Publication: Hemodiyaliz hastalarında bazı antibiyotiklerin polimorf nüveli lökosit fonksiyonları ile oksidatif stres ve antioksidan enzimler üzerine etkisinin in vitro araştırılması
Abstract
1. ÖZET Çalışmamızda, terapötik konsantrasyonlarda vankomisin (40 µg/ ml), daptomisin (60 µg/ ml), linezolid (19 µg/ ml), levofloksasin (7 µg/ ml), fusidik asit (36 µg/ ml) ve meropenemin (24 µg/ ml) hemodiyaliz hastalarının polimorf nüveli lökosit (PNL) fonksiyonları (fagositik aktivite ve hücre içi öldürme aktivitesi), malondialdehit (MDA) düzeyleri, glutatyon peroksidaz (GSH-Px) ve miyeloperoksidaz (MPO) aktiviteleri üzerine olan etkileri in vitro araştırılmıştır. PNL’ler venöz kandan ficoll-hypaque gradient santrifüj yöntemi ile izole edilmiştir. Fagositik aktivite ve hücre içi öldürme aktivitesi tayini Alexander’ın yöntemi modifiye edilerek yapılmıştır. Lipid peroksidasyonun göstergesi olarak MDA düzeyi Beuge’nin yöntemi, MPO aktivitesio-dianisidin yöntemi modifiye edilerek, GSH-Px tayini, Cayman’ın GSH-Px kiti kullanılarak yapılmıştır. Tüm sonuçlar istatistiksel olarak değerlendirilmiştir. Antibiyotik içermeyen kontrol deneyde, HD hastalarının HD sonrası PNL’lerinin fagositik aktivitesinin HD öncesine göre azaldığı (p<0.05), hücre içi öldürme aktivitesinin ise değişmediği saptanmıştır (p>0.05). Terapötik konsantrasyonlarda vankomisin, daptomisin, linezolid, levofloksasin, fusidik asit ve meropenemin HD hastalarının HD öncesi ve HD sonrası PNL’lerinin her iki fonksiyonunu etkilemediği saptanmıştır (p>0.05). Terapötik konsantrasyonlarda vankomisin, levofloksasin, fusidik asit ve meropenemin HD hastalarının HD öncesi PNL’lerinin MDA düzeyini artırdığı, HD sonrası ise MDA düzeyini azalttığı saptanmıştır (p<0.05). Daptomisin ve linezolidin ise HD hastalarının HD öncesi ve HD sonrası PNL’lerinin MDA düzeyini değiştirmediği bulunmuştur (p>0.05). Terapötik konsantrasyonlarda vankomisin, daptomisin, linezolid, levofloksasin, fusidik asit ve meropenemin HD hastalarının HD öncesi ve HD sonrası PNL’lerinin GSH-Px ve MPO aktivitesini etkilemediği saptanmıştır (p>0.05). Anahtar Sözcükler: Glutatyon peroksidaz, hemodiyaliz, miyeloperoksidaz, oksidatif stres, polimorf nüveli lökosit fonksiyonları. 2.
In Vitro Investigation of The Effects of Some Antibiotics on Polymorphonuclear Leukocyte Functions, Oxidative Stress and Antioxidant Enzymes in Hemodialysis Patients. In our study were investigated the in vitro effects of vancomycin (40 µg/ ml), daptomycin (60 µg/ ml), linezolid (19 µg/ ml), levofloxacin (7 µg/ ml), fusidic acid (36 µg/ ml) and meropenem (24 µg/ ml) at therapeutic concentrations and malondialdehyde (MDA) levels, glutathione peroxidase (GPx) and myeloperoxidase (MPO) activities on polymorphonuclear leukocyte (PMN) functions (phagocytic and intracellular killing activity) of hemodialysis patients. PMNs were isolated from venous blood by ficoll-hypaque gradient centrifugation method. Phagocytic activity and intracellular killing activity were assayed by modified Alexander’s method. MDA level was detected by Beuge method which is as an indicator of lipid peroxidation. GPx activity was determined by using Cayman’s GPx kit. PMN’s MPO activity was determined by modified o-dianisidine method. All results were evaluated by statistical methods. In the control experiments with antibiotic-free which was found that PMN’s phagocytic activity of HD patients decreased after HD compared to before HD (p<0.05), although the intracellular killing activity did not change (p>0.05). It was detected that vancomycin, daptomycin, linezolid, levofloxacin, fusidic acid and meropenem at therapeutic concentrations did not affect both PMN functions of HD patients before and after HD (p>0.05). It was observed that vancomycin, levofloxacin, fusidic acid and meropenem at therapeutic concentrations increased PMN’s MDA levels of HD patients before HD but decreased MDA levels after HD (p<0.05). Daptomycin and linezolid did not change PMN’s MDA levels of HD patients before and after HD (p>0.05). It was observed that vancomycin, daptomycin, linezolid, levofloxacin, fusidic acid and meropenem at therapeutic concentrations did not changed PMN’s GPx and MPO activity of HD patients before and after HD (p>0.05). Glutathione peroxidase, hemodialysis, myeloperoxidase, oxidative stress, polymorphonuclear leukocyte functions.
In Vitro Investigation of The Effects of Some Antibiotics on Polymorphonuclear Leukocyte Functions, Oxidative Stress and Antioxidant Enzymes in Hemodialysis Patients. In our study were investigated the in vitro effects of vancomycin (40 µg/ ml), daptomycin (60 µg/ ml), linezolid (19 µg/ ml), levofloxacin (7 µg/ ml), fusidic acid (36 µg/ ml) and meropenem (24 µg/ ml) at therapeutic concentrations and malondialdehyde (MDA) levels, glutathione peroxidase (GPx) and myeloperoxidase (MPO) activities on polymorphonuclear leukocyte (PMN) functions (phagocytic and intracellular killing activity) of hemodialysis patients. PMNs were isolated from venous blood by ficoll-hypaque gradient centrifugation method. Phagocytic activity and intracellular killing activity were assayed by modified Alexander’s method. MDA level was detected by Beuge method which is as an indicator of lipid peroxidation. GPx activity was determined by using Cayman’s GPx kit. PMN’s MPO activity was determined by modified o-dianisidine method. All results were evaluated by statistical methods. In the control experiments with antibiotic-free which was found that PMN’s phagocytic activity of HD patients decreased after HD compared to before HD (p<0.05), although the intracellular killing activity did not change (p>0.05). It was detected that vancomycin, daptomycin, linezolid, levofloxacin, fusidic acid and meropenem at therapeutic concentrations did not affect both PMN functions of HD patients before and after HD (p>0.05). It was observed that vancomycin, levofloxacin, fusidic acid and meropenem at therapeutic concentrations increased PMN’s MDA levels of HD patients before HD but decreased MDA levels after HD (p<0.05). Daptomycin and linezolid did not change PMN’s MDA levels of HD patients before and after HD (p>0.05). It was observed that vancomycin, daptomycin, linezolid, levofloxacin, fusidic acid and meropenem at therapeutic concentrations did not changed PMN’s GPx and MPO activity of HD patients before and after HD (p>0.05). Glutathione peroxidase, hemodialysis, myeloperoxidase, oxidative stress, polymorphonuclear leukocyte functions.