Nakamura Toru, Torimura Takuji, Sakamoto Masaharu, Hashimoto Osamu, Taniguchi Eitaro, Inoue Kinya, Sakata Ryuichiro, Kumashiro Ryukichi, Murohara Toyoaki, Ueno Takato, Sata Michio
Division of Gastroenterology, Department of Medicine, Kurume University School of Medicine, Kurume, Fukuoka, Japan.
Gastroenterology. 2007 Jul;133(1):91-107.e1. doi: 10.1053/j.gastro.2007.03.110. Epub 2007 Apr 11.
BACKGROUND & AIMS: We investigated whether endothelial progenitor cell (EPC) transplantation could reduce established liver fibrosis and promote hepatic regeneration by isolating rat EPCs from bone marrow cells.
Recipient rats were injected intraperitoneally with carbon tetrachloride (CCl(4)) twice weekly for 6 weeks before initial administration of EPCs. CCl(4) was then readministered twice weekly for 4 more weeks, and EPC transplantation was carried out for these same 4 weeks.
At 7 days in culture, the cells expressed Thy-1, CD31, CD133, Flt-1, Flk-1, and Tie-2, suggesting an immature endothelial lineage. Immunohistochemical analyses showed fluorescent-labeled, transplantation EPCs were incorporated into the portal tracts and fibrous septa. Single and multiple EPC transplantation rats had reduced liver fibrosis, with decreased alpha2-(I)-procollagen, fibronectin, transforming growth factor-beta, and alpha-smooth muscle actin-positive cells. Film in situ zymographic analysis revealed strong gelatinolytic activity in the periportal area, in accordance with EPC location. Real-time polymerase chain reaction analysis of multiple EPC-transplantation livers showed significantly increased messenger RNA levels of matrix metalloproteinase (MMP)-2, -9 and -13, whereas tissue inhibitor of metalloproteinase-1 expression was significantly reduced. Expression of hepatocyte growth factor, transforming growth factor-alpha, epidermal growth factor, and vascular endothelial growth factor was increased in multiple EPC-transplantation livers, while hepatocyte proliferation increased. Transaminase, total bilirubin, total protein, and albumin levels were maintained in EPC-transplantation rats, significantly improving survival rates.
We conclude that single or repeated EPC transplantation halts established liver fibrosis in rats by suppressing activated hepatic stellate cells, increasing matrix metalloproteinase activity, and regulating hepatocyte proliferation.
我们通过从骨髓细胞中分离大鼠内皮祖细胞（EPC），研究其移植是否能减轻已形成的肝纤维化并促进肝再生。
在初次给予EPC前6周，每周两次给受体大鼠腹腔注射四氯化碳（CCl₄），共6周。之后继续每周两次给予CCl₄，持续4周，同时在这4周内进行EPC移植。
培养7天时，细胞表达Thy-1、CD31、CD133、Flt-1、Flk-1和Tie-2，提示为未成熟的内皮谱系。免疫组织化学分析显示，荧光标记的移植EPC整合到门管区和纤维间隔中。单次和多次EPC移植的大鼠肝纤维化减轻，α2-(I)-前胶原、纤连蛋白、转化生长因子-β和α-平滑肌肌动蛋白阳性细胞减少。膜原位酶谱分析显示，与EPC位置一致，门周区域有强烈的明胶酶活性。对多次EPC移植肝脏的实时聚合酶链反应分析显示，基质金属蛋白酶（MMP）-2、-9和-13的信使核糖核酸水平显著升高，而金属蛋白酶组织抑制剂-1的表达显著降低。多次EPC移植肝脏中肝细胞生长因子、转化生长因子-α、表皮生长因子和血管内皮生长因子的表达增加，同时肝细胞增殖增加。EPC移植大鼠的转氨酶、总胆红素、总蛋白和白蛋白水平保持稳定，存活率显著提高。
我们得出结论，单次或重复EPC移植可通过抑制活化的肝星状细胞、增加基质金属蛋白酶活性和调节肝细胞增殖来阻止大鼠已形成的肝纤维化。
