Gene expression profiles during hepatic stellate cell activation in culture and in vivo
Gene expression profiles during hepatic stellate cell activation in culture and in vivo
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DOI：
10.1053/j.gastro.2007.02.033
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发表时间：
2007-05-01
期刊：
影响因子：
29.4
通讯作者：
Schwabe, Robert F.
中科院分区：
文献类型：
--
作者：
De Minicis, Samuele;Seki, Ekihiro;Schwabe, Robert F.
背景与目的：肝损伤后，肝星状细胞（HSCs）可转分化为生成细胞外基质的肌成纤维细胞，促进肝纤维化。在本研究中，我们测定了3种不同的HSC活化模型中基因表达的变化，并研究了HSC培养活化是否再现了HSC体内活化的基因表达变化。方法：采用密度离心和磁性抗体细胞分选分离正常小鼠、ccl4处理小鼠和胆管结扎小鼠的造血干细胞。基因表达通过芯片分析，聚合酶链反应和Western blot分析证实。结果：2373个探针组在3种HSC激活模型中至少1种中存在差异表达，包括编码促炎和抗凋亡介质的新基因；转录因子;细胞表面受体；以及细胞骨架成分，如CXCL14、survivin、septin 4、骨桥蛋白、PRX1、LMCD1、GPR91、leomodin和氨酰胺。BDL-和ccl4激活的造血干细胞显示出高度相关的基因表达模式，而培养激活仅部分再现了BDL-和ccl4诱导激活时观察到的基因表达变化。在培养激活过程中，与Kupffer细胞共培养或脂多糖处理将大多数检测基因的表达转向体内激活过程中观察到的模式，这表明这些因素在驱动HSC激活的微环境中发挥作用。结论：BDL和CCl4治疗后的HSC基因表达模式几乎相同，表明HSC在不同类型的肝损伤中发挥相似的功能。由于培养激活不能正确调节HSC中的基因表达，因此体内激活应被视为研究HSC生物学的金标准。
Background & Aim: Following hepatic injury, hepatic stellate cells (HSCs) transdifferentiate to become extracellular matrix-producing myofibroblasts and to promote hepatic fibrogenesis. In this study, we determine gene expression changes in 3 different models of HSC activation and investigate whether HSC culture activation reproduces gene expression changes of HSC in vivo activation. Methods: HSCs were isolated by density centrifugation and magnetic antibody cell sorting from normal mice, CCl4-treated mice, and mice that underwent bile duct ligation (BDL). Gene expression was analyzed by microarray and confirmed by polymerase chain reaction and Western blot analysis. Results: Two thousand seventy-three probe sets were differentially expressed in at least 1 of 3 models of HSC activation, including novel genes that encode proinflammatory and antiapoptotic mediators; transcription factors; cell surface receptors; and cytoskeleton components such as CXCL14, survivin, septin 4, osteopontin, PRX1, LMCD1, GPR91, leiomodin, and anillin. BDL- and CCl4-activated HSCs showed highly correlated gene expression patterns, whereas culture activation only partially reproduced the gene expression changes observed during BDL- and CCl4-induced activation. Coculture with Kupffer cells or lipopolysaccharide treatment during culture activation shifted the expression of most examined genes toward the pattern observed during in vivo activation, suggesting a role for these factors in the microenvironment that drives HSC activation. Conclusions: The almost identical HSC gene expression patterns after BDL or CCl4 treatment indicate that HSCs exert similar functions in different types of liver injury. Because culture activation does not properly regulate gene expression in HSCs, in vivo activation should be considered the gold standard for the study of HSC biology.