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Updated: Jun 1, 2026
Reprograming Model of Human Monocyte-derived Macrophages for In-vitro Assays
Published on: April 18, 2025
[Andrographolide inhibits extracellular signal-regulated kinase 1/2 signaling pathway in activated macrophages]
Lin-hua Qin1, Jiao Lü, Lin Kong
1Department of Digestive Diseases, the 411 Hospital of People's Liberation Army, Shanghai 200081, China. ando_8171@yahoo.com.cn
Objective:
To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages.
Methods:
LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay.
Results:
Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels.
Conclusion:
In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.
Insights
Andrographolide inhibits tumor necrosis factor-α (TNF-α) by suppressing the extracellular signal-regulated kinase 1/2 (ERK1/2) pathway in activated macrophages. This natural compound demonstrates potential therapeutic applications without cytotoxicity.
Area of Science:
- Immunology
- Pharmacology
- Molecular Biology
Context:
- Macrophages play a crucial role in inflammatory responses.
- Lipopolysaccharide (LPS) is a potent activator of macrophages, inducing pro-inflammatory cytokine production.
- The extracellular signal-regulated kinase 1/2 (ERK1/2) pathway is a key signaling cascade involved in macrophage activation and cytokine expression.
Purpose:
- To elucidate the effects of andrographolide on the ERK1/2 signaling pathway.
- To investigate andrographolide's impact on tumor necrosis factor-α (TNF-α) expression in LPS-activated macrophages.
- To determine the mechanism by which andrographolide modulates inflammatory responses.
Summary:
- Andrographolide, at concentrations up to 100 μg/mL, exhibited no cytotoxicity in LPS-activated mouse peritoneal macrophages.
- The study found that andrographolide significantly inhibited ERK1/2 phosphorylation in a dose-dependent manner.
- Andrographolide suppressed both mRNA and protein levels of TNF-α, similar to the effect of PD98059, an ERK1/2 pathway inhibitor.
Impact:
- Andrographolide demonstrates potential as an anti-inflammatory agent by targeting the ERK1/2 pathway.
- These findings suggest andrographolide could be a therapeutic candidate for inflammatory diseases driven by TNF-α.
- The research provides mechanistic insights into andrographolide's immunomodulatory properties.
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