Journal of Medicinal Materials, 2019, Vol. 24, No. 3 (pp.163 - 168)
DETERMINATION OF HMG-CoA REDUCTASE ENZYMATIC ACTIVITY: A PRACTICAL PROTOCOL FOR SCREENING OF CHOLESTEROL SYNTHESIS INHIBITORS
Nguyen Thuy Duong*, Pham Duc Vinh
Hanoi University of Pharmacy, Vietnam
*Corresponding author: duongnt@hup.edu.vn
(Received May, 14th, 2019)
Summary
Determination of HMG-CoA Reductase Enzymatic Activity: A Practical Protocol
for Screening of Cholesterol Synthesis Inhibitors
The aim of this study is to establish a protocol for determination of HMG-CoA reductase (HMGR) activity by spectrophotometric method based on the consumption rate of NADPH and apply it for evaluation of the HMGR inhibitory effect of selected medicinal plants. The various conditions of enzyme reaction were tested and optimized including enzyme and substrate concentrations, temperature and incubation time duration. The enzyme catalyzed reaction showed a good linearity over time with Km value of 51.3 µM. Furthermore, the IC50 value of atorvastatin (12.31 nM) determined using the present protocol was close to that commonly reported in literature earlier. In addition, among 10 tested plant extracts, 5 exhibited to be potent inhibitors of HMGR with the inhibition of 50% or higher. These findings suggest that this protocol can be used for screening of HMGR inhibitors and further works should be performed to illuminate the effect and mechanism of potential plants detected in the present study.
Keywords: HMG-CoA reductase, Hypercholesterolemia, Inhibitors, Enzyme assay.
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