苦参碱联合顺铂对SH-SY5Y细胞药物敏感性及TrkB表达的影响

目的探讨不同浓度苦参碱对神经母细胞瘤SH-SY5Y细胞存活及凋亡的影响,及苦参碱降低顺铂化疗耐药性的作用及可能机制。方法分别以全反式维甲酸(ATRA)、脑源性神经生长因子(BDNF)、顺铂、苦参碱处理SH-SY5Y细胞,实验分对照组、ATRA诱导的顺铂干预组、顺铂干预组、苦参碱干预组、苦参碱联合顺铂干预组。应用四甲基偶氮蓝比色法检测苦参碱与顺铂对SH-SY5Y细胞存活率的影响,流式细胞仪检测苦参碱与顺铂对SH-SY5Y细胞凋亡率的影响,Western-blot方法检测SH-SY5Y细胞的p-TrkB表达。结果除ATRA诱导的顺铂干预组外,其余各组SH-SY5Y细胞的存活率及凋亡率与对照组比较,差异均有统计学意义(P<0.05);顺铂干预组与ATRA诱导的顺铂干预组比较,SH-SY5Y细胞的存活率及凋亡率差异有统计学意义(P<0.05);相同浓度的苦参碱干预组与苦参碱联合顺铂干预组比较,SH-SY5Y细胞的存活率及凋亡率差异有统计学意义(P<0.05);苦参碱联合顺铂组与顺铂干预组比较,SH-SY5Y细胞的存活率及凋亡率差异有统计学意义(P<0.05)。苦参碱0.5 mg/ml组与顺铂干预组比较,SH-SY5Y细胞的存活率及凋亡率差异无统计学意义(P>0.05)。各组SH-SY5Y细胞间,p-TrkB的表达水平差异无统计学意义(P>0.05)。结论苦参碱可以抑制SH-SY5Y细胞的增殖并诱导凋亡,从而提高顺铂化疗的敏感性,该作用与TrkB/BDNF信号通路(间)无相关性。

Effect of matrine combined with cisplatin on drug sensitivity and TrkB expression in SH-SY5Y cells

Objective To explore the effects of matrine(Ma) on survival and apoptosis of neuroblastoma cell line SH-SY5Y,reduction of drug-resistance of cisplatin(CDDP) and its potential mechanism.Methods SH-SY5Y cells were treated with all-trans-retinoic acid(ATRA),brain-derived neurotrophic factor,CDDP and Ma respectively.Trials were divided into following groups: control group,ATRA induced cisplatin intervention group,cisplatin intervention group,Ma intervention group,and Ma combined with cisplatin intervention group.MTT colorimetric assay was performed to exam the survival rate of SH-SY5Y cells treated with Ma and CDDP.Apoptosis rate of SY5Y was measured by flow cytometry Ma.Western-blot analysis was employed to measure the p-TrkB expression.Results There was significant difference in survival and apoptosis of SH-SY5Y cells between each group and control group except the ATRA induced cisplatin intervention group(P < 0.05).Significant difference was also found between cisplatin intervention group and ATRA induced cisplatin intervention group(P < 0.05).The survival rates and apoptosis of SH-SY5Y cells was statistically significant between Ma group and Ma combined with cisplation group(P <0.05);and also between the Ma joint cisplation group and cisplation intervention group(P < 0.05);There was no significant difference in survival and apoptosis of SH-SY5Y cells between Ma(0.5 mg / ml) intervention group and ATRA induced cisplatin group(P > 0.05).No significant difference was observed in p-TrkB expression of SH-SY5Y cells among groups(P > 0.05).Conclusions Ma can induce cell apoptosis and inhibit the growth of the SH-SY5Y cells,meanwhile increase the sensibility of chemotherapy of cisplatin but this effect is not correlated to the TrkB /BDNF signal transduction.