大鼠肝纤维化形成过程中肝组织细胞外信号调节激酶的动态表达

Dynamic expression of extracellular signal-regulated kinase in rat liver tissue during hepatic fibrogenesis.

作者信息

Zhang Xiao-Lan, Liu Jin-Ming, Yang Chang-Chun, Zheng Yi-Lin, Liu Li, Wang Zhan-Kui, Jiang Hui-Qing

机构信息

Department of Gastroenterology, the Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei Province, China.

出版信息

World J Gastroenterol. 2006 Oct 21;12(39):6376-81. doi: 10.3748/wjg.v12.i39.6376.

DOI:10.3748/wjg.v12.i39.6376

PMID:17072965

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4088150/

Abstract

AIM

To investigate whether extracellular signal-regulated kinase 1 (ERK(1)) is activated and associated with hepatic stellate cell (HSC) proliferation in fibrotic rat liver tissue.

METHODS

Rat hepatic fibrosis was induced by bile duct ligation (BDL). Histopathological changes were evaluated by hematoxylin and eosin staining, and Masson's trichrome method. ERK(1) mRNA in rat liver tissue was determined by reverse transcription-polymerase chain reaction, while the distribution of ERK(1) was assessed by immunohistochemistry. ERK(1) protein was detected by Western blotting analysis. The number of activated HSCs was quantified after alpha smooth muscle actin (alpha-SMA) staining.

RESULTS

With the development of hepatic fibrosis, the positive staining cells of alpha-SMA increased obviously, and mainly resided in the portal ducts. Fiber septa and perisinuses were accompanied with proliferating bile ducts. The positive staining areas of the rat livers in model groups 1-4 wk after ligation of common bile duct (12.88% +/- 2.63%, 22.65% +/- 2.16%, 27.45% +/- 1.86%, 35.25% +/- 2.34%, respectively) were significantly larger than those in the control group (5.88% +/- 1.46%, P < 0.01). With the development of hepatic fibrosis, the positive cells of ERK(1) increased a lot, and were mainly distributed in portal ducts, fiber septa around the bile ducts, vascular endothelial cells and perisinusoidal cells. Western blotting analysis displayed that the expression of ERK(1) and ERK(2) protein was up-regulated during the model course, and its level was the highest 4 wk after operation, being 3.9-fold and 7.2-fold higher in fibrotic rat liver than in controls. ERK(1) mRNA was expressed in normal rat livers as well, which was up-regulated two days after BDL and reached the highest 4 wk after BDL. The expression of ERK(1) was positively correlated with alpha-SMA expression (r = 0.958, P < 0.05).

CONCLUSION

The expression of ERK(1) protein and mRNA is greatly increased in fibrotic rat liver tissues, which may play a key role in HSC proliferation and hepatic fibrogenesis.

摘要

目的

研究细胞外信号调节激酶1（ERK(1)）在纤维化大鼠肝组织中是否被激活以及与肝星状细胞（HSC）增殖的关系。

方法

采用胆管结扎（BDL）诱导大鼠肝纤维化。通过苏木精-伊红染色和Masson三色染色法评估组织病理学变化。采用逆转录-聚合酶链反应测定大鼠肝组织中ERK(1) mRNA，免疫组织化学法评估ERK(1)的分布。通过蛋白质印迹分析检测ERK(1)蛋白。α平滑肌肌动蛋白（α-SMA）染色后对活化的HSCs数量进行定量。

结果

随着肝纤维化的发展，α-SMA阳性染色细胞明显增多，主要位于门静脉。纤维间隔和肝血窦周围伴有增生的胆管。胆总管结扎后1 - 4周模型组大鼠肝脏的阳性染色面积（分别为12.88%±2.63%、22.65%±2.16%、27.45%±1.86%、35.25%±2.34%）显著大于对照组（5.88%±1.46%，P < 0.01）。随着肝纤维化的发展，ERK(1)阳性细胞大量增加，主要分布在门静脉、胆管周围纤维间隔、血管内皮细胞和肝血窦周围细胞。蛋白质印迹分析显示，在模型过程中ERK(1)和ERK(2)蛋白表达上调，术后4周其水平最高，纤维化大鼠肝脏中的水平比对照组高3.9倍和7.2倍。ERK(1) mRNA在正常大鼠肝脏中也有表达，BDL后2天上调，BDL后4周达到最高。ERK(1)的表达与α-SMA表达呈正相关（r = 0.958，P < 0.05）。