目的：探索川芎嗪（ TMP）对WERI－Rb1细胞增殖的影响及TMP下调人WERI－Rb1细胞中趋化因子受体4（CXCR4）的转录的分子机制。方法（1）TMP及CXCR4的拮抗剂AMD3100药物分别处理WERI－Rb1细胞，相同浓度PBS及DMSO作为对照，活细胞成像仪观察WERI－Rb1细胞形态及密度变化。（2） WERI－Rb1细胞经TMP处理后，采用荧光定量逆转录PCR、免疫荧光及蛋白免疫印迹法分别从mRNA和蛋白质水平检测TMP对WERI －Rb1细胞 CXCR4及 NRF1表达的影响。结果（1）活细胞成像仪观察发现 TMP 处理组与AMD3100处理组的细胞状态较正常组差，增殖明显受抑制。（2）荧光定量逆转录PCR结果显示：与对照组比较， TMP处理组WERI－Rb1细胞中CXCR4和NRF1mRNA表达均明显下降（P＜0.05）；免疫荧光及蛋白免疫印迹实验结果显示，与对照组比较，TMP处理组WERI－Rb1细胞中CXCR4和NRF1蛋白表达均明显下降（ P＜0.05）。结论TMP对WERI－Rb1细胞的增殖有明显抑制作用，且能下调WERI－Rb1细胞中CXCR4及NRF1的表达水平。%Objective To characterize the bioactivity of tetramethylpyrazine ( TMP) on human WERI -Rb1 cells and its transcription regulatory mechanism of TMP -mediated down-regulation of CXCR4.Methods (1) WERI-Rb1 cells were treated with TMP and AMD3100.The control cells were treated with DMSO or PBS at the same concentration . The morphology and cell density of WERI -Rb1 cells was observed and recorded by the living cells imager .(2)WERI-Rb1 cells were treated with TMP;then real-time RT-PCR, immunofluorescence and western blot were performed to de-tect the expression levels of CXCR4 and NRF1 in WERI-Rb1 cells treated with TMP.Results (1)TMP and AMD3100 notably inhibited the proliferation of WERI -Rb1 cells, as evidenced by cell morphology and density analysis .(2)TMP down-regulated CXCR4 and NRF1 expression in WERI-Rb1 cells at both mRNA and protein levels (P<0.05), as ev-idenced by real-time RT-PCR, immunofluorescence and western blot assays .Conclusion TMP significantly inhibits the proliferation of WERI -Rb1 cells and down-regulates its expression of CXCR 4 and NRF1.
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