Jian-pi Qu-tan Hua-yu Decoction improves oxidative stress-induced inflammation in endothelial cells in atherosclerotic ApoE-/- mice through the NOX1-ROS-ERK1/2 pathway

Abstract

Background: Atherosclerosis (AS) is a key mechanism in cardiovascular diseases and a major target for interventions. Jian-pi Qu-tan Hua-yu Decoction (JPQTHYD), a herbal formula, has been shown to alleviate AS.

Objective: This study aimed to evaluate the therapeutic effect of JPQTHYD on AS and explore its molecular mechanisms.

Materials and methods: In vivo, we established a mouse model through a high-fat diet for 16 weeks combined with a 4-week exhaustive swimming experiment. The body weight and food intake of the mice were measured every 4 weeks. At the end of the 16 weeks, the moisture content of the mice's feces was measured, the morphology of the thoracic aorta and myocardium was observed using HE staining, and lipid deposition in the aorta and myocardium was assessed using Oil Red O staining. The ultrastructure of myocardial tissue was observed via transmission electron microscopy. Levels of TC, TG, and LDL-C were measured using an automatic biochemical analyzer. ELISA was used to detect the levels of ROS, IL-6, IL-10, TNF-α, hs-CRP, VCAM-1, and ICAM-1. In vitro, we induced HUVECs injury using 700 nM of Ang II. Cell viability was assessed using the CCK-8 assay, while ROS levels were measured by a ROS detection kit. NOX1 gene suppression was achieved using a NOX1 inhibitor. Protein expression levels of NOX1, ERK1/2, P-ERK1/2, VCAM-1, and ICAM-1 were measured by Western blot both in vivo and in vitro.

Results: 1. JPQTHYD improved fecal water content and exercise capacity in ApoE-/- mice. 2. JPQTHYD reduced TC, TG, LDL-C levels, decreased arterial intimal thickness, and inhibited atherosclerotic plaque formation. 3. JPQTHYD decreased proinflammatory factors and adhesion molecules by inhibiting the NOX1-ROS-ERK1/2 pathway. 4. In vitro, JPQTHYD suppressed endothelial inflammation by reducing NOX1-ROS-ERK1/2 signaling.

Conclusion: JPQTHYD reduced blood lipids, inhibited oxidative stress-induced inflammation, and alleviated AS in ApoE-/- mice, likely through the NOX1-ROS-ERK1/2 pathway. This study offers a novel investigation into the mechanisms and regulatory pathways through which traditional Chinese medicine contributes to the prevention and treatment of atherosclerosis.

Keywords: Atherosclerosis; Jian-pi Qu-tan Hua-yu Decoction; Nicotinamide adenine dinucleotide phosphateoxidase (NOX); Oxidative stress;Reactive oxygen species (ROS).