Serum Alkalen Fosfataz ve Asit Fosfataz Enzimlerinin Aktivileri Üzerine Lizinoprilin İn Vitro Etkisi / The in vitro effect of lisonopril on serum alkaline phosphatase and acid phosphatase enzymes activity

Hipertansiyon, cogu yasli insanin mustarip oldugu bir hastaliktir. Yuksek tansiyonu olan insanlar genellikle hayatlari boyunca saglikli bir yasam surdurebilmek icin kan basincini dusuren ilaclari kullanmak zorundadirlar. Bu calismada, lisiniprol adli hipertansiyon ilacinin kemik yapim ve yikiminin bir gostergesi olan alkalen fosfataz (ALP) ve tartarata dayanikli asit fosfataz (TrACP) enzimlerinin aktiviteleri uzerine olan etkilerini in vitro olarak incelenmistir. Insan serumundan alkalen fosfataz enzimi DEAE-seluloz, asit fosfataz enzimi ise CM-seluloz iyon degisim kromatografisi kullanilarak saflastirilmistir. Alkalen ve asit fosfataz enzimlerinin aktiviteleri substrat olarak p-nitrofenilfosfat kullanilarak belirlenmistir. TrACP enzim aktivitesi bulunurken ortama tartarat ilave edilerek TrACP disindaki diger asit fosfataz enzimlerinin inhibe olmasi saglanmistir. Alkalen fosfataz ve tartarata dayanikli asit fosfataz enzimlerinin aktiviteleri substratin (0,033, 0,1, 0,16, 0,23 ve 0,3 mM) bes farkli konsantrasyonunda inhibitorlu ve inhibitorsuz ortamlarda bulunmustur. Inhibitorun ise her bir substrat konsantrasyonu icin 10 -2 , 10 -3 ve 10 -4 M olmak uzere uc farkli derisimi kullanilmistir. Ilacin her iki enzimi de inhibe ettigi tespit edilip, I 50 degerleri 0,17 mM  ve 0,79 mM olarak hesaplanmistir. Ayrica K i degerleri Lineweaver-Burk grafiginden yararlanilarak ALP ve TrACP icin sirasiyla 6,98x10 -4 , 7,06x10 -4 olarak hesaplanmistir. Hypertension is a disease that most elderly people suffer. People with high blood pressure often obliged to use the drug that lowers blood pressure in order to maintain a healthy lifestyle throughout their lives. In this study, the in vitro effect of lisonopril on serum alkaline phosphatase and acid phosphatase enzymes which are essential for normal bone formation and mineralization activities was investigated. Alkaline phosphatase enzyme was purified using DEAE-cellulose, acid phosphatase enzyme was purified using CM-cellulose ion exchange chromatography from human serum. The activities of alkaline and acid phosphatase were determined using p-nitrophenylphosphate as the substrate. When finding out the activity of TrACP enzyme, adding tartrate to the environment provided inhibition of other acid phosphatase enzymes than TrACP. The activities of alkaline phosphatase and acid phosphatase, which is resistant to tartrate, have been found at five different concentrations of substrates (0.033, 0.1, 0.16, 0.23 and 0.3 mm) in environment with inhibitor and without inhibitor. While three different concentrations of the inhibitor, including 10 -2 , 10 -3 and 10 -4 , was used for each substrate concentration. It was found that the drug inhibit both enzymes, and I 50 value was calculated as 0.17 mm and 0.79 mm. Furthermore, Ki values were calculated as 6.98x10 -4 , 7.06x10 -4 for ALP and TrACP respectively by utilizing the Lineweaver-Burk plot.