和厚朴酚对组织因子途径抑制物 2诱导神经胶质瘤细胞凋亡的实验研究#br#

目的探讨和厚朴酚对组织因子途径抑制物 2（TFPI 2）表达的影响，以及和厚朴酚和TFPI 2过表达对人胶质瘤U251细胞凋亡的影响。方法体外培养胶质瘤U87、U251、LN18、LN229、A172细胞系，荧光定量PCR（QPCR）检测不同细胞系中TFPI 2的表达。不同浓度（0、10、20、30、40和50 μmol／L）和厚朴酚干预U251细胞，QPCR和Western blotting检测TFPI 2 mRNA和蛋白的表达。采用腺病毒载体pGSadeno TFPI 2过表达U251细胞内TFPI 2的表达，流式细胞术及caspase 3试剂盒检测细胞凋亡率和caspase 3活性。结果在选取的5种不同胶质瘤细胞株中，U251和A172细胞内TFPI 2 mRNA明显下降（P＜005）。和厚朴酚呈浓度依赖性增加U251细胞内TFPI 2 mRNA水平；其中50 μmol／L和厚朴酚干预24 h后， TFPI 2 mRNA表达水平升高最明显（P＜005）。腺病毒感染U251细胞后TFPI 2表达水平明显增加（P＜005）。过表达TFPI 2和50 μmol／L和厚朴酚干预均可增加U251细胞凋亡水平和caspase 3的活性（P＜005）。结论和厚朴酚处理可增加U251细胞内TFPI 2的表达；和厚朴酚联合TFPI 2过表达显著诱导胶质瘤细胞凋亡。

Experimental study of honokiol on apoptosis of glioma cells induced by tissue factor pathway inhibitor 2#br##br#

ObjectiveTo investigate the effect of honokiol on the expression of tissue factor pathway inhibitor 2（TFPI 2） and the effect of overexpression of honokiol and TFPI 2 on the apoptosis of human glioma U251 cells. MethodsThe U87， U251， LN18， LN229 and A172 cell lines of glioma were cultured in vitro， and the expression of TFPI 2 in different cell lines was detected by fluorescence quantitative PCR （QPCR）. Honokiol treated U251 cells with different concentrations （0， 10， 20， 30， 40 and 50 μmol／L）. QPCR and Western blotting were used to detect the expression changes of TFPI 2 mRNA and protein. The expression of TFPI 2 in U251 cells was overexpressed by adenovirus vector pGSadeno TFPI 2， and apoptosis rate and caspase 3 activity were detected by flow cytometry and caspase 3 kit. ResultsThe TFPI 2 mRNA in U251 and A172 cells decreased significantly in U87， U251， LN18， LN229 and A172 cell lines （P＜005）. Honokiol increased TFPI 2 mRNA level in U251 cells in a concentration dependent manner （P＜005）. After 50 μmol／L honokiol intervened for 24 h， the expression level of TFPI 2 mRNA increased most significantly （P＜005）. The expression level of TFPI 2 in U251 cells increased significantly after transfected with adenovirus vector pGSadeno TFPI 2 （P＜005）. Overexpression of TFPI 2 and 50 μmol／L honokiol could increase U251 cell apoptosis and caspase 3 activity （P＜005）. ConclusionHonokiol can increase the expression of TFPI 2 in U251 cells， and overexpressed TFPI 2 combined with honokiol significantly induced apoptosis in glioma cells.