Rat hepatic stellate cell expression of α2-macroglobulin is a feature of cellular activation:: implications for matrix remodelling in hepatic fibrosis
Rat hepatic stellate cell expression of α2-macroglobulin is a feature of cellular activation:: implications for matrix remodelling in hepatic fibrosis
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DOI：
10.1042/cs19980020
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发表时间：
1998-08-01
期刊：
影响因子：
6
通讯作者：
Arthur, MJP
中科院分区：
文献类型：
--
作者：
Kawser, CA;Iredale, JP;Arthur, MJP
1.肝星状细胞是肝纤维化的关键介质。目的：1.研究肝星状细胞在组织培养中活化后胶原酶及其抑制物α2-巨球蛋白的表达，以及对某些细胞因子的反应。用Pronase-胶原酶消化法分离肝星状细胞，在未涂膜的塑料上培养活化。Northern分析显示，激活后(5天)的肝星状细胞表达α2-巨球蛋白mRNA，培养5~15天可见α2-巨球蛋白免疫定位于肝星状细胞。细胞培养上清液的酶联免疫吸附试验发现，肝星状细胞分泌α2-巨球蛋白由培养第5天的每24小时2.78+/-1.13 ng·ml(-1)增加到培养15天的每24 h 13.55+/-4.64 ng·ml(-1)。用白细胞介素6刺激肝星状细胞5天后，α2-巨球蛋白的表达显著增加，库普弗细胞条件培养液也是如此。然而，肝星状细胞对白介素1、转化生长因子-β1和肿瘤坏死因子-α的暴露没有显著影响。肝纤维化过程中，胆管结扎后(72 h~13 d)血浆α2-巨球蛋白水平升高至对照组(100%)的850%~250%之间，CCl(4)诱导的肝纤维化进展期(24 h~4周)血浆α2-巨球蛋白水平升高至对照组的1166%和1106%。这些数据表明，肝星状细胞是一种有效的蛋白酶抑制剂α2-巨球蛋白的潜在来源，其表达可能抑制进行性纤维化过程中的基质重塑。
1. Hepatic stellate cells are key mediators of hepatic fibrosis. We have studied hepatic stellate cell expression of the collagenase and general protease inhibitor alpha 2-macroglobulin after activation in tissue culture and in response to certain cytokines.2. Hepatic stellate cells isolated by Pronase-collagenase digestion were activated by culture on uncoated plastic. By Northern analysis hepatic stellate cells undergoing activation (5 days) expressed alpha 2-macroglobulin mRNA and alpha 2-macroglobulin could be immunolocalized to hepatic stellate cells from 5 to 15 days of culture.3. By ELISA of cell culture supernatants hepatic stellate cell secretion of alpha 2-macroglobulin was found to increase from 2.78 +/- 1.13 ng.ml(-1).mu g(-1) DNA per 24 h at 5 days of culture (n = 8) to 13.55 +/- 4.64 ng.ml(-1).mu g(-1) DNA per 24 h at 15 days of culture (n = 7). Stimulation of hepatic stellate cells with interleukin-6 at 5 days caused a significant increase in alpha 2-macroglobulin expression as did exposure to Kupffer-cell conditioned medium. However, exposure of hepatic stellate cells to interleukin-1, transforming growth factor-beta(1) and tumour necrosis factor-alpha had no significant effect.4. During profibrotic liver injury plasma alpha 2-macroglobulin levels were found to increase to between 850% and 250% of the control value (100%) after bile duct ligation (72 h to 13 days respectively), and to 1166% and 1106% of the control value during progressive CCl(4)-induced fibrosis (24 h to 4 weeks respectively).5. These data suggest that hepatic stellate cells are a potential source of the potent protease inhibitor alpha 2-macroglobulin, expression of which may inhibit matrix remodelling during progressive fibrosis.