Fernández-Martínez Eduardo, Pérez-Alvarez Víctor, Tsutsumi Victor, Shibayama Mineko, Muriel Pablo
Centro de Investigación en Biología de la Reproducción, Universidad Autónoma del Estado de Hidalgo, Hidalgo, México.
Exp Toxicol Pathol. 2006 Aug;58(1):49-58. doi: 10.1016/j.etp.2006.03.002. Epub 2006 Apr 17.
Chronic bile duct ligation (BDL) is a useful model of cirrhosis. However, its parallel plasma and liver changes in levels of cytokines and nitric oxide (NO), involved in liver damage, remain unknown. The aims of this work were to quantify both the plasma and hepatic levels of five cytokines and NO in cirrhotic rats, 28 days after bile BDL, and to analyze their relationship with liver damage markers. One group of male Wistar rats was bile duct ligated and another group was sham operated, both groups were sacrificed 28 days after BDL. Plasma and liver cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin-6, -1beta, -10 (IL-6, -1beta, -10) and interferon-gamma (IFN-gamma), were measured by ELISA. Plasma and hepatic NO was determined as NO(2)(-)+NO(3)(-) by an enzymatic method. Alkaline phosphatase, gamma-glutamyl transpeptidase, alanine aminotransferase and bilirubins were determined in plasma. Collagen, lipid peroxidation and glycogen were quantified in liver. Two histopathological staining techniques were performed. BDL-induced cirrhosis was corroborated by the elevated liver damage markers and histopathological analysis. Chronic BDL significantly increased (P<0.05) most of plasma and hepatic cytokine levels and diminished the hepatic IFN-gamma amount. NO was increased in both tissues, but such change was only significant in plasma. Biliary cirrhosis produces interesting changes in plasma and hepatic levels of cytokines and NO. This finding in chronic BDL model in rats has not been previously described in both tissues for such cytokines and NO. Cytokines and NO imbalance favor establishment and perpetuation of cirrhosis.
慢性胆管结扎(BDL)是一种有用的肝硬化模型。然而,其在肝脏损伤中涉及的细胞因子和一氧化氮(NO)水平的血浆和肝脏平行变化仍不清楚。本研究的目的是量化胆管结扎28天后肝硬化大鼠血浆和肝脏中五种细胞因子及NO的水平,并分析它们与肝损伤标志物的关系。一组雄性Wistar大鼠进行胆管结扎,另一组进行假手术,两组在胆管结扎28天后处死。采用酶联免疫吸附测定法(ELISA)检测血浆和肝脏中的细胞因子,即肿瘤坏死因子-α(TNF-α)、白细胞介素-6、-1β、-10(IL-6、-1β、-10)和干扰素-γ(IFN-γ)。采用酶法将血浆和肝脏中的NO测定为NO₂⁻+NO₃⁻。测定血浆中的碱性磷酸酶、γ-谷氨酰转肽酶、丙氨酸氨基转移酶和胆红素。对肝脏中的胶原蛋白、脂质过氧化和糖原进行定量分析。进行了两种组织病理学染色技术。肝损伤标志物升高和组织病理学分析证实了BDL诱导的肝硬化。慢性BDL显著增加(P<0.05)了大多数血浆和肝脏细胞因子水平,并降低了肝脏中IFN-γ的含量。两种组织中的NO均增加,但这种变化仅在血浆中显著。胆汁性肝硬化在血浆和肝脏细胞因子及NO水平上产生了有趣的变化。大鼠慢性BDL模型中的这一发现此前在这两种组织中对于此类细胞因子和NO均未被描述过。细胞因子和NO失衡有利于肝硬化的建立和持续存在。