两性霉素B对急性白血病骨髓基质细胞上Cx43表达及GJIC功能的影响

Effects of Amphotericin B on Connexin43 expressions and interacellular communicating function of acute leukemia bone marrow stromal cells

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 observe the effects of Amphotericin B （AB） on Connexin43 expressions and interaceUular communicating function of acute leukemia bone marrow stromal cells （ ABMSCs ）. Methods Bone marrow stromal cells from acute leukemics were cultured by Dexter in vitro, and AB（ 5 × 10^-6mol/L） was added after the first passage, and the Cx43 expression levels were detected through flow cytometry technique and eonfocal laser scanning microscopy. The differences of gap junction intercellular communication （GJIC）were examined by using fluorescence redistribution after photobleaching technique. Results Cx43 expression on ABMSCs after adding AB was （35.9 ± 4. 06）%, which showed no significant differences from the expression before adding AB, which was （38.75± 3.95% ）, while abnormal position of Cx43 was observed after adding AB. Fluorescence intensity of coal maceral of ABMSCs before adding AB recovered slowly after photobleaehing, but after adding AB, no recovery in ABMSCs was observed. GJIC function was blocked. Conclusion AB can block GJIC function of ABMSCs.