Normal Tension Glaucoma Clinical Trial
Official title:
Evaluation of Systemic Glutathione Level in Patients With Normal Tension Glaucoma
The purpose of this study is to evaluate whether systemic glutathione level is decreased in patients with normal tension glaucoma.
Normal tension glaucoma is one of the most common cause of primary open angle glaucoma in
Korea. Even though, we still do not know what the cause is. Only IOP-lowering drugs are the
currently available therapeutic method. Glutathione is one of the mostly high concentrated
intracellular antioxidants. We find apoptosis of neuronal cell in the mouse retina by
systemic depletion of glutathione using buthionine sulfoximine. Glutathione is also known to
be reduced in primary open angle glaucoma with high IOP. So we are planning to evaluate
systemic glutathione level in normal tension glaucoma.
Blood samples will be obtained by venipuncture to the antecubital vein of normal tension
glaucoma patients and age-matched normal controls. Five milliliters of blood will be
collected in EDTA-treated tubes (to prevent oxidation). Thirty microliters of blood will be
then transferred into centrifuge tubes, and will be added 33.3 microliter of
5-sulfosalicylic acid (SSA), 100mg/mL within 10 minutes from the blood collection.Each
sample will be then diluted with 936.7 microliter sodium phosphate buffer (pH 7.5), and the
content of each tube will be mixed rapidly in a centrifuge at 13,000 rpm for 5 minutes. A
portion of supernatant (150 microliter) will be then collected into clean centrifuge tubes
and immediately cooled at -70°C. To each well of a 96-well plate, 150 microliter of daily
buffer (3 mg NADPH into 10 mL sodium phosphate buffer), 50microliter of DTNB solution, and
25 microliter of standards or samples will be added in quadruplicate, and the plate will be
incubated at 37 °C for 3 minutes.Finally, 25 microliter GSSGR will be added to the previous
mixture, and the plate will be read at 410 nm using a 96-well plate reader(UV
spectrophotometer).
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Observational Model: Case Control, Time Perspective: Cross-Sectional
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