CAJ | 학술논문

采用微孔比色法及荧光分析法,研究抗男性生育化合物棉酚与猪胰腺磷脂酶A2(phospholipaseA2,PLA2,EC3.1.1.4)温育并透析前后对酶活力及荧光的影响.结果表明,棉酚与PLA2不可逆地结合明显地降低了PLA2活力及荧光强度.棉酚对酶活力抑制作用的IC50为35 μmol/L;当其浓度达到80μmol/L时,能够完全抑制PLA2(4.11μmol/L)对合成底物2-硫代十六酰乙基磷酸胆碱(HEPC,0.25 mmol/L)的水解作用.PLA2的最大激发波长与发射波长分别为275 nm,343 nm,荧光强度与酶浓度呈良好的线性关系.棉酚对PLA2的荧光具有较强的淬灭作用.由于PLA2与男性生育密切相关,棉酚对PLA2活力的影响可能是其避孕作用及伴随的副作用的一种新的重要机制.
채용미공비색법급형광분석법,연구항남성생육화합물면분여저이선린지매A2(phospholipaseA2,PLA2,EC3.1.1.4)온육병투석전후대매활력급형광적영향.결과표명,면분여PLA2불가역지결합명현지강저료PLA2활력급형광강도.면분대매활력억제작용적IC50위35 μmol/L;당기농도체도80μmol/L시,능구완전억제PLA2(4.11μmol/L)대합성저물2-류대십륙선을기린산담감(HEPC,0.25 mmol/L)적수해작용.PLA2적최대격발파장여발사파장분별위275 nm,343 nm,형광강도여매농도정량호적선성관계.면분대PLA2적형광구유교강적쉬멸작용.유우PLA2여남성생육밀절상관,면분대PLA2활력적영향가능시기피잉작용급반수적부작용적일충신적중요궤제.
The effect of a male antifertility compound gossypol on the enzymatic activity and fluorescence of porcine pancreatic phospholipase A2 (pp I PLA2, EC 3.1.1.4)has been studied by microplate assay and fluorescence assay on the incubated mixture with or without dialysis. Gossypol interacted with pp I PLA2 resulting in forming a complex and diminishing enzymatic activity and fluorescence intensity. The drug-enzyme interaction was irreversible by aqueous dialysis .The hydrolysis activity of pp I PLA2 (4.11 μmol/L)toward a synthe sized substrate 2-hexadecanoylthio-1-ethyl-phosphorylcholine( HEPC ,0.25 mmol/L)was completely inhibited by 80 μmol/L gossypol with an apparent IC50 of 35 μmol/L. There was a good linearity between fluorescence inten sities and concentrations of pp I PLA2 with maximal excitation and emission wavelength at 275 nm and 343 nm, respectively. Gossypol caused a red shift in the maximal emission of about 5 nm and strongly quenched the fluorescence intensity of pp I PLA2. Since PLA2 is very important to male fertility, the influence of gossypol on PLA2 probably correlates with its mechanism of male antifertility and adverse effect.