In Vitro Inhibition of Human Plasma and Erythrocyte AChE by Various Drugs

Abstract

In the present study, the in vitro effects of eight structurally different antibiotics cefazolin, kanamycin, gentamicin, clindamycin, ivermectin and ampicillin and painkiller metamizole and diclofenac on human erythrocyte and plasma acetylcholinesterase activity AChE; EC 3.1.1.7 were investigated. For this, IC50 and KI values of the drugs were calculated for determining the inhibition caused by these drugs. Acetylcholinesterase activity was determined spectrophotometrically, according to Ellman’s method. Then the enzyme activity exposed to the drugs with different concentrations was measured. AChE activity vs. drug concentration and Mihealis-Menten graphs were obtained. Lastly, IC50 and KI values were estimated from these graphs. IC50 values were estimated to be 1.78 x10-8 M of diclofenac for plasma AChE and 0.85x10-8 M of gentamicin for erythrocyte AChE. K values were 1.36 x10-8 M for diclofenac and 1.16 x10-8 M for gentamicin. Also, IC50 and KI values for ivermectin, clindamycin, kanamycin, ampicillin were obtained. Diclofenac exhibited the highest inhibitory effect on plasma AChE activity. On the other hand, gentamicin showed the highest inhibitory effect on erythrocyte AChE activity, which seems to be better for inhibition of AChE than those reported previously.

Keywords

• Acetylcholinesterase,enzyme inhibition,enzyme assay,medicinal drug,human erythrocyte and plasma,in vitro kinetics

İnsan Plazma ve Eritrosit AChE’nin Çeşitli İlaçlarla in Vitro İnhibisyonu

Abstract

B u çalışmada; yapısal olarak farklı olan sekiz antibiyotik sefazolin, kanamisin, gentamisin, klindamisin, ivermektin ve ampisilin ve ağrı kesicinin metamizol ve diklofenak insan eritrosit ve plazma asetilkolinesteraz AChE; EC 3.1.1.7 enzimi üzerine in vitro etkileri araştırılmıştır. Bu amaçla; farklı derişimlere sahip ilaçların neden olduğu inhibisyonu belirlemek için ilaçların IC50 ve KI değerleri hesaplandı. Asetilkolinesteraz aktivitesi spektrofotometrik olarak Ellman yöntemiyle belirlendi. Daha sonra, ilaçlara maruz kalan enzim aktivitesi ölçüldü. AChE aktivitesine karşı ilaç derişimi ve Michealis-Menten grafikleri çizildi. Son olarak, bu grafikler yardımıyla IC ve K değerleri hesaplandı. IC değerleri, plazma AChE’sine diklofenak için 1.78 x10-8 M ve eritrosit AChE’sine gentamisin için 0.85 x10-8 M olduğu belirlendi. KI değerleri ise diklofenak için 1.36 x10-8 M ve gentamisin için 1.16 x10-8 M olarak hesaplandı. Ayrıca, ivermektin, klindamisin, kanamisin, ampisilin için ICve KI değerleri elde edildi. Diklofenak, plazma AChE aktivitesi üzerine gentamisin de eritrosit AChE aktivitesi üzerine en yüksek inhibitör etkisi göstermiştir. Bulunan bu değerlerin daha önce yapılan benzer çalışmalardaki inhibisyon değerlerinden daha yüksek olduğu görülmektedir [1,2]

Keywords

• Asetilkolinesteraz,enzim inhibisyonu,enzim analizi,tıbbi ilaçlar,insan eritrositi ve plazması,in vitro kinetikleri

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