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目的探讨两性霉素B作用下急性白血病骨髓基质细胞上连接蛋白43(Cx43)表达及对细胞间隙连接通讯(GJIC)功能的影响。方法体外Dexter法培养13份急性白血病骨髓基质细胞(2ml/份),传代后加入两性霉素B(5×10-6mol/L)(1ml/份);采用流式细胞术及激光共聚焦显微镜检测加药前后Cx43蛋白表达的变化,采用荧光漂白恢复技术(FRAP)比较加药前后GJIC功能的差异。结果加药后急性白血病骨髓基质细胞上Cx43的表达的阳性率为(35.9±4.06)%,与加药前(38.75±3.95)%相比无明显变化但出现定位异常;加药前基质细胞的荧光强度在淬灭后缓慢恢复,而加药后基质细胞的荧光在淬灭后400s仍未恢复,GJIC功能被阻断。结论两性霉素B可影响Cx43在急性白血病骨髓基质细胞上的定位并阻断其GJIC功能。
Objective To investigate the effect of amphotericin B on the expression of connexin 43 (Cx43) and the function of GJIC in acute leukemia bone marrow stromal cells. Methods Thirteen acute leukemia bone marrow stromal cells (2ml / part) were cultured by Dexter method in vitro. After subculture, amphotericin B (5 × 10-6mol / L) was added (1ml / dose). Flow cytometry and confocal laser scanning microscopy The changes of Cx43 protein expression before and after dosing were detected. Fluorescence bleach recovery (FRAP) was used to compare the function of GJIC before and after dosing. Results The positive rate of Cx43 on acute leukemia bone marrow stromal cells was (35.9 ± 4.06)%, which was not significantly changed compared with that before administration (38.75 ± 3.95)%, but there was abnormal localization. Fluorescence intensity slowly recovered after quenching, while the fluorescence of stromal cells did not recover after 400 d after quenching, and GJIC function was blocked. Conclusion Amphotericin B can affect the localization of Cx43 on acute leukemia bone marrow stromal cells and block its GJIC function.