Nishiguchi E, Sindo J, Hamasaki N
Department of Dental Hygiene, Syonan Junior College, Kanagawa, Japan.
Biochim Biophys Acta. 1993 Mar 10;1176(1-2):95-105. doi: 10.1016/0167-4889(93)90183-p.
To clarify the mechanism underlying local anesthetic-induced changes in the shape of human erythrocytes from discocytes to stomatocytes, we treated erythrocytes with lidocaine, a cationic drug. Analysis of the erythrocyte membrane and cytoplasm by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) revealed that the intensities of the stained bands of 62 kDa, 28 kDa and 22 kDa depended on the extent of the shape change induced by lidocaine. The change in the intensity of the 28 kDa band was particularly marked. We identified the cytoplasmic substances, i.e., the 28 kDa and 22 kDa peptides, as carbonic anhydrase (CA) and glutathione peroxidase (GSH Px)1, respectively, by immunoblotting. The 62 kDa peptide was identified as Hb by column chromatography and SDS-PAGE analysis. To identify the protein responsible for the lidocaine-induced shape change, we incorporated CA and GSH Px into ATP-MgCl2-resealed ghosts. The shape of the resealed ghosts changed upon addition of lidocaine, but only in the presence of CA. These results suggest that ATP and CA are required for the shape changes induced by lidocaine.
为了阐明局部麻醉药诱导人红细胞从盘状细胞转变为口形细胞的潜在机制，我们用阳离子药物利多卡因处理红细胞。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）对红细胞膜和细胞质进行分析，结果显示，62 kDa、28 kDa和22 kDa染色带的强度取决于利多卡因诱导的形状变化程度。其中，28 kDa条带强度的变化尤为明显。通过免疫印迹法，我们分别将细胞质物质（即28 kDa和22 kDa肽段）鉴定为碳酸酐酶（CA）和谷胱甘肽过氧化物酶（GSH Px）1。通过柱色谱法和SDS-PAGE分析，将62 kDa肽段鉴定为血红蛋白（Hb）。为了确定导致利多卡因诱导形状变化的蛋白质，我们将CA和GSH Px加入到ATP-MgCl2重封的血影中。加入利多卡因后，重封血影的形状发生了变化，但仅在有CA存在的情况下才会出现这种变化。这些结果表明，ATP和CA是利多卡因诱导形状变化所必需的。
