Kupffer cells are associated with apoptosis, inflammation and fibrotic effects in hepatic fibrosis in rats
Kupffer cells are associated with apoptosis, inflammation and fibrotic effects in hepatic fibrosis in rats
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DOI：
10.1038/labinvest.2010.123
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发表时间：
2010-12-01
影响因子：
5
通讯作者：
Liu, Ping
中科院分区：
文献类型：
--
作者：
Liu, Cheng;Tao, Qing;Liu, Ping
肝细胞凋亡、肝脏炎症和纤维化是慢性肝病的显著特征。然而，这些过程之间的联系在机制上仍然不清楚。在这项研究中，我们研究了枯否细胞（KCs）的凋亡和激活，以及它们在肝纤维化过程中的病理生理作用。用二甲基亚硝胺（DMN）或四氯化碳（CCl_4）诱导大鼠肝纤维化。从正常大鼠中分离KCs并与脂多糖（LPS）或从纤维化大鼠中孵育。KC用抗-CD 68抗体（KC的生物标志物）化学染色。Western blot和实时荧光定量PCR检测CD 68的表达水平。应用共聚焦显微镜观察KCs的凋亡及其在肝纤维化形成中的病理生理作用。CD 68的mRNA和蛋白表达在DMN和CCL 4处理的大鼠中显著增加。共聚焦显微镜分析表明，CD 68阳性KC，但不是α-平滑肌肌动蛋白（SMA）阳性细胞，在DMN和CCL 4治疗大鼠的肝脏发生凋亡。此外，末端脱氧核苷酸转移酶介导的dUTP缺口末端标记和CD 68双阳性凋亡的KCs位于门静脉或纤维化间隔区，位于肝星状细胞（HSC）旁边。肿瘤坏死因子-α（TNF-α）和KC共定位于HSC附近的肝脏中。α-SMA和I型胶原双阳性细胞主要存在于纤维化隔中，这些细胞与CD 68阳性细胞明显共定位。有趣的是，在汇管区和肝窦中发现一些CD 68和Col（1）双阳性，但α-SMA完全阴性;这种现象也在LPS暴露6 h后的原代分离KC或体外纤维化大鼠中得到验证。这些结果表明，在肝纤维化模型中，KCs与肝细胞凋亡、炎症和纤维化过程相关。实验室调查（2010）90，1805-1816; doi：10.1038/labinvest.2010.123; 2010年10月4日在线发表
Hepatocellular apoptosis, hepatic inflammation, and fibrosis are prominent features in chronic liver diseases. However, the linkage among these processes remains mechanistically unclear. In this study, we examined the apoptosis and activation of Kupffer cells (KCs) as well as their pathophysiological involvement in liver fibrosis process. Hepatic fibrosis was induced in rats by dimethylnitrosamine (DMN) or carbon tetrachloride (CCl4) treatment. KCs were isolated from normal rats and incubated with lipopolysaccharide (LPS) or from fibrotic rats. The KCs were stained immunohistochemically with anti-CD68 antibody, a biomarker for KC. The level of expression of CD68 was analyzed by western blot and real-time PCR methods. The apoptosis and pathophysiological involvement of KCs in the formation of liver fibrosis were studied using confocal microscopy. The mRNA and protein expression of CD68 were significantly increased in DMN- and CCL4-treated rats. Confocal microscopy analysis showed that CD68-positive KCs, but not alpha-smooth muscle actin (SMA)-positive cells, underwent apoptosis in the liver of DMN- and CCL4-treated rats. It was also revealed that the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling and CD68-double-positive apoptotic KCs located in the portal or fibrotic septa area were situated next to hepatic stellate cells (HSCs). Tumor necrosis factor-alpha (TNF-alpha) and KC co-localized in the liver in the neighbor of HSCs. The double alpha-SMA- and collagen type I-positive cells predominantly existed in fibrotic septa, and those cells were co-localized clearly with CD68-positive cells. Interestingly, some CD68 and Col (1) double positive, but completely negative for alpha-SMA, were found in the portal areas and hepatic sinusoids; this phenomenon was also validated in primary isolated KCs after 6 h LPS exposure or fibrotic rats in vitro. These results show that KCs are associated with hepatocellular apoptosis, inflammation, and fibrosis process in a liver fibrosis models. Laboratory Investigation (2010) 90, 1805-1816; doi:10.1038/labinvest.2010.123; published online 4 October 2010