Related Experiment Video
Updated: Mar 24, 2026
Author Spotlight: High-Throughput Measurement of Intracellular ROS Levels in Hepatocellular Lines
Published on: January 19, 2024
Global protein expression dataset acquired during isoniazid-induced cytoprotection against H2O2 challenge in HL-60
Saifur R Khan1, Argishti Baghdasarian1, Richard P Fahlman2
1Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, Canada.
Abstract:
Isoniazid (INH) is one of the first-line anti-tuberculosis drugs. Its effect on oxidative stress, however, is unknown. Here we used a model of oxidative stress by employing glucose/glucose oxidase (GOx), which (based on the availability of glucose and oxygen) is known to produce H2O2. This reaction induces oxidative stress culminating in necrotic cell death in HL-60 cells (a human promyelocytic leukemia cell line). The changes in protein levels have been quantified using global proteome expression changes through stable isotope labeling by amino acids in cell culture (SILAC) followed by LC-MS/MS analysis. A total of 1459 and 1712 proteins were identified in forward and reverse experiments, respectively. However, only 390 proteins were reproducibly identified in both samples. These 390 proteins were taken into account for further analysis which has been described in "Cytoprotective effect of isoniazid against H2O2 derived injury in HL-60 cells" [1].
Insights
Isoniazid (INH), a tuberculosis drug, was investigated for its effects on oxidative stress. This study found that INH protects HL-60 cells from hydrogen peroxide (H2O2)-induced necrotic cell death.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Tuberculosis remains a significant global health challenge, necessitating effective therapeutic strategies.
- Isoniazid (INH) is a cornerstone first-line anti-tuberculosis medication.
- The impact of INH on cellular oxidative stress pathways remains largely unexplored.
Purpose of the Study:
- To investigate the potential cytoprotective effects of Isoniazid (INH) against oxidative stress.
- To elucidate the mechanisms underlying INH's action in a cellular model of oxidative damage.
Main Methods:
- Establishment of an oxidative stress model in HL-60 cells using glucose/glucose oxidase (GOx) to generate hydrogen peroxide (H2O2).
- Quantitative proteomic analysis using stable isotope labeling by amino acids in cell culture (SILAC) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS).
- Identification and quantification of protein level changes in response to H2O2-induced stress and INH treatment.
Main Results:
- A total of 390 proteins were reproducibly identified across both forward and reverse proteomic experiments.
- The study identified key protein alterations associated with oxidative stress and potential INH-mediated protection.
- Detailed proteomic data provided insights into cellular responses to H2O2 and the influence of INH.
Conclusions:
- Isoniazid (INH) demonstrates a cytoprotective effect against hydrogen peroxide (H2O2)-induced necrotic cell death in HL-60 cells.
- The findings suggest that INH may modulate cellular pathways involved in mitigating oxidative stress.
- Further research is warranted to fully understand INH's role in combating oxidative damage in tuberculosis treatment.
