메나디온에 의한 혈소판 내 칼슘 변화측정시 형광 색소 사용의 문제점

• 영문명: Infeasibility of Measuring Ca2+ in Menadione-Exposed Platelets Using Fluorescent Dyes
• 발행기관: 대한약학회
• 저자명: 정선화(Sun Hwa Chung) 이무열(Moo Yeol Lee) 이주영(Joo Young Lee) 정승민(Seung Min Chung) 정진호(Jin Ho Chung)
• 간행물 정보: 『약학회지』제41권 제6호 (1997년), 749~755쪽, 전체 7쪽
• 주제분류: 의약학 > 기타의약학
• 파일형태: PDF
• 발행일자: 1997.12.31

영문 초록

It has been reported that dose-dependent Ca2+ increase by menadione in platelets could be measured by fluorescent dye, quin-2. The problems will be described here relating to measuring Ca2+ in menadione-exposed platelets using fura-2 and fluo-3, widely used fluorescent indicators. Additions of menadione to fura-2 loaded platelets and their lysates resulted in marked reduction in fluorescence intensity at both 340nm (Ca2+-unbound form) excitation wavelengths. Fura-2 excitation spectra were overlapped with UV-visible absorption spectra of menadione, suggesting that light absorption by menadione itself could quench fluorescence generated by fura-2. Next approach was to use fluo-3 which has the higher wavelength (490nm) of excitation. Previous work demonstrated that treatment with probenecid to platelets was required to prevent fluo-3 dye leakage. However, probenecid itself was proven to be inadequate to measure the concentration of intracellular Ca2+ by reducing menadione-induced cytotoxicity in platelets. Our results suggest that it is not feasible to measure Ca2+ in platelets by using fura-2 and fluo-3 in the presence of probenecid, and cautions should be taken to measure changes of intracellular Ca2+ levels by fluorescent dyes following chemical exposure.

키워드

Menadione Platelets Intracellular Ca2+ Cytotoxicity Fura-2 Fluo-3 Probenecid.