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探讨鱼藤酮对PC12细胞多巴胺转运体(DAT)和突触囊泡单胺转运体2(VMAT2)的影响。我们将PC12细胞经不同浓度的鱼藤酮处理24h后,利用HE染色观察PC12细胞的形态学变化;利用免疫组织化学法观察不同浓度的鱼藤酮对PC12细胞酪氨酸羟化酶(TH),DAT和VMAT2蛋白表达的影响;利用RT-PCR法检测鱼藤酮对PC12细胞DAT和VMAT2基因表达的影响;通过生化实验检测Na+/K+-ATP酶活性。实验结果显示:鱼藤酮浓度为1.0mmol/L时,PC12细胞出现了明显的形态学改变,胞浆染色不均、胞体呈圆形、细胞核形状不规则、核浆比例增加;TH蛋白阳性细胞数量减少;DAT蛋白和基因表达增加;VMAT2蛋白和基因表达降低,与对照组比较均具有统计学意义(P<0.05),且随着鱼藤酮浓度的增加,变化趋势更为明显。鱼藤酮浓度为1.0mmol/L时,Na+/K+-ATP酶活力与对照组比较显著降低(P<0.05)。因此,DAT和VMAT2的表达异常可能参与了毒性作用。
To investigate the effects of rotenone on dopamine transporter (DAT) and synaptic vesicle monoamine transporter 2 (VMAT2) in PC12 cells. PC12 cells were treated with different concentrations of rotenone for 24 h, then the morphological changes of PC12 cells were observed by HE staining. The changes of tyrosine hydroxylase (TH), DAT and VMAT2 in PC12 cells were observed by immunohistochemistry The effects of rotenone on the expression of DAT and VMAT2 genes in PC12 cells were detected by RT-PCR. The activity of Na + / K + -ATPase was detected by biochemical tests. The experimental results showed that when rotenone concentration was 1.0mmol / L, PC12 cells showed obvious morphological changes, cytoplasm staining was uneven, cell bodies were round, nuclear shape was irregular and the proportion of nuclear plasma increased; the number of TH protein positive cells decreased ; DAT protein and gene expression increased; VMAT2 protein and gene expression decreased, compared with the control group were statistically significant (P <0.05), and with the increase of rotenone concentration, the trend is more obvious. When the rotenone concentration was 1.0 mmol / L, the activity of Na + / K + -ATPase was significantly lower than that of the control group (P <0.05). Therefore, abnormal expression of DAT and VMAT2 may be involved in the toxic effect.