鼠尾草酸增加1,25(OH)_2D_3诱导HL-60细胞分化及其机制研究  

Enhancement of differentiation induction of HL-60 cells by 1,25-dihydroxyvitamin D_3 in combination with carnosic acid

作　　者：任立红[1] 陈娟娟[1] 安慧萍[1] 

机构地区：[1]哈尔滨医科大学第二临床学院儿科,黑龙江哈尔滨150086

出　　处：《中国当代儿科杂志》2008年第1期55-59,共5页Chinese Journal of Contemporary Pediatrics

摘　　要：目的已证实1,25(OH)2D3可以诱导HL-60细胞向成熟单核细胞分化,但其不足之处是高钙血症和形成耐药株。该实验应用鼠尾草酸(CA)联合1,25(OH)2D3研究对HL-60细胞分化的影响及引起细胞内活性氧(ROS)水平和细胞内钙离子浓度的改变。方法应用鼠尾草酸,1,25(OH)2D3单独和联合应用处理HL-60细胞,共分为5组:空白对照组;低浓度1,25(OH)2D3(1nmol/L)组、鼠尾草酸组;联合处理组(10μmol/L鼠尾草酸和1nmol/L1,25(OH)2D3);高浓度1,25(OH)2D3(100nmol/L)组。每24h监测1次,通过四唑氮蓝(MTT)法观察细胞生长,共72h;收集处理后72h的HL-60细胞,通过光学显微镜观察细胞形态,用流式细胞仪(FCM)检测不同处理组细胞周期及单核细胞分化标记CD14的表达,ROS和细胞内钙离子浓度。结果72h后,联合处理组HL-60细胞与空白对照组相比,细胞增殖明显受抑制(Ab=0.560±0.020;P<0.01),细胞形态具有成熟单核细胞特征,CD14的表达率升高(57.62%;P<0.01),G0/G1期细胞显著增多,ROS水平下降[(52.67±10.76)%,P<0.01];联合处理组细胞内钙离子水平同空白对照组比较差异无显著性(115.64±17.74nmol/L,P>0.05),但与高浓度1,25(OH)2D3组相比细胞内钙离子水平明显下降(P<0.01)。结论鼠尾草酸可增强1,25(OH)2D3对HL-60细胞的诱导分化、增强抑制HL-60细胞增殖作用,细胞阻滞于G0/G1期,细胞内活性氧水平降低,这可能与细胞的诱导分化机制有关,且这一联合作用不增高细胞内钙离子水平。Objective 1, 25-dihydroxyvitamin D3 [ 1, 25 （OH）2 D3 ] is a potent inducer of differentiation in myeloid leukemia cells, but its clinical use is limited due to its hyperealcaemic effect and resistance. Carnosic acid is a plantderived polyphenol food preservative with ehemoprotective effects against eareinagens. Recent research has shown that carnosie acid potentiates the effects of 1,25 （OH） 2 D3 on differentiation of human leukemia cells. This study examined the effects of 1, 25 （OH）2D3 in combination with earnosie acid on monoeytie differentiation as well as intracellular reactive oxygen species （ROS） and Ca^2＋ levels in human leukemia HL-60 cells. Methods HL-60 cells were randomly treated with 1 nmol/L 1,25 （OH）2D3, 100 nmol/L 1,25 （OH）2D3, 10 μmol/L carnosic acid, a combination of 1 nmol/L 1,25 （OH）2D3 and 10 μmol/L carnosic acid or placebo. Cell growth was observed by MTF assay for 72 hrs at an interval of 24 hrs. Cells were harvested after 72 hrs of culture. Morpbologic features of the cells were observed by microscopy. Flow eytometry was used to detect cell cycle, monoeytie differentiation marker CD14 expression, and intracellular ROS and Ca^2＋ levels. Results A combination of 1 nmol/L 1,25 （OH）2 D3 and 10 μmol/L earnosic acid resulted in greater proliferation inhibition （ Ab： 0.560± 0.020 vs 1. 482 ± 0. 327 ; P 〈 0.01 ）, mature monoeytic features, GO/G1 cell arrest, higher CD14 expression （57.62 ± 0.817% vs 2.76±0.828% ; P 〈0.01 ）, lower intracellular ROS levels （52.67 ± 10.76% vs 86.46 ±40. 52% ; P〈0.01 ） and similar intracellular Ca^2＋ levels in HL-60 cells when compared with the placebo group. The ability of a combination of 1 nmol/L 1,25 （OH）2D3 and 10 μmol/L carnosic acid to inhibit the proliferation and induce the differentiation of HL-60 cells was similar to that of 100 nmol/L 1,25 （OH） 2 D3, while the intracellular Ca^2＋ level （115.64 ±17.74 nmol/L vs 185.75 ± 27.38 nmol/L） was significantly lower than that in the

关 键 词：1 25(OH)2D3 鼠尾草酸 细胞分化 活性氧 HL-60细胞 

分 类 号：R285[医药卫生—中药学]