丁酸盐不同给药方式诱导K562细胞向红系分化的比较研究

目的研究丁酸盐对血红蛋白合成的诱导作用和不同给药方式对丁酸盐诱导的K562细胞向红系分化的影响。方法以K562细胞为体外模型,采用联苯胺染色定性技术检测细胞内血红蛋白,比较丁酸盐不同浓度、不同给药方式诱导K562细胞前后的阳性细胞率;测量波长为414nm的D(λ)值和醋酸纤维膜电泳检测药物诱导的血红蛋白含量的变化。结果不同浓度的丁酸盐诱导后联苯胺染色阳性细胞率(BZ%)增加4～6倍,细胞平均血红蛋白较用药前增加9~14倍;丁酸盐选择性地刺激HbF合成;单次脉冲式给药BZ%在72 h达高峰,峰值在19%~28%之间,之后迅速下降,约在7~9d接近用药前水平,孵育时间的长短与BZ%上升以及上升后持续时间的长短无关;丁酸盐持续诱导的BZ%变化与一次脉冲式用药总体趋势相似;间断脉冲式用药BZ%在72 h达到峰值后持续保持在20%~30%之间的水平,直至3个周期的用药结束。结论丁酸盐可诱导珠蛋白基因表达,促进血红蛋白的合成,尤其能够选择性地刺激胎儿血红蛋白的合成增加;间断脉冲式用药能够持续诱导K562细胞向红系分化,可作为丁酸盐治疗β-珠蛋白基因缺陷疾病的理想给药方式。

Induction of erythroid differentiation in K562 cells by different butyrate regimens

Objective To investigatethehemoglobinizationinducedby butyrateandobservetheeffectsof differentbutyrate regimenson erythroiddifferentiationof K562cells.Methods K562cells,usedas an in vitro modelsystem,werestainedwith benzidineto assesshemoglobin(Hb)productioninresponseto differenttreatmentregimensof butyrateatvariedconcentrations.Comparisonof thepercentageof benzidine-positivecells(BZ%)in untreatedandbutyrate-treatedK562cellswas performed.Proteinabsorptionat414nm usinga spectrophotometerandcelluloseacetategelelectrophoresiswereemployedto determine thechangesof HbproductioninK562cells.Result TheBZ%increasedby4 to6foldandHbproductionby9to14fold3d afterthecellswereincubatedwithbutyratewhichselectivelypromotedfetalhemoglobin(HbF)productioninK562cells.The BZ%increasedgraduallyandreachedthepeakof19% to28%on day3or4in cellsreceivingpulsetreatmentwith butyrate foronlyonce,followedby a subsequentrapidfallandon day7to9,itdecreasedto thelevelof untreatedK562cells.The lengthof timeforincubationwithbutyratewasnotrelatedto theincrementor themaintenanceof theincreasedlevelof BZ%.Continuoustreatmentwithbutyrateyieldeda similarresultto thatof a singleadministrationof pulsetreatment.In contrast,in cellswithintermittentpulsetreatment,theBZ%reacheda peakafter72h andwas maintained between20%and30%till3cyclesof treatmentwas completed.Conclusion Butyratecan inducethe expressionof globingenesand augmentHb productionespeciallythatof HbF.A sustainederythroiddifferentiationof K562cellscanbe achieved by intermittentpulse treatmentwithbutyratewhichcanbe an idealregimenforchildrenwithbetaglobindiseases.