中文摘要：

目的观察磷脂爬行酶1（phospholipid scramblase 1,PLSCR1）在苦参碱（matrine,MAT）诱导维甲酸（all-trans retinoic acid,ATRA）耐药急性早幼粒细胞白血病（acute promyelocytic leukemia,APL）细胞分化的表达,并探讨其与环磷酸腺苷（cyclic adenosine monophosphate,cAMP）/蛋白激酶A（protein kinase A,PKA）途径的相关性。方法以ATRA敏感的APL细胞系NB4以及ATRA耐药株NB4-R1为研究对象。通过NBT还原实验以及流式细胞仪（CD11 b）检测0.1 mmol/L MAT联合1μmol/L ATRA对两细胞株分化标记的影响;采用Western blot、实时荧光定量PCR技术测定细胞PML/RARα、PLSCR1蛋白及基因表达;同时应用PKA拮抗剂H89联合作用,观察细胞分化抗原及上述蛋白/基因表达的变化。结果 MAT联用ATRA能明显提高NB4-R1细胞NBT及CD1 1 b阳性率,并显著下调PML/RARα融合蛋白/基因的表达（P〈0.05,P〈0.01）。单用ATRA能使NB4细胞PLSCR1在蛋白及mRNA水平表达均得到明显增强（P〈0.01）,而在NB4-R1细胞内同样表达上调,但仅蛋白水平差异有统计学意义（P〈0.01）。在联用MAT后,两细胞株PLSCR1蛋白表达进一步上升（P〈0.01）,并且NB4-R1细胞mRNA表达水平差异有有统计学意义（P〈0.05）。上述作用均能被10μmol/L H89逆转（P〈0.05,P〈0.01）。结论 MAT联合ATRA能显著诱导NB4-R1细胞获得再分化,同时抑制PML/RARα融合基因/蛋白的表达,这可能与其上调PLSCR1表达有关。

英文摘要：

Objective To observe the expression of phospholipid scramblase 1（PLSCR1） in matrine（MAT） induced differentiation of all-trans retinoic acid（ATRA） resistant acute promyelocytic leukemia（APL） cells,and to explore its correlation to cyclic adenosine monophosphate（cAMP）/protein kinase A（PKA） signal pathway.Methods NB4（an APL cell line sensitive to ATRA） and NB4-R1（a resistant strain of ATRA） were observed as subjects in this study.Effects of combined treatment of 0.1mmol/L MAT and 1 μmol/L ATRA on the differentiation of two cell lines were detected using nitroblue tetrazolium（NBT） reduction test and flow cytometry（CD11b）.Expressions of PML/RARa and PLSCR1 protein/gene were detected using Western blot and Real-time fluorescence quantitative PCR assay.Meanwhile,H89,PKA antagonist,was used to observe cell differentiation antigen and changes of aforesaid proteins and genes.Results MAT combined ATRA could significantly elevate positive rates of NBT and CD11 b in NB4-R1 cells,and significantly down-regulate the expression of PML/RARa fusion protein/gene（P 0.05,P 0.01）.ATRA used alone could obviously enhance the expression of PLSCR1 in NB4 cells at protein and mRNA levels（P 0.01）.But the expression of PLSCR1 was up-regulated in NB4-R1 cells,but with statistical difference only at the protein level（P 0.01）.In combination of MAT,PLSCR1 protein expression was further elevated in the two cell lines（P0.01）.Besides,there was statistical difference in mRNA expressions in NB4-R1 cells（P0.05）.All these actions could be reversed by treatment of 10μmol/L H89（P 0.05,P 0.01）.Conclusion MAT combined ATRA could significantly induce the differentiation of NB4-R1 cells,and inhibit the expression of PML/RARα fusion gene/protein,which might be associated with up-regulating PLSCR1 expression.