Vascular calcification (VC) plays as a critical role on cardiovascular disease (CVD) and acts as a notable risk factor in cardiovascular system. Vascular smooth muscle cells (VSMCs) calcification can be triggered by high phosphate treatment; however, the explicit mechanism remains unclear. In the present study, we isolated VSMCs from primary rat artery, applied β‐GP (β‐glycerophosphate) for inducing VSMCs calcification in vitro to explore the mechanism of phosphate‐induced calcification in VSMCs. Alizarin red staining was performed to assess the mineralization in VSMCs. Calcium deposition experiment was taken to evaluate the calcium content. ALP staining was determined to assess the ALP activity. The recombinant adenoviruses were constructed for the overexpression of Klotho and FGF23, respectively. qRT‐PCR and western blot analysis were subjected to measure the expression of Klotho/FGF23 and correlated genes among Wnt7b/β‐catenin pathway. We found that the calcium content was obviously increased and Alizarin red staining was positive in calcification group exposure with high phosphate in a time‐dependent manner. The expression of Klotho and FGF23 was significantly decreased in the calcification group. However, overexpression of Klotho and FGF23 markedly reversed VSMCs calcification stimulating with high phosphate treatment. Moreover, Wnt7b/β‐catenin inhibitor DKK1 could partly attenuate the effect of high phosphate on calcified VSMCs. These findings demonstrated that Klotho/FGF23 axis could modulate high phosphate‐induced VSMCs calcification via Wnt7b/β‐catenin signaling pathway. Our findings unravel that Klotho/FGF23‐ Wnt7b/β‐catenin axis functions as a crucial role in the VSMCs calcification.

Thoracic radiotherapy patients have higher risks of developing radiation-induced heart disease (RIHD). Ionizing radiation generates excessive reactive oxygens species (ROS) causing oxidative stress, while Momordica. charantia and its extract have antioxidant activity. Plant-derived extracellular vesicles (EVs) is emerging as novel therapeutic agent. Therefore, we explored the protective effects of Momordica. charantia-derived EVs-like nanovesicles (MCELNs) against RIHD. Using density gradient centrifugation, we successfully isolated MCELNs with similar shape, size, and markers as EVs. Confocal imaging revealed that rat cardiomyocytes H9C2 cells internalized PKH67 labeled MCELNs time-dependently. In vitro assay identified that MCELNs promoted cell proliferation, suppressed cell apoptosis, and alleviated the DNA damage in irradiated (16 Gy, X-ray) H9C2 cells. Moreover, elevated mitochondria ROS in irradiated H9C2 cells were scavenged by MCELNs, protecting mitochondria function with re-balanced mitochondria membrane potential. Furthermore, the phosphorylation of ROS-related proteins was recovered with increased ratios of p-AKT/AKT and p-ERK/ERK in MCELNs treated irradiated H9C2 cells. Last, intraperitoneal administration of MCELNs mitigated myocardial injury and fibrosis in a thoracic radiation mice model. Our data demonstrated the potential protective effects of MCELNs against RIHD. The MCELNs shed light on preventive regime development for radiation-related toxicity.

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In this work, our primary objective was to examine the interrelationship among the serum level of chemokine (C-C motif) ligand 2 (CCL2) and plaque characteristics in coronary culprit lesions. The clinical data of 116 coronary heart disease patients who were hospitalized in the Department of Cardiology of Henan Province People's Hospital from February 2015 to June 2017 were retrospectively analyzed. The study population was subdivided according to the concentration of CCL2 into low CCL2 group and high CCL2 group. The levels of blood lipid, creatinine, and uric acid were measured, and patients underwent coronary angiography. The characteristics of the culprit lesions were detected by intravascular ultrasound, and the correlation between the serum markers and the characteristics of coronary artery plaque was analyzed. Moreover, the coronary artery disease dataset from the Gene Expression Omnibus database was downloaded and the genes regulated were analyzed by CCL2 using gene set enrichment analysis (GSEA). Patients with high CCL2 group had higher LDL-C level and L/H ratio, and lower HDL-C level than the low CCL2 group. Compared with low-level CCL2 group, coronary plaque in the high CCL2 group had higher eccentric plaque and plaque rupture, and thin cap fibroatheromas, fibrofatty and necrotic core and lower fibrous tissue. CCL2 was positively correlated with the percentage of fibrofatty and necrotic core, and negatively correlated with the percentage of fibrous tissue. Furthermore, GSEA analysis showed that samples with high CCL2 expression were enriched for genes involved in different pathways, such as cell adhesion molecules and Nod-like receptor signaling pathway. The CCL2 level was correlated with vulnerable plaques of coronary artery and had certain value in detecting vulnerable plaques. These results indicated that CCL2 could be regarded as a clinical prognostic biomarker for coronary artery disease. Impact statement Vulnerable plaques are plaques which are susceptible to rupture or thrombosis and trigger a series of adverse events such as coronary disorders. CCL2 is a soluble basic protein belonging to the CC subfamily. Previous studies have been investigated on the correlation between inflammatory factors and clinical events, but there are few studies on the correlation between CCL2 and plaque characteristics. Our study found that the high expression of CCL2 is involved in multiple processes in the genesis and progression of coronary artery disease, and would be a potential clinical prognostic indicator. In addition, high expression of CCL2 may be related to gene pathways such as Nod-like receptor signaling pathway, suggesting that CCL2 is involved in the inflammatory response and immune process of coronary artery disease.