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培养的大鼠肝细胞中I型和IV型前胶原基因表达

Types I and IV procollagen gene expression in cultured rat hepatocytes.

作者信息

Clement B, Laurent M, Guguen-Guillouzo C, Lebeau G, Guillouzo A

机构信息

INSERM U-49, Unité de Recherches Hépatologiques, Hôpital de Pontchaillou, Rennes, France.

出版信息

Coll Relat Res. 1988 Jul;8(4):349-59. doi: 10.1016/s0174-173x(88)80006-2.

Abstract

The molecular mechanism involved in the expression of collagens by hepatocytes were investigated in both pure and co-culture with another rat liver epithelial cell type (RLEC). We measured the steady-state levels of mRNAs coding for pro alpha 1(I) and pro alpha 1(IV) chains by Northern analysis and by dot blotting, using specific recombinant cDNA probes. In freshly isolated hepatocytes, only small amounts of pro alpha 1(I) and pro alpha 1(IV) mRNAs were detected by dot-blot analysis. After 3 days in culture, the pro alpha 1(I) and pro alpha 1(IV) mRNA levels increased 2 to 5 times. The amount of pro alpha 1(IV) mRNAs was identical in hepatocyte cultured with RLECs while the pro alpha 1(I) mRNA level was 5 times that in pure hepatocyte culture. Hydrocortisone reduced pro alpha 1(I) mRNA in hepatocyte cultures, but had no effect on co-cultured cells. In both culture systems, this glucocorticoid did not act on the steady-state pro alpha 1(IV) mRNA level. Whatever the age and the type of culture (pure or mixed) RLECs exhibited the highest levels of pro alpha 1(I) and pro alpha 1(IV) mRNAs, which were reduced by hydrocortisone. These results show that procollagen gene expression by hepatocytes is not directly correlated with their functional state and that corticosteroids differently affect the expression of different collagen genes and collagen deposition.

摘要

我们研究了肝细胞表达胶原蛋白所涉及的分子机制,实验采用了肝细胞单独培养以及与另一种大鼠肝上皮细胞类型(RLEC)共培养的方式。我们使用特异性重组cDNA探针,通过Northern印迹分析和斑点印迹法,测量了编码前α1(I)和前α1(IV)链的mRNA的稳态水平。在新鲜分离的肝细胞中,通过斑点印迹分析仅检测到少量的前α1(I)和前α1(IV)mRNA。培养3天后,前α1(I)和前α1(IV)mRNA水平增加了2至5倍。与RLEC共培养的肝细胞中前α1(IV)mRNA的量与单独培养的肝细胞相同,而前α1(I)mRNA水平是单独培养肝细胞中的5倍。氢化可的松降低了肝细胞培养物中前α1(I)mRNA的水平,但对共培养细胞没有影响。在两种培养系统中,这种糖皮质激素对前α1(IV)mRNA的稳态水平均无作用。无论RLEC的年龄和培养类型(单独培养或混合培养)如何,其前α1(I)和前α1(IV)mRNA水平均最高,且氢化可的松可使其降低。这些结果表明,肝细胞中前胶原基因的表达与其功能状态没有直接相关性,并且皮质类固醇对不同胶原基因的表达和胶原沉积有不同的影响。

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