Regulation of CCN2 mRNA expression and promoter activity in activated hepatic stellate cells
Regulation of CCN2 mRNA expression and promoter activity in activated hepatic stellate cells
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DOI：
10.1007/s12079-008-0029-z
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发表时间：
2008-06-01
影响因子：
4.1
通讯作者：
Brigstock, David R.
中科院分区：
文献类型：
--
作者：
Leask, Andrew;Chen, Shaoqiong;Brigstock, David R.
基质细胞蛋白结缔组织生长因子（CCN 2）被认为是伤口愈合和纤维化中成纤维细胞活化的可靠标志物。CCN 2在肝星状细胞（HSC）活化期间被诱导。在这里，我们研究CCN 2基因在HSC中表达的分子基础。使用荧光激活细胞分选来研究CCl 4处理的小鼠体内HSC中CCN 2的表达。通过聚合酶链反应评估CCN 2和TGF-β mRNA表达作为培养诱导的HSC活化的函数。CCN 2启动子/报告基因构建体用于绘制基础和TGF β诱导的CCN 2启动子活性所需的顺式作用元件。实时聚合酶链反应分析用于进一步阐明HSC中CCN 2表达所需的信号通路。CCl 4体内给药增加HSC产生CCN 2。在体外，CCN 2和TGF-β mRNA的表达在培养的第0天至第14天之间在小鼠HSC中同时增加。TGF β诱导的CCN 2启动子活性需要CCN 2启动子中的Smad和Ets-1元件，并且通过TGF β I型受体（ALK 4/5/7）抑制而降低。活化的HSC中CCN 2过表达是ALK 4/5/7依赖性的。由于CCN 2过表达是纤维化发生的可靠标志物，我们的数据与通过TGF β I型受体如ALK 5的信号传导有助于HSC活化的观点一致，因此ALK 4/5/7抑制预期是肝纤维化的适当治疗。
The matricellular protein connective tissue growth factor (CCN2) is considered a faithful marker of fibroblast activation in wound healing and in fibrosis. CCN2 is induced during activation of hepatic stellate cells (HSC). Here, we investigate the molecular basis of CCN2 gene expression in HSC. Fluoroscence activated cell sorting was used to investigate CCN2 expression in HSC in vivo in mice treated with CCl4. CCN2 and TGF-beta mRNA expression were assessed by polymerase chain reaction as a function of culture-induced activation of HSC. CCN2 promoter/reporter constructs were used to map cis-acting elements required for basal and TGF beta-induced CCN2 promoter activity. Real-time polymerase chain reaction analysis was used to further clarify signaling pathways required for CCN2 expression in HSC. CCl4 administration in vivo increased CCN2 production by HSC. In vitro, expression of CCN2 and TGF-beta mRNA were concommitantly increased in mouse HSC between days 0 and 14 of culture. TGF beta-induced CCN2 promoter activity required the Smad and Ets-1 elements in the CCN2 promoter and was reduced by TGF beta type I receptor (ALK4/5/7) inhibition. CCN2 overexpression in activated HSC was ALK4/5/7-dependent. As CCN2 overexpression is a faithful marker of fibrogenesis, our data are consistent with the notion that signaling through TGF beta type I receptors such as ALK5 contributes to the activation of HSC and hence ALK4/5/7 inhibition would be expected to be an appropriate treatment for liver fibrosis.