Serum Alkalen Fosfataz ve Asit Fosfataz Enzimlerinin Aktivileri Üzerine Lizinoprilin In-Vitro Etkisi
Küçük Resim Yok
Tarih
2017
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Hipertansiyon, çoğu yaşlı insanın mustarip olduğu bir hastalıktır. Yüksek tansiyonu olan insanlar genellikle hayatları boyunca sağlıklı bir yaşam sürdürebilmek için kan basıncını düşüren ilaçları kullanmak zorundadırlar. Bu çalışmada, lisiniprol adlı hipertansiyon ilacının kemik yapım ve yıkımının bir göstergesi olan alkalen fosfataz (ALP) ve tartarata dayanıklı asit fosfataz (TrACP) enzimlerinin aktiviteleri üzerine olan etkilerini in vitro olarak incelenmiştir. İnsan serumundan alkalen fosfataz enzimi DEAE-selüloz, asit fosfataz enzimi ise CM-selüloz iyon değişim kromatografisi kullanılarak saflaştırılmıştır. Alkalen ve asit fosfataz enzimlerinin aktiviteleri substrat olarak p-nitrofenilfosfat kullanılarak belirlenmiştir. TrACP enzim aktivitesi bulunurken ortama tartarat ilave edilerek TrACP dışındaki diğer asit fosfataz enzimlerinin inhibe olması sağlanmıştır. Alkalen fosfataz ve tartarata dayanıklı asit fosfataz enzimlerinin aktiviteleri substratın (0,033, 0,1, 0,16, 0,23 ve 0,3 mM) beş farklı konsantrasyonunda inhibitörlü ve inhibitörsüz ortamlarda bulunmuştur. İnhibitörün ise her bir substrat konsantrasyonu için 10-2, 10-3 ve 10-4 M olmak üzere üç farklı derişimi kullanılmıştır. İlacın her iki enzimi de inhibe ettiği tespit edilip, I50 değerleri 0,17 mM ve 0,79 mM olarak hesaplanmıştır. Ayrıca Ki değerleri Lineweaver-Burk grafiğinden yararlanılarak ALP ve TrACP için sırasıylahesaplanmıştır
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 pnitrophenylphosphate 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 I50 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
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 pnitrophenylphosphate 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 I50 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
Açıklama
Anahtar Kelimeler
Kaynak
Celal Bayar Üniversitesi Fen Bilimleri Dergisi
WoS Q Değeri
Scopus Q Değeri
Cilt
13
Sayı
1
