Institute of Biomedicine (IBIOMED), University of León, León, Spain; Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd), Spain.
Institute of Biomedicine (IBIOMED), University of León, León, Spain.
Transl Res. 2015 Feb;165(2):346-57. doi: 10.1016/j.trsl.2014.10.003. Epub 2014 Oct 13.
We investigated whether melatonin ameliorates fibrosis and limits the expression of fibrogenic genes in mice treated with carbon tetrachloride (CCl4). Mice in treatment groups received CCl4 5 μL/g body weight intraperitoneally twice a week for 4 or 6 weeks. Melatonin was given at 5 or 10 mg/kg/d intraperitoneally, beginning 2 weeks after the start of CCl4 administration. Treatment with CCl4 resulted in fibrosis evidenced by the staining of Van Gieson and α-smooth muscle actin (α-SMA) positive cells in the liver. At both 4 and 6 weeks, CCl4 induced an increase in the messenger RNA levels of collagens I and III, transforming growth factor (TGF)-β, platelet-derived growth factor (PDGF), connective tissue growth factor (CTGF), amphiregulin, matrix metalloproteinase (MMP)-9, and tissue inhibitor of metalloproteinase (TIMP)-1. Protein concentrations of CTGF, amphiregulin, MMP-9, TIMP-1, and phospho-Smad3 were also significantly augmented in fibrotic mice. Melatonin successfully attenuated liver injury, as shown by histopathology and decreased levels of serum transaminases. Immunohistochemical staining of α-SMA indicated an abrogation of hepatic stellate cell activation by the indol. Furthermore, melatonin treatment resulted in significant inhibition of the expression of collagens I and III, TGF-β, PDGF, CTGF, amphiregulin, and phospho-Smad3. The MMP-9 activity decreased and the expression of nuclear factor erythroid-2-related factor 2 (Nrf2) increased in mice receiving melatonin. Data obtained suggest that attenuation of multiple profibrogenic gene pathways contributes to the beneficial effects of melatonin in mice with CCl4-induced liver fibrosis.
我们研究了褪黑素是否可以改善四氯化碳(CCl4)处理小鼠的纤维化,并限制成纤维基因的表达。治疗组小鼠每周两次腹膜内注射 5μL/g 体重的 CCl4,共 4 或 6 周。从 CCl4 给药开始 2 周后,开始腹膜内给予褪黑素 5 或 10mg/kg/d。CCl4 处理导致肝脏中 Van Gieson 和 α-平滑肌肌动蛋白(α-SMA)阳性细胞染色证实的纤维化。在 4 和 6 周时,CCl4 诱导 I 型和 III 型胶原、转化生长因子(TGF)-β、血小板衍生生长因子(PDGF)、结缔组织生长因子(CTGF)、 Amphiregulin、基质金属蛋白酶(MMP)-9 和金属蛋白酶组织抑制剂(TIMP)-1 的信使 RNA 水平增加。在纤维化小鼠中,CTGF、Amphiregulin、MMP-9、TIMP-1 和磷酸化 Smad3 的蛋白浓度也显著升高。褪黑素通过组织病理学和降低血清转氨酶水平成功减轻肝损伤。α-SMA 的免疫组织化学染色表明,该吲哚可以阻止肝星状细胞的活化。此外,褪黑素治疗导致 I 型和 III 型胶原、TGF-β、PDGF、CTGF、Amphiregulin 和磷酸化 Smad3 的表达显著抑制。MMP-9 活性降低,接受褪黑素治疗的小鼠核因子红细胞 2 相关因子 2(Nrf2)表达增加。研究结果表明,抑制多种致纤维化基因途径有助于褪黑素在 CCl4 诱导的肝纤维化小鼠中发挥有益作用。