右美沙芬对神经胶质瘤U87细胞的抑制作用及其机制研究  

Inhibitory Effect and Its Mechanism of Dextromethorphan on Glioma U87 Cell

作　　者：肖晶晶 毕逢辰 郑晓敏 王俊燕 李云鸿 王银 XIAO Jingjing;BI Fengchen;ZHENG Xiaomin;WANG Junyan;LI Yunhong;WANG Yin(Ningxia Key Laboratory of Cerebrocranial Diseases,Yinchuan 750004,China;Department of Neurobiology,School of Basic Medical Science,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏颅脑疾病重点实验室,银川750004 [2]宁夏医科大学基础医学院神经生物学系,银川750004

出　　处：《宁夏医科大学学报》2019年第10期973-976,共4页Journal of Ningxia Medical University

基　　金：国家自然科学基金（81571098;31560273）

摘　　要：目的探讨右美沙芬(dextromethorphan,DM)对胶质瘤U87细胞的抑制作用及其潜在机制。方法将神经胶质瘤U87细胞分别设为正常对照组及不同浓度DM实验组,利用CCK-8检测不同浓度DM对U87细胞活性的影响。Western blot检测DM处理48h后U87细胞中热休克蛋白60(HSP60)、热休克转录因子1(HSF-1)、半胱氨酸-天冬氨酸蛋白酶3(Caspase-3)、核因子-κB(NF-κB)和Toll样受体4(TLR-4)的表达水平。免疫荧光化学染色法检测DM处理48h后U87细胞的形态及数目变化。结果CCK-8细胞增殖实验检测结果表明,与对照组相比,10μM的DM处理U87细胞48 h后,细胞活力差异无统计学意义(P>0.05),而25~100μM的DM处理U87细胞后,细胞活力均降低(P<0.05或<0.01),与对照组比较,25μM的DM处理U87细胞48后HSP60、HSF-1和Caspase-3的表达升高,NF-κB和TLR-4表达均降低(P均<0.05)。结论DM对U87细胞活性具有抑制作用,但不是通过HSP60上调TLR-4/NF-κB信号通路进而促进U87细胞发生凋亡实现的,其机制有待进一步研究。Objective To investigate the effect of dextromethorphan(DM)on the proliferation of glioma U87 cells and its potential molecular mechanism.Methods U87 glioma cells were divided into normal control group and experimental group(U87+DM).CCK-8 cell proliferation assay was used to detect the influence of DM on the activity of U87 cells and to determine the optimal concentration of DM.The expression levels of heat shock protein60(HSP60),cysteine-aspartate protease 3(caspase-3),nuclear factor-κB(NF-κB)and toll-like receptor 4(TFR-4)in U87 cells were detected by Western blot.The expression levels of NF-κB and HSP60 in U87 cells treated with DM and the effects on morphology were detected by immunofluorescence staining.Results CCK-8 test results showed that compared with the control group,the activity of U87 cells treated with 25-100μM DM decreased(P<0.05),while U87 cells treated with 10μm DM for 48 h showed no effect on cell activity.With the increase of DM concentration,the proliferation of U87 cells was inhibited to varying degrees,and apoptosis was promoted.The expression of HSP60 and Caspase-3 were increased,while the expressions of NF-κB and TLR-4 were decreased.Conclusion DM has an inhibitory effect on U87 cell proliferation,but not through up regulating HSP60 initiated TLR-4/NF-κB signaling pathway to promote the apoptosis of U87 cells.The mechanism remains to be further studied.

关 键 词：右美沙芬 热休克蛋白60 核因子-ΚB TOLL样受体4 半胱氨酸-天冬氨酸蛋白酶3 神经胶质瘤 

分 类 号：R739.41[医药卫生—肿瘤]