Work

Interleukin‐35 Ameliorates Obesity‐Associated Impairment of Endothelium‐Dependent Vasodilation via the STAT3‐MAPK/ERK‐eNOS Signaling Axis

Year: 2026

Type: article

Abstract: BACKGROUND: Obesity impairs endothelial nitric oxide synthase (eNOS)-nitric oxide (NO) signaling, triggering chronic inflammation, endothelial dysfunction and defective vasodilation, which drives cardiometabolic diseases. Existing metabolic therapies provide only indirect vascular protection and fail to resolve residual inflammatory-mediated cardiovascular risk. As an anti-inflammatory interleukin 12 (IL-12) family cytokine, interleukin 35 (IL-35) is suppressed in obesity and exerts vascular protective effects under inflammatory conditions, yet its function and underlying mechanisms in obesity‑related endothelial vasodilator dysfunction remain undefined. MATERIALS AND METHODS: An obese mouse model was established by 8-week high-fat diet (HFD) feeding, followed by 4-week intraperitoneal administration of recombinant mouse IL-35. Aortic-ring assays evaluated vasodilatory function. Palmitic acid (PA)-treated human umbilical vein endothelial cells (HUVECs), recapitulate lipotoxic endothelial injury, were treated with recombinant human IL-35. Western blotting, immunofluorescence, 4-amino-5-methylamino-2',7'-difluorofluorescein diacetate (DAF-FM DA) detection and immunoprecipitation-mass spectrometry (IP-MS) were applied to detect IL-35-subunit (Epstein-Barr virus-induced gene 3 [EBI3] and interleukin-12 subunit alpha [IL-12A]) levels, eNOS Ser1177 phosphorylation, phosphorylation of mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling molecules (cellular rapidly accelerated fibrosarcoma [cRAF], mitogen-activated protein kinase kinase [MEK], and ERK) and signal transducer and activator of transcription 3 (STAT3) activation, NO production, and eNOS-binding partners. Rescue assays were performed using Raf or ERK specific inhibitors and STAT3 small interfering RNA (siRNA). RESULTS: HFD mice displayed reduced aortic endothelial EBI3 expression, impaired endothelium-dependent vasorelaxation, decreased eNOS Ser1177 phosphorylation and lowered NO production; all these phenotypes were rescued by exogenous IL-35. IP-MS and immunoblotting demonstrated that IL-35 strengthened eNOS-ERK1 interaction and restored PA-repressed phosphorylation of cRAF, MEK, ERK and eNOS Ser1177, which were abrogated by Raf or ERK blockade. Furthermore, IL-35 recovered STAT3 phosphorylation suppressed by PA, while STAT3 knockdown blunted IL-35-driven MAPK/ERK activation, eNOS phosphorylation and subsequent NO production. CONCLUSION: IL-35 alleviates obesity-induced impairment of endothelium-dependent vasodilation via the STAT3-MAPK/ERK-eNOS axis, which elevates eNOS Ser1177 phosphorylation and NO synthesis. This work supports IL-35 as a prospective therapeutic candidate for obesity-associated vascular complications.

Source: Immunity Inflammation and Disease

DOI: 10.1002/iid3.70564

Authors: Kai Li, Jie Feng, Ren Gong, Yunman Liu, Yanqing Wu

Institutions: Nanchang University, Second Affiliated Hospital of Nanchang University, Second Hospital of Nanchang

Language: English

Topic: Nitric Oxide and Endothelin Effects

Subfield: Physiology (medical)

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $3,300