目的:使用四氯化碳(CCl4)和D-半乳糖胺(D-GalN)的实验模型可用于急性肝衰竭(ALF)治疗的临床前评估。不幸的是,这些模型的特点是不同的动态的肝损伤取决于动物品系,服用肝毒素,和它的剂量。这项研究的目的是比较已知的ALF大鼠和小鼠模型,以期将其引入临床前细胞治疗实验。特别是,根据组织病理学和分子变化,我们提出了有效干细胞治疗干预的实验时间截止点.
方法:通过在小鼠(50μL/100gb.w.)和大鼠(200μL/100gb.w.)以及在小鼠(150mg/100gb.w.)和大鼠(50mg/100gb.w.)中单次腹膜内注射CCl4诱导ALF。收集血液和肝脏样本12小时,24h,中毒后48小时和7天。血液形态学,肝功能血液检查,组织病理学变化,增殖活性,凋亡,纤维化,分析和基因表达以评估肝损伤。
结果:在12小时,24h,CCl4注入后48小时,小鼠肝脏显示中度炎症浸润和大量中央周围坏死。在用CCl4治疗的大鼠中,在12小时时观察到肝实质中的少量淋巴细胞浸润,随后在24小时出现中央静脉周围的坏死,并持续到48小时。在注射D-GalN的小鼠中,肝损伤的第一个组织病理学征象出现在48小时。在D-GalN处理的大鼠的肝脏中,12小时后发生中度中央周围炎性浸润,24h,48小时,伴随着增殖和凋亡的增加。所有组织学变化都伴随着某些基因表达的降低。在大多数实验组的大鼠和小鼠中,在中毒后48小时至7天之间,组织学和分子参数均恢复到基线值。
结论:在CCl4诱导的ALF的小鼠和大鼠中,肝功能衰竭的迹象可以在12小时内观察到,发展到48小时。在D-GalN诱导的模型中,小鼠对肝毒性作用的抵抗力高于大鼠(12小时后),肝炎的早期阶段可以观察到很晚,48小时后。这些截止点似乎是抑制炎症和应用有效的干细胞治疗急性肝损伤的最佳选择。
OBJECTIVE: Experimental models using carbon tetrachloride (CCl4) and D-galactosamine (D-GalN) can be used in preclinical assessment of acute liver failure (ALF) therapies. Unfortunately, these models are characterized by different dynamics of liver injury depending on the animal strain, administered hepatotoxin, and its dose. The aim of this study was to compare known rat and mouse models of ALF with a view to their future introduction into preclinical cell therapy experiments. In particular, based on histopathological and molecular changes, we suggested experimental time cut-off points for an effective stem cell therapeutic intervention.
METHODS: ALF was induced by a single intraperitoneal injection of CCl4 in mice (50 μL/100 g b.w.) and rats (200 μL/100 g b.w.) and D-GalN in mice (150 mg/100 g b.w.) and rats (50 mg/100 g b.w.). Blood and liver samples were collected 12 h, 24 h, 48 h and 7 days after intoxication. Blood morphology, liver function blood tests, histopathological changes, proliferation activity, apoptosis, fibrosis, and gene expression were analysed to assess liver damage.
RESULTS: At 12 h, 24 h, and 48 h after CCl4 injection, mouse livers showed moderate inflammatory infiltration and massive pericentral necrosis. In rats treated with CCl4, minor lymphocytic infiltration in the liver parenchyma was seen at 12 h, followed by necrosis that appeared around central veins at 24 h and persisted to 48 h. In D-GalN-injected mice, the first histopathological signs of liver injury appeared at 48 h. In the livers of D-GalN-treated rats, moderate pericentral inflammatory infiltration occurred after 12 h, 24 h, and 48 h, accompanied by increased proliferation and apoptosis. All histological changes were accompanied by decreasing expression of certain genes. In most experimental groups of rats and mice, both histological and molecular parameters returned to the baseline values between 48 h and 7 days after intoxication.
CONCLUSIONS: In mice and rats with CCl4-induced ALF, signs of liver failure can be seen as early as 12 h and develop to 48 h. In the D-GalN-induced model, mice are more resistant to the hepatotoxic effect than rats (after 12 h), and the early hepatitis phase can be observed much later, after 48 h. These cut-off points seem to be optimal for suppressing inflammation and applying effective stem cell therapy for acute liver injury.