【摘要】 目的:从白眉蝮蛇蛇毒分离纯化L-氨基酸氧化酶,并研究其对肺组织的影响.方法:通过Heparin-Sepharose FF及Q-Sepharose HP从白眉蝮蛇蛇毒中分离纯化得到一个L-氨基酸氧化酶,命名为ABU-LAO.给Balb/c小鼠静脉注射50,5,0.5μg药物或生理盐水100μL,每组6只,6 h或24 h后取材,肺组织常规切片,HE染色.高倍镜视野下计算平均红细胞数(RBCs)、平均肺泡内多形核白细胞数(IAPLs)、平均肺泡隔多形核白细胞数(ASPLs);用图像分析仪测平均肺泡面积(MACA).结果:纯化ABU-LAO并在SDS-PAGE上测定其表观M r约为60000.与生理盐水组相比RBCs,IAPLs,ASPLs,MACA在注射ABU-LAO后有显著改变(P<0.0125).结论:通过两步层析法分离纯化得到ABU-LAO,它能诱导小鼠发生严重的急性肺损伤.更多还原

【Abstract】 AIM: To purify the L-amino acid oxidase from Agkistrodon blomhoffii ussurensis venom and study its effect on lung.METHODS: An L-amino acid oxidase was purified from the snake venom of Agkistrodon blomhoffii ussurensis(ABU-LAO) by using a combination of Heparin-Sepharose FF column and Q-Sepharose HP column.The BALB/c mice were injected iv with 50,5,0.5 μg ABU-LAO or saline. At 6 or 24 h following injection,mice were sacrificed and the lungs were dissected.Sections(4 μm) were prepared and stained with hematoxylin-eosin for light microscopic observation.The mean numbers of red blood cells(RBCs),intra-alveolar polymorphonuclear leucocytes(IAPLs),alveolar septa polymorphonuclear leucocytes (ASPLs) in a highpower field and mean alveolar cavity area(MACA) with image software were calculated.RESULTS: ABU-LAO was purified to electrophoretic homogeneity.The molecular weight of the enzyme is 60 kD when examined by SDS-polyacrylamide gel electrophoresis.The calculations of RBCs,IAPLs,ASPLs and MACA in mice treated with ABU-LAO were significantly higher than those treated with saline(P<0.0125).CONCLUSION: ABU-LAO was(purified) by a two-step chromatographic process.The enzyme can induce serious acute lung injury when being injected into mice.更多还原