Comparative analysis of expression profiles of chemokines, chemokine receptors, and components of signaling pathways mediated by chemokines in eight cell types during rat liver regeneration
Comparative analysis of expression profiles of chemokines, chemokine receptors, and components of signaling pathways mediated by chemokines in eight cell types during rat liver regeneration
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作者：
Chen, Xiaoguang;Xu, Cunshuan;Ma, Ji
已有研究表明，趋化因子可以在组织水平上积极调节部分肝切除术后的肝再生。然而，趋化因子在细胞水平上对再生的作用的确切机制仍然不清楚。本研究采用Percoll密度梯度离心法和免疫磁珠法分离纯化了2/3肝切除后8个恢复时间点的大鼠再生肝中的8种细胞。使用微阵列监测每种细胞类型的表达谱。RT-PCR分析验证芯片结果的可靠性。结果表明，总体上，趋化因子和受体基因的表达谱在不同的细胞类型之间存在差异;大多数参与趋化因子信号通路的基因在肝再生过程中在8种肝细胞类型中表达增加。利用生物信息学和系统生物学方法分析了这些基因在再生中的作用。根据基因芯片结果和基因协同作用，CCL 2-CCR 2和CCL 7-CCR 3可能分别激活胆管上皮细胞24 h和树突状细胞2-12 h的趋化因子信号通路，CCL 7-CCR 1可能激活肝窦内皮细胞2-12 h的Plc/Pkc和Pi 3 k/Akt信号通路;启动阶段Src/Ptk、Src/Vav和Plc/Pkc通路的激活可能与CCL 7的诱导作用有关。这些数据表明促炎趋化因子在细胞水平上与肝再生的潜在相关性。
It has been documented that chemokines can positively regulate liver regeneration at the tissue level after partial hepatectomy. However, the precise mechanism of the effects of chemokines on regeneration at the cellular level remains poorly defined. In this study, 8 cell types from rat regenerating liver at 8 recovery time points after 2/3 hepatectomy were isolated and purified using Percoll density gradient centrifugation and immunomagnetic bead methods. The expression profiles of each cell type were monitored using a microarray. RT-PCR analysis was performed to validate the reliability of the microarray results. The results showed that, on the whole, the expression profiles of chemokine and receptor genes varied among different cell types; most genes involved in chemokine signaling pathways showed an increase in expression across the 8 liver cell types during liver regeneration. The implication of these genes in regeneration was analyzed by bioinformatics and systems biology methods. According to the microarray results and gene synergy, activation of chemokine signaling pathways at 24 h in biliary epithelial cells and at 2-12 h in dendritic cells may be triggered by CCL2-CCR2 and CCL7-CCR3, respectively; activation of Plc/Pkc and Pi3k/Akt pathways at 2-12 h in sinusoidal endothelial cells might be caused by CCL7-CCR1; and activation of the Src/Ptk, Src/Vav, and Plc/Pkc pathways at the priming stage may be related to the inductive effect of CCL7. These data suggest the potential relevance of the pro-inflammatory chemokines for liver regeneration at the cellular level.