Inducing Acute Liver Injury in Rats via Carbon Tetrachloride (CCl4) Exposure Through an Orogastric Tube
April 28th, 2020
This protocol describes a common and feasible method of inducing acute liver injury (ALI) via CCl4 exposure through an orogastric tube. CCl4 exposure induces ALI through the formation of reactive oxygen species during its biotransformation in the liver. This method is used to analyze the pathophysiology of ALI and examine different hepatoprotective strategies.
I would like to present our work about induced acute liver injury in rats via carbon tetrachloride exposure through orogastric tube. Select adult male Sprague-Dawley rats weighing 300 to 350 gram. Provide rat chow and water ad libitum.
Perform all experiment between six AM and 12 PM.The animal care comedy of the Ben-Gurion University of the Negev, Israel approved all treatment and testing procedure used in this protocol. Maintain rats at room temperature of 22 Celsius degree with 12 hour lights and 12 hour dark cycles. Cannulate the tail vein with a 22 gauge catheter.
Collect blood samples at baseline. Blood biochemical analysis include the measurement of serum GOT, GPT, and total bilirubin. Examination of liver enzymes and total bilirubin level were carried out in the biochemical laboratory of Soroka Medical Center.
Caution:exposure to high concentration of carbon tetrachloride, including vapor or skin absorption can affect the central nervous system, degenerate the liver and kidney, and prolonged exposure may lead to coma or death. Prepare a 50 percent solution of carbon tetrachloride mixed with olive oil as vehicle in one to one ratio. Induce the pathotoxicity in vivo by carbon tetrachloride administration via an orogastric tube.
Insert an orogastric tube through the oral cavity of the rat. Expose the rats to different doses of carbon tetrachloride. One milliliter per kilo for mild acute liver injury, two point five milliliter per kilo for moderate acute liver injury, or five milliliter per kilo for severe acute liver injury.
For sham group, expose the rats to five milliliter per kilo only olive oil. Collect blood samples at 24 hours from carbon tetrachloride exposure. Blood biochemical analysis include the measurement of serum GOT, GPT, and total bilirubin.
Place dead rat on dissecting board in dorsal recombinant with the abdominal facing upwards and shave her abdomen. Using scalpel, incise the full length of the ventrum from the anus to the chin. Reflect the skin.
With the scalpel, incise the abdominal wall from the anus to the xiphoid cartilage to expose the abdominal viscera. Transfer the liver into a petri dish. Perform a hematoxylin and eosin staining.
Examine the slices under microscope magnification of 200 times with a 20 millimeters objective length. The liver section should be graded by a specialized pathologist, blinded to the treatment protocol. The score of zero indicates no liver abnormalities, one to two indicates mild liver injury, three to four indicates moderate liver injury, and five to six indicates severe liver injury.
The various groups of rats at different times are shown on the scheme. 24 hours after inducing acute liver injury, there was a significant increase in GOT, GPT, and total bilirubin levels, depending on the carbon tetrachloride exposed as compared to the control group. P value was less than zero point zero zero one.
Histopathological changes were observed after 24 hours in all groups after carbon tetrachloride intoxication in various doses. Histological grading score at 24 hours between all three interventional groups to the control group was significantly higher. P value was less than zero point zero five.
Our research group presented the common and physical method of induced acute liver injury in rats via carbon tetrachloride exposure through orogastric tube. This method is used to investigate the pathophysiology of acute liver injury and to test different about the protective strategy. Thank you very much for your attention.
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Overview
This protocol describes a method for inducing acute liver injury (ALI) in rats using carbon tetrachloride (CCl4) exposure via an orogastric tube. The process leads to the formation of reactive oxygen species in the liver, allowing researchers to study ALI pathophysiology and hepatoprotective strategies.
Key Study Components
Area of Science
- Neuroscience
- Biology
- Hepatology
Background
- Acute liver injury (ALI) is a significant health concern.
- CCl4 is a well-known hepatotoxic agent.
