补骨脂素加长波紫外线的光化学疗法诱导NB4细胞凋亡及对Caspase-8、3蛋白表达的影响  

Photochemotherapy with Psoralen and Ultraviolet A Induced Apoptosis of NB4 Cells and Its Effects on Caspase- 8 and Caspase- 8 Protein Expressions

作　　者：孙淑君[1,2] 赵伟杰[3] 向阳[2] 陈楠楠[2] 孙锋[2] 常晓慧[2] 成玉斌[2] 黄世林[2] 

机构地区：[1]解放军总医院军医进修学院 [2]解放军第210医院中医血液科,辽宁大连116021 [3]大连理工大学精细化工国家重点实验室,辽宁大连116012

出　　处：《中国中西医结合杂志》2013年第4期502-505,共4页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.30472257)

摘　　要：目的研究补骨脂素(psoralen,PSO)加长波紫外线(ultravioletA,UVA)的光化学疗法(PUVA)对人白血病NB4细胞凋亡和细胞凋亡信号通路的影响。方法体外培养NB4细胞,利用流式细胞术检测不同浓度的PSO(0、5、10、20、40μL)、在不同照射时间(0、5min)、波长为360nm的UVA干预后的细胞凋亡率;利用透射电镜检测细胞超微结构的改变;采用免疫细胞化学法(ICC)检测凋亡信号通路中半胱氨酸天冬氨酸蛋白酶-8(Caspase-8)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达情况。结果不同浓度的PSO经0、5min UVA照射后,NB4细胞的凋亡率呈剂量与时间依赖性的增加,而于40μg/mL浓度的PSO联合5min UVA照射时达峰值,且二者呈现交互作用(P<0.01)。PUVA处理后的NB4细胞超微结构出现明显的凋亡形态学的改变。PSO、UVA及PUVA均上调Caspase-8、Caspase-3蛋白的表达,但PUVA上调Caspase-3的作用于12h显著强于前两者(P<0.01),呈时间依赖性,PSO浓度与时间呈现交互作用(P<0.01)。结论 PU-VA的最佳方案组合为40μg/mL浓度的PSO联合5min UVA照射,PUVA可体外激活Caspase-8、Caspase-3基因,具有诱导NB4细胞凋亡作用。Objective To study the regulatory effects of psoralen （PSO） plus ultraviolet A （UVA）, which is PUVA, on cell apoptosis of human leukemia cell line NB4 and signal pathway of cell apoptosis. Methods Human leukemia cell line NB4 was cultured in vitro. The NB4 cells were treated with PSO extracted from Chinese medicine psoralea fruits at different concentrations （0, 5, 10, 20 and 40 μL） plus UVA of wave length 360 nm at different irradiation time points （0 and 5 min）. The apoptosis ratio was detected by flow cytometry （FCM）. The ultrastructure changes were observed using transmission electron microscope （TEM）. The expressions of Caspase-8 and Caspase-8 protein were detected by immunocytochemical method （ICC）. Results After treatment of PSO at different concentrations with a 0 and 5-min exposure of UVA, the apoptosis rate of NB4 cells increased dose-and time-dependently, and was up to peak after treatment of PSO at 40 μg/mL with 5-min exposure of UVA. An interaction was shown between the two factors （P0.01）. There were obvious morphological apoptosis of NB4 cells under TEM after treated with PUVA. The expressions of Caspase-3 and Caspase-8 protein were up-regulated by PSO, UVA, and PUVA, but the effects of PUVA on Caspase-3 protein were stronger than PSO and UVA at 12 h time-dependently （P0.01）. An interaction was shown between the concentration of PSO and time of UVA （P0.01）. Conclusions The optimal combination of PUVA was PSO in 40 μg/mL and 5-min exposure of UVA. PUVA could induce the apoptosis of NB4 cells and in vitro activate Caspase-3 and Caspase-8 genes.

关 键 词：补骨脂素 长波紫外线 NB4细胞 凋亡 半胱氨酸天冬氨酸蛋白酶-8(Caspase-8) 半胱氨酸天冬氨酸蛋白酶-3(Caspase-3) 

分 类 号：R733.7[医药卫生—肿瘤]