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Animal models of hepatic fibrosis/cirrhosis

Animal models of hepatic fibrosis/cirrhosis

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Liver fibrosis/liver cirrhosis
Psychiatric & neurological disease models

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Model Introduction

Model Introduction

Platform Service Advantages

Platform Service Advantages

Experimental results

Experimental results

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Liver fibrosis/liver cirrhosis

Introduction to Animal Models of Liver Fibrosis

It is primarily a pathological concept referring to the excessive proliferation and abnormal deposition of extracellular matrix (ECM) components within hepatic tissue, leading to structural and/or functional abnormalities of the liver. At its core, this process represents the liver’s reparative response to chronic injury, analogous to scar formation in human skin following trauma.

Currently, both domestically and internationally, the commonly used methods for establishing hepatic fibrosis models include carbon tetrachloride (CCl₄)-induced, N‑nitrosodimethylamine (NDMA)-induced, bile duct ligation–induced, and diet‑induced models. However, the most classic and widely employed approach is the CCl₄‑induced hepatic fibrosis animal model.

Introduction to Animal Models of Liver Cirrhosis

CCl₄, as the earliest and most widely used inducer of hepatic fibrosis, induces chronic liver injury by directly damaging hepatocytes, disrupting their cellular components, and leading to steatosis and necrosis. Ethanol, acting as an enzyme inducer, impairs mitochondria, increases hepatic oxygen consumption, and can also enhance CCl₄… 4 Regarding hepatotoxicity, the simultaneous action of both agents can promote the development of hepatic cirrhosis.

Therefore, the CCl₄ model is frequently employed in experimental studies to investigate the pathogenesis of drug‑induced liver cirrhosis and to evaluate the efficacy of anti‑cirrhotic agents. The CCl₄‑induced hepatic cirrhosis animal model is characterized by its simplicity of operation, low cost of materials, and typical pathological features, and it can be established via oral administration, intraperitoneal injection, or subcutaneous injection.

Service Advantages of the Aniphe Biolaboratory Liver Fibrosis/Liver Cirrhosis Animal Model Platform

Our platform boasts mature and well‑established techniques for establishing animal models of hepatic fibrosis and cirrhosis, capable of meeting the diverse needs of scientific research and new drug development.

Comprehensive model coverage

The model has a high degree of simulation accuracy.

The animal model progressively develops hepatocellular injury, elevated liver enzymes, hepatic fibrosis, and nodular hyperplasia, which may advance to typical cirrhotic changes, mirroring the clinical disease course.
Standardization of the modeling process

The model exhibits stable traits.

The modeling success rate is high, individual variability within groups is minimal, and experimental reproducibility is excellent, ensuring the reliability of the data.
Multidimensional Evaluation System

Standardized Feeding and Management

All animals were housed under SPF‑grade conditions, with their health status and various physiological parameters monitored at scheduled intervals, ensuring full traceability of experimental data.
Adapted for multi-cycle studies

Comprehensive testing items

It encompasses multidimensional assessment and evaluation, including hepatic biochemical markers, liver histopathology, fibrosis‑related factors, and imaging studies.
Standardization of Data Delivery

End-to-end one-stop service

We undertake the full spectrum of services, including model generation, animal care, sample collection, and biomarker analysis, thereby supporting both basic research and preclinical drug development for regulatory submissions.

Animal model of liver fibrosis

Selection of experimental animals: Because female rats readily develop tolerance to CCl₄, resulting in a low success rate for model establishment, CCl₄–induced hepatic fibrosis models typically employ mice or male rats as the animal subjects, with the primary routes of exposure being gavage, intraperitoneal injection, or subcutaneous injection.

Rat model preparation method:

01 Prepare a CCl₄ solution by mixing CCl₄ and olive oil at a volume ratio of 1:4, and set aside.

02 CCl₄ was administered by intraperitoneal injection according to the body weight of the rats, twice weekly for 6 weeks. The blank control group received olive oil only.

03 After two consecutive weeks of CCl₄ administration, three rats each from the control group and the model‑induced group were randomly selected for assessment of ALT, AST, total bilirubin (TBL), and histological examination to confirm successful model establishment.

04 If the model is successful, drug evaluation can proceed; after an additional 4-week treatment period, the rats can be euthanized, and liver fibrosis–related biomarkers can be assessed to evaluate the drug’s efficacy.

