College of Life Science, Henan Normal University, Xinxiang 453007, People’s Republic of China.
Cell Biol Int. 2012 Aug 1;36(8):721-32. doi: 10.1042/CBI20110104.
KCs (Kupffer cells), as an important hepatic immunoregulatory cells, play a key role in LR (liver regeneration). Uncovering the transcriptional profiling of KCs after PH (partial hepatectomy) would likely clarify its implication in LR. Here, we isolated KCs by methods of Percoll density gradient centrifugation and immunomagnetic beads. Transcriptional profiles of KCs were monitored up to 168 h post-PH using microarray. By comparing the expression profile of KCs at 2-168 h post-PH with that of the control and applying the statistical and bioinformatics criteria, we found 1407 known and 927 unknown genes related to LR. K-means clustering analysis grouped these 1407 genes into robust 14 time-course clusters representing distinct patterns of regulation. Based on gene-set enrichment analysis, genes encoding products involved in cytokine signalling, inflammatory response and cell chaemotaxis were highly enriched in the cluster characterized by gradual up-regulation and then return; genes in defence response and immune response were enriched in clusters 'the general down-regulation during LR'; genes in fatty acid synthesis and sterol metabolism were preferentially distributed in the cluster 'gradual increase'; whereas genes in the categories 'lipid catabolism' and 'glycolysis' were enriched in cluster 'decrease at two intervals'. According to the above analysis, KCs were seemingly sensitive to operative stimulus; immune defence and detoxification function of KCs obviously dropped post-operatively; fatty acid synthesis were enhanced, whereas lipid catabolism and glycolysis were reduced after PH. This study provides a detailed in vivo gene expression profile of KCs, providing a framework to better understand the molecular mechanisms underlying the regeneration process at cellular level.
KCs（库普弗细胞）作为一种重要的肝免疫调节细胞，在 LR（肝再生）中起着关键作用。揭示 PH（部分肝切除术）后 KCs 的转录谱特征可能阐明其在 LR 中的作用。在这里，我们通过 Percoll 密度梯度离心和免疫磁珠分离方法分离 KCs。使用微阵列监测 PH 后 2-168 小时 KCs 的转录谱。通过比较 PH 后 2-168 小时 KCs 的表达谱与对照，并应用统计和生物信息学标准，我们发现了 1407 个已知和 927 个与 LR 相关的未知基因。K-均值聚类分析将这些 1407 个基因分为 14 个稳健的时间过程簇，代表了不同的调节模式。基于基因集富集分析，编码细胞因子信号、炎症反应和细胞趋化性产物的基因在逐渐上调然后恢复的特征簇中高度富集；防御反应和免疫反应相关基因在 LR 中普遍下调的簇中富集；脂肪酸合成和固醇代谢相关基因优先分布在逐渐增加的簇中；而脂质分解和糖酵解相关基因在两个时间段下降的簇中富集。根据上述分析，KCs 似乎对手术刺激敏感；术后 KCs 的免疫防御和解毒功能明显下降；PH 后脂肪酸合成增强，而脂质分解和糖酵解减少。本研究提供了 KCs 的详细体内基因表达谱，为更好地理解细胞水平再生过程的分子机制提供了框架。