- Understanding ALI mechanisms can aid in developing treatments.
- This study uses a rat model for experimental analysis.
Purpose of Study
- To induce ALI in a controlled environment.
- To analyze the effects of CCl4 on liver function.
- To evaluate potential hepatoprotective strategies.
Methods Used
- Adult male Sprague-Dawley rats were selected for the study.
- Rats were provided with chow and water ad libitum.
- Experiments were conducted during specific hours to maintain consistency.
- Tail vein cannulation was performed for substance administration.
Main Results
- Induction of ALI was successfully achieved using CCl4.
- Reactive oxygen species were measured in liver tissues.
- Different hepatoprotective strategies were evaluated.
- Results contribute to understanding ALI mechanisms.
Conclusions
- The CCl4 exposure method is effective for studying ALI.
- Findings support further research into hepatoprotection.
- This model can be used for future therapeutic studies.
Frequently Asked Questions
Executive Industry Relevance
Robust animal models of acute liver injury are essential for de-risking early-stage hepatoprotective drug discovery and validating mechanistic hypotheses in hepatic pathophysiology. The CCl4-induced rat model enables quantitative assessment of liver injury severity and supports translational continuity from discovery to preclinical evaluation. This reproducible system provides a foundation for portfolio triage and predictive confidence in hepatoprotective candidate selection.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables interrogation of hepatotoxic mechanisms and pathway involvement in acute liver injury.
- Supports biological de-risking and functional validation of hepatoprotective targets.
- Facilitates predictive confidence in target engagement and mechanistic relevance.
Screening & Assay Development
- Provides a validated in vivo system for quantitative measurement of liver injury biomarkers (GOT, GPT, bilirubin).
- Enables reproducible grading of histopathological liver damage for assay standardization.
- Supports screening of hepatoprotective compounds with scalable and controlled injury induction.
Translational & Preclinical Research
- Aligns with disease-relevant endpoints for translational biomarker assessment.
- Ensures continuity from mechanistic discovery to preclinical validation of therapeutic strategies.
- Supports risk-adjusted advancement decisions based on quantitative and histological outputs.
Pipeline & Workflow Integration
This model integrates into the discovery-to-preclinical continuum, enabling hypothesis testing, compound screening, and translational validation in hepatic injury research.
- Discovery Biology: Supports mechanistic hypothesis testing and pathway clarification in liver injury.
- Screening: Delivers reproducible, quantitative biomarker outputs for compound evaluation.
- Analytics: Provides statistical comparison of injury severity and treatment efficacy across groups.
- Translational Research: Aligns preclinical findings with clinically relevant liver injury markers.
- Enterprise Reuse: Offers a standardized, scalable platform for repeated use in hepatoprotective R&D programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in hepatoprotective research.
- Operational Value: Delivers standardized, reproducible, and scalable in vivo workflows.
- Strategic Value: Enables informed go/no-go decisions and capital-efficient portfolio management.
- Portfolio Impact: Supports risk-adjusted prioritization and advancement of liver-targeted therapeutics.
Implementation Considerations
- Requires expertise in rodent handling, orogastric dosing, and histopathological analysis.
- Needs access to biochemical and histological analytical infrastructure for biomarker and tissue assessment.
- Demands cross-team standardization of dosing, sampling, and scoring protocols.
- Adaptable to various dosing regimens and injury severities for model system flexibility.
- Safety precautions are essential due to CCl4 toxicity and handling risks.
Biopharma FAQs
Chapters in this video
0:03
Introduction
0:23
Preparing Rats for the Experimental Procedure
1:15
Determination of Serum GOT, GPT and Total Bilirubin Baseline Level
1:44
The Procedure for Induction of Acute Liver Injury in Rats
3:03
Determination of Serum GOT, GPT and Total Bilirubin Level after 24 Hours
3:23
Liver Collection for Histological Examination
4:03
Histological Examination on Rats at Time 24 Hours
4:49
Results
5:38
Conclusion
