激光照射对H_2O_2处理的红细胞膜结构影响的AFM分析

Analysis of the structure changes of the laser-irradiated red membrane pre-dealt with H_2O_2 using AFM

本文首次利用原子力显微镜(atomic force microscope,AFM)的超高分辨率功能研究H_2O_2处理引起的红细胞膜超微结构的变化,及低强度He-Ne激光照射H_2O_2处理过的红细胞使膜结构恢复的情况。实验结果表明,无论是H_2O_2作用还是激光照射红细胞整体都能维持双凹圆盘状结构;但用AFM得到的超微结构图显示H_2O_2作用的红细胞膜表面高低起伏明显,面上平均粗糙度增加,出现较宽、较深的孔洞。随着激光照射剂量的增加,红细胞膜表面的高低起伏减弱,面上平均粗糙度下降,孔洞变浅、变窄,细胞表面趋于平滑。分析表明,H_2O_2与红细胞内游离铁离子发生类Fenton反应产生的羟自由基(·OH)损伤红细胞膜的结构,细胞的超微结构发生较明显的变化,细胞膜表面的平均粗糙度明显增大,出现较深、较宽的孔洞;激光照射消除部分自由基的作用,较小剂量的激光照射能使损伤部分减弱,膜表面的平均粗糙度降低,孔洞变浅、变窄;较大剂量的激光照射使膜表面平均粗糙度进一步降低,孔洞进一步变浅、变窄,损伤进一步恢复。

In this article, we using ultra-high resolution of AFM firstly, comparing and analyse erythrocyte mem- brane morphology structure＇s change before and after deal with H2O2, then membranous morphology structure＇s change before and after irradiate by laser. Result： no matter deal with H2O2 or irradiate by laser, the erythroeyte maintain doughnut configuration still, but H2O2 lead to membranous surface have a morphology of peak and valleyappeared by turns. Membranous molecular arrange become to one direction, Surface roughness increased. Laser irra- diate the erythrocyte that be deal with H2O2, the erythrocyte membranous surface not have the clear peak and valley appeared by turns, and membranous molecular arrange become not very clear to one direction. Cell surface tend to smoothness. Surface roughness decreased. Conclusion： H2O2 react with dissociated iron ions that lie in erythrocyte, this reaction produce .OH that＇s harm to erythrocyte membranous structure, this is Fenton-like reaction. This reac- tion lead the cellular ultrastructure occur clear change. Surface roughness increased distinctly. Laser＇s irradiation can eliminate a portion of .OH＇s effect. Small dose of laser irradiation can weak the harm partly. Surface roughness decreased. Big dose of laser irradiation can weak the harm distinctly. Surface roughness decreased clearly.