Experimental results

HE staining assay

HE staining was used to assess the liver fibrosis model: in normal liver tissue, hepatocytes exhibited abundant, reddish cytoplasm with no obvious steatosis; hepatocytes were arranged radially around the central vein.

In the model group, hepatocytes exhibited marked steatosis, with some showing hydropic degeneration; the lobular architecture was indistinct, and fibrous tissue was markedly proliferated, forming fibrous septa that partitioned the hepatic lobules. Scattered inflammatory cell infiltration was observed within these fibrous septa.

 

 

Pathological sections of hepatic fibrosis in rats

Observation of Gross Specimens of Rat Liver

In the normal group, the liver surface was bright red, smooth and soft, without nodules or granules; the hepatic margins were sharp, and both the shape and size were within normal limits.

In the liver fibrosis model group, the liver surface appeared darker than that of normal tissue, with a slightly firmer consistency and a granular texture. The hepatic margins were blunter compared to the control group, and the capsule was swollen.

 

 

Mouse model induction methods

01 Adult male mice were administered a CCl₄–olive oil solution by subcutaneous injection every 5 days for 3 consecutive months. Mice in the control group received an equivalent volume of olive oil vehicle via intraperitoneal injection.

02 Animals are fed a standard diet, and their activity, mental state, and eating behavior are observed; body weight is recorded weekly.

03 Prior to the end of the experiment, mice were fasted for 6 hours, after which they were euthanized and their livers were harvested for histopathological examination and analysis of relevant proteins.

 

Precautions

At 30 days post‑modeling, a small amount of fibrous tissue proliferation was observed in the livers of half of the mice in model group 01.

At 60 days, most animals exhibited mild to moderate fibrous tissue hyperplasia.

At 90 days, all animals exhibited moderate to extensive fibrous tissue hyperplasia, and the hepatic proline content also increased progressively with prolonged modeling duration; it was elevated at 60 days and showed an even more pronounced increase by 90 days.

04 The hepatic malondialdehyde content and serum alanine aminotransferase levels were significantly elevated at all stages of model establishment, and by day 90, the plasma albumin ratio had markedly decreased.

 

Experimental results

Western blot results

Compared with the control group, the CCl₄‑treated group exhibited a significant upregulation of Collagen‑1 and α‑SMA protein expression, indicating that CCl₄ successfully induced hepatic fibrosis in mice and that the model was effectively established.

 

 

Expression of proteins associated with hepatic tissue fibrosis

Animal model of liver cirrhosis

Carbon tetrachloride (CCl₄), the earliest and most widely used inducer of hepatic fibrosis, elicits chronic liver injury by directly damaging hepatocytes, disrupting cellular components, and inducing steatosis and necrosis. Ethanol acts as an enzyme inducer, impairing mitochondrial function and increasing hepatic oxygen consumption; it also enhances the toxic effects of CCl₄ on hepatocytes. The combined action of these two agents promotes the development of liver cirrhosis.

Therefore, the CCl₄ model is frequently employed in experimental studies to investigate the pathogenesis of drug‑induced liver cirrhosis and to evaluate the therapeutic efficacy of anti‑cirrhotic agents. The CCl₄‑induced hepatic fibrosis animal model is characterized by its simplicity of operation, low cost of materials, and typical pathological features, and it can be established via oral administration, intraperitoneal injection, or subcutaneous injection.

CCl₄-Induced Animal Model of Liver Cirrhosis

Selection of experimental animals: Adult male rat

Modeling method:

The experiment established a hepatic fibrosis animal model using the CCl₄‑induced method, with intraperitoneal or subcutaneous injection of a CCl₄ oil solution as the core modeling approach. Animals were administered the agent repeatedly at predetermined intervals over several weeks. To accelerate or intensify disease progression, some protocols additionally employed a high‑fat, high‑cholesterol diet and ethanol‑supplemented drinking water. By combining chemical liver injury with metabolic stimulation, chronic inflammation, hepatocellular necrosis, and fibrous tissue proliferation were induced, successfully yielding a stable hepatic fibrosis model.

Changes in liver wet weight

In the CCl₄–induced model, the early stage was characterized by hepatic lobe distortion and adhesions. By week 12, as fibrosis progressed, the liver surface became granular, and the wet weight of the liver was significantly higher than that of the control group (P<0.05).

 

 

Figure A shows the gross appearance of the liver in the control group rats.

Figures B–D show gross photographs of the livers of rats at 4, 8, and 12 weeks after CCl₄–induced liver fibrosis, respectively.