Fibulin 1, targeted by microRNA-24-3p, promotes cell proliferation and migration in vascular smooth muscle cells, contributing to the development of atherosclerosis in APOE-/- mice.

Extracellular matrix (ECM) and vascular smooth muscle cells (VSMCs) are the main components of atherosclerosis (AS) plaque. VSMCs participate in plaque formation through phenotypic transformation. The complex interplay between ECM and VSMCs plays vital roles in the progression of AS throughout the disease. An in-depth investigation into the functions of ECM-related molecules in VSMC development might contribute to deciphering the complexity of AS pathogenesis. In this study, the roles and molecular mechanisms of the ECM-related molecule Fibulin-1 (FBLN1) in the development of AS and VSMCs were explored using RNA sequencing, bioinformatics analysis, and cell experiments. Furthermore, the expression of FBLN1, as determined by western blot analysis, immunohistochemistry, and real-time quantitative PCR, was significantly increased in AS vascular samples compared to normal vascular samples. Silencing the FBLN1 through AAV viral injection in mice revealed an improvement in AS. Functional analyses revealed that FBLN1 promoted VSMC proliferation, migration, and wound healing. Combined with RNA sequencing and TargetScan7.2 prediction data, 22 microRNAs (miRNAs) were found to have the potential for direct interaction with the FBLN1 3'UTR in VSMCs. Among these 22 miRNAs, it was demonstrated that microRNA-24-3p (miR-24-3p) could negatively regulate FBLN1 expression by directly binding to the FBLN1 3'UTR. Moreover, miR-24-3p inhibited cell proliferation, migration, and wound healing, and suppressed the expression of Ki67, matrix metalloproteinase-2 and -9 (MMP2/9) by targeting FBLN1 in VSMCs. Meanwhile, inhibition of FBLN1 expression could restrain VSMC phenotypic transformation. In conclusion, miR-24-3p inhibited VSMC proliferation and migration by targeting FBLN1. Additionally, multiple miRNAs with the potential to interact with the FBLN1 3'UTR were identified. These findings might deepen our understanding of ECM gene regulatory networks and the complex etiology of AS.

Counteracting Age-Related Netrin-1 Signaling Insufficiency Ameliorates Endothelial Cell Senescence and Angiogenesis Impairment.

Senescent cells that accumulate are regarded as promising therapeutic targets. However, senolytic therapy failed to achieve satisfactory results. We previously discovered that young human plasma improved vascular endothelial cell senescence, and UNC5B might be a novel intervention target. Netrin-1, as a natural ligand of UNC5B, plays roles in multiple age-related vascular disorders, but its involvement in aging is still unclear. Here, we observed a significant decrease in plasma Netrin-1 levels in old healthy subjects compared to the young. In vivo, adeno-associated-virus-mediated delivery of Netrin-1 into aged mice significantly improved functional recovery in a model of hindlimb ischemia, promoted angiogenesis in ischemic tissues, and activated the endothelial nitric oxide synthase. Furthermore, we revealed that low-dose Netrin-1 recombinant protein significantly reduced senescence-associated-ß-galactosidase-positive cells, inhibited the P53 pathway, promoted cell migration, increased tubule formation, and elevated nitric oxide production in senescent endothelial cells. However, UNC5B inhibition blocked the pro-angiogenesis effect of low-dose Netrin-1 on senescent cells or aortic rings. In summary, this study depicts that modulating Netrin-1 signaling can result in improved vascular health and Netrin-1 may have therapeutic potential for age-related ischemic diseases.

ScoreMix: A Scalable Augmentation Strategy for Training GANs With Limited Data.

Generative Adversarial Networks (GANs) typically suffer from overfitting when limited training data is available. To facilitate GAN training, current methods propose to use data-specific augmentation techniques. Despite the effectiveness, it is difficult for these methods to scale to practical applications. In this article, we present ScoreMix, a novel and scalable data augmentation approach for various image synthesis tasks. We first produce augmented samples using the convex combinations of the real samples. Then, we optimize the augmented samples by minimizing the norms of the data scores, i.e., the gradients of the log-density functions. This procedure enforces the augmented samples close to the data manifold. To estimate the scores, we train a deep estimation network with multi-scale score matching. For different image synthesis tasks, we train the score estimation network using different data. We do not require the tuning of the hyperparameters or modifications to the network architecture. The ScoreMix method effectively increases the diversity of data and reduces the overfitting problem. Moreover, it can be easily incorporated into existing GAN models with minor modifications. Experimental results on numerous tasks demonstrate that GAN models equipped with the ScoreMix method achieve significant improvements.

Memory Uncertainty Learning for Real-World Single Image Deraining.

