Non-Lethal Sonodynamic Therapy Inhibits Atherosclerotic Plaque Progression in ApoE-/- Mice and Attenuates ox-LDL-mediated Macrophage Impairment by Inducing Heme Oxygenase-1

Abstract

Background: Previous studies from our group showed that low-intensity sonodynamic therapy (SDT) has protective effects on atherosclerosis (AS). However, because the intensity of ultrasound passing through tissue is attenuated, the consequences of very low-intensity SDT, referred to as non-lethal SDT (NL-SDT), on atherosclerotic plaques are unclear. The aim of this study was to determine whether NL-SDT affects atherosclerotic plaques and to elucidate the possible underlying mechanisms. Methods: An AS model was established using ApoE^-/- mice fed a western diet. En face Oil Red O staining was used to measure atherosclerotic plaque size. Hematoxylin and eosin staining and immunohistochemical staining were used to observe plaque morphology and assess the location of macrophages and heme oxygenase 1 (HO-1). HO-1 mRNA and protein levels in AS plaques were evaluated by real-time PCR and western blotting. Human THP-1 cells and mouse peritoneal macrophages were used in this study. Western blotting was used to investigate the expression of cellular proteins after NL-SDT. Macrophage apoptosis was evaluated by TUNEL assays and flow cytometry with Annexin V/PI double staining. Intracellular reactive oxygen species (ROS) and mitochondrial membrane potential (MMP) were measured with 2′-7′-dichlorofluorescein diacetate (DCFH-DA) and 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl benzimidazolyl carbocyanine iodide (JC-1) staining, respectively. Results: NL-SDT significantly inhibited AS progression and reduced the necrotic core area. NL-SDT induced HO-1 expression in lesional macrophages and in cultured macrophages. NL-SDT activated the protein kinase B (AKT) and extracellular signal-related protein kinase (ERK) pathways and the transcription factor NF-E2-related factor 2 (Nrf2).NL-SDT significantly reduced oxidized LDL (ox-LDL)-induced macrophage MMP collapse, ROS production and cell apoptosis. Zinc protoporphyrin (ZnPP), a HO-1-specific inhibitor, reversed the protective effects of NL-SDT. Conclusion: NL-SDT inhibits atherosclerotic plaque progression and increases plaque stability. In vitro, NL-SDT has a protective effect on ox-LDL-induced macrophage impairment via HO-1.

A fluorescently labeled undecapeptide derived from a protein in royal jelly of the honeybee—royalisin—for specific detection of oxidized low‐density lipoprotein

“…Recent evidence strongly suggests that the oxidized low‐density lipoprotein (ox‐LDL), which is abundantly present in atherosclerotic plaques and is associated with plaque instability, is more important in the pathogenesis and progression of atherosclerosis than native LDL . Oxidized LDL contributes to the atherosclerotic plaque formation and progression by several mechanisms, including the induction of endothelial cell activation and dysfunction, macrophage foam cell formation and apoptosis, and smooth muscle cell migration and proliferation . Accordingly, the plasma levels of ox‐LDL are known to be elevated in patients with atherosclerosis‐related cardiovascular diseases .…”

“…[4][5][6][7][8][9] Oxidized LDL contributes to the atherosclerotic plaque formation and progression by several mechanisms, including the induction of endothelial cell activation and dysfunction, macrophage foam cell formation and apoptosis, and smooth muscle cell migration and proliferation. [10][11][12][13] Accordingly, the plasma levels of ox-LDL are known to be elevated in patients with atherosclerosis-related cardiovascular diseases. 14 Therefore, the probes for specific detection of ox-LDL in atherosclerotic plaques and in plasma are expected to be useful for the identification, diagnosis, prevention, and treatment of atherosclerosis.…”

Methods of Estimation and the Role of Respiratory Burst in the Pathogenesis of Infectious and Inflammatory Diseases

“…Именно эта форма красителя и регистрируется в клетках при проведении анализа с помощью методов проточ-ной цитофлуоримет рии. Например, Wang Y. et al (2017) на основе анализа с DCFH-DA охарактеризовали состояние респираторного взрыва макрофагов при формировании атеросклеротической бляшки [47].…”

Rapamycin Alleviated the Symptoms of Fat Embolism Syndrome by Modulating Autophagy in a Rat Model

“…Oil red O staining was performed according to the method [31]. The entire lung of the rats were cut open and stained with Oil Red O for 30 min and then washed with ethanol 3 times.…”