• Modeling Mechanism：

  N-Nitrosodiethylamine (DEN), a classic N-nitroso compounds that are liver carcinogens, is activated by hepatic cytochrome P450 enzymes after entering the rat body, producing an alkylating intermediate. This intermediate can bind to hepatocyte DNA to form adducts, leading to base damage and gene mutations. It also triggers oxidative stress (accumulation of reactive oxygen species, ROS), activates the nuclear factor-κB (NF-κB) signaling pathway, promotes the release of pro-inflammatory factors (TNF-α, IL-6), and induces hepatocyte inflammation, necrosis, and abnormal proliferation, gradually progressing to liver fibrosis, hepatocellular adenoma, and ultimately hepatocellular carcinoma (HCC), mimicking the complete pathological process of chemically induced liver tumors in humans.

• Related Products：

  N-Nitrosodiethylamine (T35455)

• Modeling Method：

  Experimental Subject：

  Rats, Male, Body weight 200–250 g, Age 3–4 months

  Dosage and Administration Route：

  ① Core modelling: N-Nitrosodiethylamine, 200 mg/kg, dissolved in sterile saline, administered intraperitoneally;
  ② Promoter combination (optional): Post-modelling, combine with phenobarbital (50 mg/kg/day, mixed into feed) to accelerate liver tumour progression;
  ③ Control treatment: Control group administered 10% Tween 80 solution (oral) or equal volume saline (intraperitoneal injection), with all other housing and treatment conditions identical;
  ④ Intervention validation group (optional): Administer hepatoprotective agent (e.g., 5-benzylidene-2-thioethanamide) concurrently with modelling , 50 mg/kg, Oral administration, Continuous for 3–6 weeks

  Dosing Frequency and Duration Model：

  Single dose

• Validation：

  1. Biochemical Indicators: - Liver Function: 3-6 weeks after modeling, serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and alkaline phosphatase (ALP) were significantly elevated (p<0.05), indicating hepatocellular damage; - Inflammatory Factors: Serum TNF-α and IL-6 levels were significantly upregulated (p<0.05), reflecting liver inflammation activation; 2. Pathological Indicators: - Liver Injury/Precancerous Lesions (3-6 weeks): HE staining showed destruction of liver lobular structure, sinusoidal dilation, periportal fibrosis, and inflammatory cell infiltration; hepatocellular adenoma and peliosis hepatis were visible at 6 weeks; - Liver Tumors (6-12 months): Typical pathological features of hepatocellular carcinoma were formed, with tumor tissue arranged in nests or cords and significant cellular atypia; 3. Immunohistochemical Indicators: - Proliferation Markers: Ki-67 (a cell proliferation marker) was strongly positively expressed in hepatocytes (dark brown staining); Hepatocyte-specific markers: Strong positive expression of HSA (hepatocyte-specific antigen) in the cytoplasm (golden yellow granules) indicates abnormal hepatocyte proliferation.

*Precautions： After being kept alive for 14 days, i.e., on day 15, the mice were sacrificed to measure serum biochemical (liver and kidney) parameters according to standard methods.

*References：Akree LS,et,al. In silico and in vivo hepatoprotective activity of the synthesized 5-benzylidene-2-thiohydantoin against diethylnitrosamine-induced liver injury in a rat model. Sci Rep. 2023 Mar 22;13(1):4681.