Human Hepatic Stellate Cell Oxidative Stress Model

Human Hepatic Stellate Cell Oxidative Stress Model Figure 1. Oxidative stress model of activated human hepatic stellate cell. (Andueza et al., 2018)

Hepatic stellate cell activation is the central link of the occurrence and development of hepatic fibrosis. Therefore, it is very necessary to construct a scientific hepatic stellate cell oxidative stress model in vitro.

Creative Bioarray has constructed the oxidative stress model of activated human hepatic stellate cell line (LX-2) through H₂O₂ and Fe-NTA induction. This model can basically represent one of the main oxidative stress molecules in the liver. It will provide an effective tool for testing the specific role of drugs in oxidative stress of liver fibrosis and ensure the reliability of the research results.

Model Information

Model application

Human hepatic stellate cell oxidative stress model can be used for drug administration experiment, pharmacopharmacology, pharmacodynamic evaluation and pharmacokinetic analysis to help customers study liver cancer, hepatitis, liver fibrosis and other diseases and seek new drugs for prevention and treatment.

H₂O₂ Modeling method of LX-2

H₂O₂ was used to simulate the actual oxidative stress environment in the liver. H₂O₂ mainly simulates the H₂O₂ reactive oxygen intermediates widely existing in the liver. After H₂O₂ treatment, the superoxide in cells will increase significantly, which will stimulate the oxidative stress response of cells.

Fe-NTA Modeling method of LX-2

The cells were isolated and cultured according to standard procedures. All cells were was inoculated on 96 well plates, in which LX-2 cells were cultured for 5 days after separation. After overnight culture, a certain concentration of Fe-NTA was added.

H₂O₂ Modeling Vs. Fe-NTA Modeling

H₂O₂ mainly simulates the H₂O₂ reactive oxygen intermediates widely existing in the liver. The Fe overload model caused by Fe-NTA is mainly manifested in lipid peroxidation, so it is milder than H₂O₂. The iron overload model is mainly used to trigger lipid peroxidation in the liver. The synergistic reaction between iron and arachidonic acid mainly occurs in the process of liver fibrosis induced by alcoholic fatty liver.

SOP for Modeling Effect Guarantee

Intracellular superoxide detection

After the cells were treated accordingly, the superoxide anion fluorescent probe working solution was added, incubated at 37 ℃ for 60min, washed twice with s buffer, and observed by green excitation under fluorescence microscope.

MTT assay

After reaching the set time, continue to culture MTT reagent in the sample for 4h, terminate the culture, suck and discard the culture medium, add DMSO, shake to fully dissolve the crystal, measure the OD value at 492 nm of the microplate reader, and use 630nm as the reference wavelength.

Other Services

In addition to the human hepatic stellate cell oxidative stress model, Creative Bioarray can establish a variety of cellular oxidative stress and other stress models (nervous system, reproductive system, skin system, reproductive system, etc.) according to the specific needs of customers. Colleagues can provide administration experiments, pharmacopharmacology, efficacy evaluation and pharmacokinetic analysis, and assist customers in all-round scientific research projects.

Creative Bioarray is dedicated to providing high-quality products, comprehensive services, and tailored solutions to support and facilitate life sciences and pharmaceutical research and development. If you have any questions or needs, please contact us, and our customer service staff will help you at the first time.

Reference

Andueza A, Garde N, García-Garzón A, Ansorena E, López-Zabalza MJ, Iraburu MJ, Zalba G, Martínez-Irujo JJ. NADPH oxidase 5 promotes proliferation and fibrosis in human hepatic stellate cells. Free Radic Biol Med. 2018 Oct;126:15-26.

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