Luteolin modulates macrophage phenotypic switching via the AMPK-PPARγ pathway to alleviate ulcerative colitis in mice

Abstract

Ethnopharmacological relevance

Lonicerae japonicae flos (LJF), the dried flower bud or newly bloomed flower of Lonicera japonica Thunb., is widely used in Traditional Chinese medicine (TCM), exhibiting anti-inflammatory and immune-enhancing properties. Luteolin (Lut) is a major bioactive component of LJF, demonstrating a regulatory role in immune disorders. However, the specific role of Lut in regulating macrophage-mediated intestinal inflammation and its underlying molecular mechanisms have not yet been fully explored.

Aim of the study: This study was designed to explore whether Lut alleviates Ulcerative colitis (UC) in mice and to elucidate its underlying mechanism in intestinal inflammation.

Materials and methods

Mice were administered Dextran sodium sulfate (DSS) for 7 d to establish a UC model, followed by oral administration of Lut (12.5, 25, and 50 mg/kg body weight). RNA-sequencing (RNA-Seq) was used to screen signaling pathways. RAW264.7 cells were cultured and treated with Lut (6.25, 12.5, and 25 μM) and lipopolysaccharide (LPS, 1 μg/mL) for 24 h. To examine the role of the AMP-activated protein kinase (AMPK)/Peroxisome proliferator-activated receptor γ (PPARγ) signaling pathway, the cells were treated with compound C (an AMPK inhibitor) and GW9662 (a PPARγ antagonist).

Results

Lut suppressed the inflammation of DSS-induced colitis in vivo, attenuated DSS-induced clinical man-ifestations, reversed colon length reduction, and reduced histological injury. Lut induced a shift in the macrophage phenotype from classical (M1) to alternative (M2) by suppressing M1 marker gene expression and enhancing M2 marker gene expression following DSS or LPS induction. RNA-seq revealed that PPARγ was involved in the regulation of macrophages by Lut. Furthermore, the polarization effect of Lut on macrophages was shown to be mediated through the AMPK-PPARγ signaling pathway.

Conclusion

These findings indicate that Lut effectively ameliorates UC in mice through the activation of the AMPK-PPARγ signaling pathway, leading to the inhibition of macrophage M1 polarization and promotion of M2 polarization. This study provides insight into future research on the utilization of Lut-rich TCM dietary supplements as a prophylactic treatment strategy in the prevention of UC.