Department of Molecular and Cellular Medicine, Institute of Liver and Biliary Sciences (ILBS), New Delhi, 110 070, India.
Amity Institute of Molecular Medicine and Stem Cell Research, Amity University, Greater Noida, India.
Hepatol Int. 2021 Dec;15(6):1389-1401. doi: 10.1007/s12072-021-10244-0. Epub 2021 Aug 25.
Acute-on-chronic liver failure (ACLF) is a distinct clinical entity with high probability of organ failure and mortality. Since patients generally present late, experimental models are needed to understand the pathophysiology and natural course of the disease.
To reproduce the syndrome of ACLF, chronic liver disease was induced in C57BL6 mice (6-8 weeks; approximately 20-24 g weight) by intraperitoneal administration of carbon tetrachloride (CCl) for 10 weeks followed by an acute injury with acetaminophen (APAP) and lipopolysaccharide (LPS). Blood, ascitic fluid, and organs were collected to study cell death, regeneration, and fibrosis.
At 24 h post-APAP/LPS infusion, the liver tissue showed increased hepatocyte ballooning and endothelial cell TUNEL positivity. This was followed by progressive hepatocyte necrosis from perivascular region at day 7 to lobular region by day 11. ACLF (day 7 and day 11) animals showed increase in bilirubin (p < 0.05), prothrombin time (p < 0.0001), blood ammonia (p < 0.001), and portal pressure post-acute hepatocellular injury similar to human ACLF. Ascites was noticed by day 11 with median serum-ascites albumin gradient of 1.2 (1.1-1.3) g/dL. In comparison to cirrhosis, ACLF group (day 7 and day 11) showed significant decrease in Sirius red (p ≤ 0.0001), collagen1 (p < 0.0001), and a-SMA proportionate area (p < 0.0001) with loss of hepatocytes regeneration (p < 0.005). At day 11, ACLF animals also showed significant increase in serum creatinine (p < 0.05) and acute tubular necrosis suggestive of organ failure, compared to cirrhotic animals.
The CCL4/APAP/LPS (CALPS) model of ACLF mimics the clinical, biochemical, and histological features of ACLF with demonstrable progressive hepatocellular necrosis, liver failure, impaired regeneration, development of portal hypertension, and organ dysfunction in an animal with chronic liver disease.
慢加急性肝衰竭（ACLF）是一种具有高器官衰竭和死亡率概率的独特临床实体。由于患者通常就诊较晚，因此需要实验模型来了解疾病的病理生理学和自然病程。
为了再现 ACLF 的综合征，通过腹腔内给予四氯化碳（CCl）10 周来诱导 C57BL6 小鼠（6-8 周；约 20-24g 体重）的慢性肝病，然后用对乙酰氨基酚（APAP）和脂多糖（LPS）进行急性损伤。收集血液、腹水和器官以研究细胞死亡、再生和纤维化。
APAP/LPS 输注后 24 小时，肝组织显示出肝细胞气球样变和内皮细胞 TUNEL 阳性增加。随后，从第 7 天开始，从血管周围区域到第 11 天的小叶区域逐渐发生肝细胞坏死。ACLF（第 7 天和第 11 天）动物的胆红素（p<0.05）、凝血酶原时间（p<0.0001）、血氨（p<0.001）和急性肝细胞损伤后的门静脉压力均增加，类似于人类 ACLF。第 11 天出现腹水，中位血清腹水白蛋白梯度为 1.2（1.1-1.3）g/dL。与肝硬化相比，ACLF 组（第 7 天和第 11 天）的天狼星红（p≤0.0001）、胶原 1（p<0.0001）和α-SMA 比例面积（p<0.0001）显著减少，肝细胞再生减少（p<0.005）。第 11 天，与肝硬化动物相比，ACLF 动物的血清肌酐（p<0.05）也显著增加，提示发生器官衰竭的急性肾小管坏死。
CCL4/APAP/LPS（CALPS）ACLF 模型模拟了 ACLF 的临床、生化和组织学特征，具有可观察到的进行性肝细胞坏死、肝功能衰竭、再生受损、门静脉高压发展和慢性肝病动物的器官功能障碍。
