Macrophage KLF15 prevents foam cell formation and atherosclerosis via transcriptional suppression of OLR-1.

BACKGROUND: Macrophage-derived foam cells are a hallmark of atherosclerosis. Scavenger receptors, including lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (OLR-1), are the principal receptors responsible for the uptake and modification of LDL, facilitating macrophage lipid load and the uptake of oxidized LDL by arterial wall cells. Krüppel-like factor 15 (KLF15) is a transcription factor that regulates the expression of genes by binding to the promoter during transcription. Therefore, this study aimed to investigate the precise role of macrophage KLF15 in atherogenesis. METHODS: We used two murine models of atherosclerosis: mice injected with an adeno-associated virus (AAV) encoding the Asp374-to-Tyr mutant version of human PCSK9, followed by 12 weeks on a high-fat diet (HFD), and ApoE-/-- mice on a HFD. We subsequently injected mice with AAV-KLF15 and AAV-LacZ to assess the role of KLF15 in the development of atherosclerosis in vivo. Oil Red O, H&E, and Masson's trichome staining were used to evaluate atherosclerotic lesions. Western blots and RT-qPCR were used to assess protein and mRNA levels, respectively. RESULTS: We determined that KLF15 expression was downregulated during atherosclerosis formation, and KLF15 overexpression prevented atherosclerosis progression. KLF15 expression levels did not affect body weight or serum lipid levels in mice. However, KLF15 overexpression in macrophages prevented foam cell formation by reducing OLR-1-meditated lipid uptake. KLF15 directly targeted and transcriptionally downregulated OLR-1 levels. Restoration of OLR-1 reversed the beneficial effects of KLF15 in atherosclerosis. CONCLUSION: Macrophage KLF15 transcriptionally downregulated OLR-1 expression to reduce lipid uptake, thereby preventing foam cell formation and atherosclerosis. Thus, our results suggest that KLF15 is a potential therapeutic target for atherosclerosis.

Phosphatidylethanolamine aggravates Angiotensin II-induced atrial fibrosis by triggering ferroptosis in mice.

As atrial fibrosis is the main feature of atrial structural remodeling, inhibiting atrial fibrosis is crucial to the prevention of atrial fibrillation (AF) progression. Research has shown the correlation between abnormal lipid metabolism and AF progression. However, the effect of specific lipids on atrial fibrosis remains unclear. In the present study, we applied ultra-high-performance lipidomics to analyze the lipid profiles in patients with AF and identify phosphatidylethanolamine (PE) as the differential lipid associated with AF. To detect the effect of the differential lipid on atrial fibrosis, we performed the intraperitoneal injection of Angiotensin II (Ang II) to mice to induce atrial fibrosis and supplemented PE in diets. We also treated atrial cells with PE to evaluate the cellular effect of PE. We found that PE supplementation aggravated atrial fibrosis and increased the expression of the fibrosis-related protein in vitro and in vivo. Moreover, we detected the effect of PE on the atrium. We found that PE increased oxidation products and regulated the expression of ferroptosis-related proteins, which could be alleviated by a ferroptosis inhibitor. PE increased peroxidation and mitochondrial damage in vitro, which promoted cardiomyocyte death induced by Ang II. Examination of protein expression in cardiomyocytes indicated that PE triggered ferroptosis and caused cell death to participate in myocardium fibrosis. In summary, our findings demonstrated the differential lipid profiles of AF patients and revealed the potential effect of PE on atrial remodelling, suggesting that inhibition of PE and ferroptosis might serve as a potential therapy to prevent AF progression.

Normothermic ex vivo heart perfusion with NLRP3 inflammasome inhibitor Mcc950 treatment improves cardiac function of circulatory death hearts after transplantation.

Background: The utilization of donation after circulatory death (DCD) hearts can enlarge the donor pool. However, DCD hearts suffer from serious ischemia/reperfusion injury (IRI). Recent studies found that the activation of NLRP3 inflammasome could play a significant role in organ IRI. Mcc950, which is a novel inhibitor of the NLRP3 inflammasome, can be applied to treat various kinds of cardiovascular diseases. Therefore, we hypothesized that the treatment of mcc950 could protect DCD hearts preserved with normothermic ex vivo heart perfusion (EVHP) against myocardial IRI via inhibiting NLRP3 inflammasome in a rat heart transplantation model of DCD. Methods: Donor-heart rats were randomly divided into four groups: Control group; Vehicle group; MP-mcc950 group; and MP + PO-mcc950 group. Mcc950 was added into the perfusate of normothermic EVHP in the MP-mcc950 and MP + PO-mcc950 groups, and was injected into the left external jugular vein after heart transplantation in the MP + PO-mcc950 group. Cardiac functional assessment was performed. The level of oxidative stress, inflammatory response, apoptosis, and NLRP3 inflammasome-associated protein of donor hearts were evaluated. Results: The treatment with mcc950 significantly increased the developed pressure (DP), dP/dtmax, and dP/dtmin of the left ventricular of DCD hearts at 90 min after heart transplantation in both MP-mcc950 and MP + PO-mcc950 groups. Furthermore, mcc950 added into perfusate and injected after transplantation in both MP-mcc950 and MP + PO-mcc950 groups significantly attenuated the level of oxidative stress, inflammatory response, apoptosis, and NLRP3 inflammasome compared with the vehicle group. Conclusions: Normothermic EVHP combined with mcc950 treatment can be a promising and novel DCD heart preservation strategy, which can alleviate myocardial IRI via inhibiting NLRP3 inflammasome.

