina, terpeni i terpenoidi su sekundarnii metaboliti biljaka koji poseduju antioksidativne osobine, a mogu se naći u različitim biljnim kompleksima (hidrolatima) i medu. Cilj ovog rada je utvrđivanje in vitro antioksidativne aktivnosti preparata koji predstavlja mešavinu biljnih kompleksa (hidrolata), ruzmarinske kiseline i meda. U "pool" seruma, koji je dobijen mešanjem seruma zdravih pojedinaca, dodavana su serijska razblaženja (100%, 50%, 25%) ispitivanog preparata, kao i serijska razblaženja ispitivanog preparata (100%, 50%, 25%) u prisustvu terc-butil hidroperokisda (TBH), kao prooksidansa. Pored ovih uzoraka, pripremljeni su i uzorci u koje je dodavan troloks (hidrofilni analog vitamina E), kao i troloks u prisustvu TBH. Iz svih uzoraka vršeno je određivanje parametara antioksidativnog stresa: totalnog antioksidativnog statusa (TAS), paraoksonaze-1 (PON1), ukupnog sadržaja sulfhidrilnih grupa (SHG), totalnog oksidativnog statusa (TOS), prooksidativnog-antioksidativnog balansa (PAB) i uznapredovalih produkata oksidacije proteina (AOPP). Takođe, izračunat je i odnos TAS/TOS koji predstavlja kvantitativnu meru odnosa antioksidativnih redukujućih supstanci i oksidujućih supstanci u serumu. Poređenjem vrednosti antioksidativnih parametara TAS, SHG, TAS/ TOS odnosa i parametara ravnoteže prooksidanasa i antioksidanasa PAB za uzorke preparata bez i sa dodatkom TBH, nije uočena značajna razlika, što ukazuje da je i pored dodatka TBH ispitivani preparat održavao svoje antioksidativno delovanje. Ispitivani preparat pokazao je značajno antioksidativno delovanje, što se može pripisati sinergističkom delovanju terpena i terpenoida iz hidrolata, ruzmarinske kiseline, polifenola i ostalih antioksidativnih supstanci iz meda.
Abstract
Disrupted balance between free radicals and antioxidants leads to a pathophysiological state of oxidative stress, which is the underlying cause of many diseases. Since some synthetic antioxidants show some potentially adverse effects, many studies aim to find natural antioxidants. Secondary plant metabolites, such as polyphenols, phenolic acids, which include rosmarinic acid, terpenes and terpenoids have antioxidative properties and can be found in different hydrosols and honey. The aim of this study is to evaluate the in vitro antioxidative activity of the supplement based on hydrosol mixture, rosmarinic acid and honey. A series of diluted (100%, 50%, and 25%) supplements were added to the serum pool, collected from healthy donors. A second series of samples was prepared with the same amounts of supplement and addition of tert-Butyl hydroperoxide (TBH) as a prooxidant. Alongside these samples, sera samples with Trolox (hydrophilic vitamin E analog) and Trolox with TBH were prepared. All these samples were tested for the following oxidative stress parameters: total antioxidative status (TAS), paraoxonase-1 (PON-1), total sulfhydryl groups content (SHG), total oxidative status (TOS), prooxidative-antioxidative balance (PAB) and advanced oxidation protein products (AOPP). TAS/TOS ratio was calculated as a quantitative measurement of antioxidative and oxidative substances ratio in the serum. Antioxidative parameters TAS, SHG, TAS/TOS ratio and prooxidant/antioxidant balance parameter PAB, were not significantly different for the supplemened samples with and without the addition of TBH, which indicates that the supplement keeps its antioxidative properties, despite the addition of TBH. The supplement showed significant antioxidative properties as a result of the synergistic effect of rosmarinic acid, terpenes, and terpenoids from hydrosols and polyphenols and other antioxidant substances from honey.
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