Normothermic Ex Vivo Heart Perfusion with Mesenchymal Stem Cell-Derived Conditioned Medium Improves Myocardial Tissue Protection in Rat Donation after Circulatory Death Hearts.

Objective: Adopting hearts from donation after circulatory death (DCD) is a promising approach to enlarge the donor pool. Nevertheless, DCD hearts experience severe warm ischemia/reperfusion (I/R) injury. Recent studies have demonstrated that conditioned medium (CM) derived from bone marrow mesenchymal stem cells (BMSCs) has the potential of reducing organ I/R injury. Therefore, we investigated whether DCD heart preservation with normothermic ex vivo heart perfusion (EVHP) and BMSCs-CM treatment could alleviate myocardial warm I/R injury in the DCD hearts. Methods: We randomly divided donor rats into two groups: (1) DCD-Control group and (2) DCD-CM group. Before DCD heart preservation with the normothermic EVHP system for 105 minutes, rats suffered from a 25-minute warm ischemia injury in the DCD procedure. Vehicle or CM (300 µl) was added to the perfusate at the beginning of the perfusion process. The cardiac function of DCD hearts in the DCD-Control and DCD-CM groups was measured every 30 minutes. Besides, non-DCD hearts were harvested from the beating-heart rats. Results: The antibody array demonstrated that the CM contained 14 bioactive factors involved in apoptosis, inflammation, and oxidative stress. Warm ischemia injury resulted in a significant increase in the level of oxidative stress, inflammation, and apoptosis in the DCD hearts of DCD-Control group. Furthermore, compared with the DCD-Control group, CM treatment increased the developed pressure, dP/dtmax and dP/dtmin of the left ventricular in the DCD hearts during a 90-minute EVHP. Moreover, the administration of CM attenuated the level of oxidative stress, inflammation, and apoptosis in the DCD hearts of the DCD-CM group. Conclusions: Normothermic EVHP combined with CM treatment can alleviate warm I/R injury in the DCD hearts by decreasing the level of oxidative stress, inflammatory response, and apoptosis, which might alleviate the shortage of donor hearts by adopting DCD hearts.

Characterization of the complete chloroplast genome of Quercus virginiana Mill. (Fagaceae).

The complete chloroplast genome of Quercus virginiana was sequenced with Illumina HiSeq 2000 platform. It was a typical quadruple structure as other plants of Quercus with 161,221 bp in length, including a large single-copy (LSC: 90,553 bp) region and a small single-copy (SSC: 19,016 bp) which were separated by a pair of inverted repeats (IRa, b: 25,826 bp) region. The overall GC content is 36.9%. A total of 131 genes was annotated which contained 86 protein-coding genes including the Trans splicing gene of rps12, 37 tRNA genes, and 8 rRNA genes. ML phylogenetic analysis compared with 17 expressed chloroplast genomes revealed that Q. virginiana was a sister to other species of Quercus, which were grouped together with five species of Section Quercus and another 12 species of Quercus were divided into another group.

[Comparison of Soil Bacterial Community Structure Between Paddy Fields and Dry Land in the Huixian Karst Wetland, China].

In order to explore the effect of land-use change on soil bacteria in wetland systems, the topsoil (0-20 cm) of a natural wetland (NW), paddy field (PF), and dry land (DL) were collected in the Huixian karst wetland. The α-diversity, species composition, and abundance of soil bacterial communities were analyzed using high-throughput sequencing. The effect of environmental factors on bacterial community structure was also examined. The results showed that the soil bacteria in the Huixian karst wetland can be divided into 49 phyla and 145 classes. The Shannon index of bacteria in the PF was significantly higher, and the Simpson index of bacteria in the NW is significantly lower, than in the other two land-use types. The dominant phyla (operational taxonomic units, OTUs>1%) in the NW were Proteobacteria (52.15%), Actinobacteria (15.16%), and Acidobacteria (8.80%); the dominant phyla in the PF were Proteobacteria (45.79%), Acidobacteria (17.20%), and Chloroflexi (11.75%); the dominant phyla in the DL were Proteus (51.42%), Acidobacteria (15.51%), and Chloroflexi (7.43%). The dominant classes (OTUs>1%) in the NW were α-Proteobacteria (17.98%), ß-Proteobacteria (13.72%), and Actinobacteria (13.13%); the dominant classes in the PF were Acidobacteria (14.35%), ß-Proteobacteria (13.37%), and δ-Proteobacteria (12.02%); the dominant classes in the DL were α-Proteobacteria (19.44%), Formobacteria (13.30%), and Acidobacteria (13.03%). Among the dominant OTUs (>0.3%), the dominant genera of in the NW were Sphingomonas (OTU2, 59), Micromonospora (OTU5, 24 and 50487), Gemmatimonas (OTU1), and Tenotrophomonas (OTU8); the dominant genera in the PF were Lysobacter (OTU4 and 115) and Aquabacterium (OTU33); the dominant genera in the DL were Sphingomonas (OTU85, 157 and 2916), Rhodanobacter (OTU19 and 52), and Penlobacterium (OTU60). A heatmap showed that there were significant differences in soil bacterial community structure among the three land-use types. Redundancy analysis showed that pH, soil organic carbon (SOC), total nitrogen (TN), alkali-hydrolyzable nitrogen (AN), exchangeable Mg2+, exchangeable Ca2+, soluble organic carbon (DOC), and available phosphorus (AP) were the main factors that affected the bacterial community structure in the Huixian karst wetland. These results indicate that changes in land-use types have significantly shaped the structure of soil bacterial communities in this area.