Single image deraining has witnessed dramatic improvements by training deep neural networks on large-scale synthetic data. However, due to the discrepancy between authentic and synthetic rain images, it is challenging to directly extend existing methods to real-world scenes. To address this issue, we propose a memory-uncertainty guided semi-supervised method to learn rain properties simultaneously from synthetic and real data. The key aspect is developing a stochastic memory network that is equipped with memory modules to record prototypical rain patterns. The memory modules are updated in a self-supervised way, allowing the network to comprehensively capture rainy styles without the need for clean labels. The memory items are read stochastically according to their similarities with rain representations, leading to diverse predictions and efficient uncertainty estimation. Furthermore, we present an uncertainty-aware self-training mechanism to transfer knowledge from supervised deraining to unsupervised cases. An additional target network is adopted to produce pseudo-labels for unlabeled data, of which the incorrect ones are rectified by uncertainty estimates. Finally, we construct a new large-scale image deraining dataset of 10.2 k real rain images, significantly improving the diversity of real rain scenes. Experiments show that our method achieves more appealing results for real-world rain removal than recent state-of-the-art methods.

FGF21 at physiological concentrations regulates vascular endothelial cell function through multiple pathways.

Cardiovascular diseases are closely associated with dysfunction of vascular endothelial cells (VECs), which can be influenced by various intrinsic and extrinsic factors, including fibroblast growth factor 21 (FGF21), but the effects of serum FGF21 on VECs remain unclear. We performed a cross-sectional study nested within a prospective cohort to assess the range of physiological concentrations of fasting serum FGF21 in 212 healthy individuals. We also treated human umbilical VECs (HUVECs) with recombinant FGF21 at different concentrations. The effects of FGF21 treatment on glycolysis, nitric oxide release and reduction of intracellular reactive oxygen species were assessed. The cells were also collected for RNA transcriptomic sequencing to investigate the potential mechanisms induced by FGF21 treatment. In addition, the roles of SIRT1 in the regulation of FGF21 were evaluated by SIRT1 knockdown. The results showed that the serum FGF21 concentration in healthy individuals ranged from 15.70 to 499.96 pg/mL and was positively correlated with age and pulse wave velocity. FGF21 at 400 pg/mL was sufficient to enhance glycolysis, increase nitric oxide release and protect cells from H2O2-induced oxidative damage. The upregulated genes after FGF21 treatment were mostly enriched in metabolic pathways, whereas the downregulated genes were mostly enriched in inflammation and apoptosis signaling pathways. Moreover, SIRT1 may be involved in the regulation of some genes by FGF21. In conclusion, our data indicate that FGF21 at a level within the physiological concentration range has a beneficial effect on HUVECs and that this effect may partly depend on the regulation of SIRT1.

Association Between Plasma Fibulin-1 and Brachial-Ankle Pulse Wave Velocity in Arterial Stiffness.

Arterial stiffness forms the basis of cardiovascular diseases (CVD) and is also an independent predictor of CVD risk. Early detection and intervention of arterial stiffness are important for improving the global burden of CVD. Pulse wave velocity (PWV) is the gold standard for assessing arterial stiffness and the molecular mechanism of arterial stiffness remains to be studied. Extracellular matrix (ECM) remodeling is one of the major mechanisms of arterial stiffness. Partial quantitative changes of ECM proteins can be detected in plasma. Therefore, we examined the hypothesis that a discovery proteomic comparison of plasma proteins between high arterial stiffness (baPWV ≥ 1,400 cm/s) and normal arterial stiffness (baPWV < 1,400 cm/s) populations might identify relevant changed ECM proteins for arterial stiffness. Plasma samples were randomly selected from normal arterial stiffness (n = 6) and high arterial stiffness (n = 6) people. Isobaric tags for relative and absolute quantitation (iTRAQ) based quantitative proteomics technique was performed to find a total of 169 differentially expressed proteins (DEPs). Nine ECM proteins were included in all DEPs and were all up-regulated proteins. Fibulin-1 had the highest statistically fold-change (FC = 3.7, p < 0.0001) in the high arterial stiffness population compared with the control group during the nine ECM proteins. The expression of plasma fibulin-1 in normal arterial stiffness (n = 112) and high arterial stiffness (n = 72) populations was confirmed through enzyme-linked immunosorbent assay (ELISA). Similarly, ELISA results showed that plasma concentrations of fibulin-1 in the high arterial stiffness group were higher than those in the normal arterial stiffness group (12.69 ± 0.89 vs. 9.84 ± 0.71 µg/ml, p < 0.05). Univariate analysis of fibulin-1 with brachial-ankle pulse wave velocity (baPWV) indicated that fibulin-1 was positively correlated with baPWV in all participants (r = 0.32, p < 0.01) and a stronger positive correlation between baPWV and fibulin-1 in high arterial stiffness group (r = 0.64, p < 0.0001) was found. Multiple regression analysis of factors affecting baPWV showed that fibulin-1 was also a significant determinant of the increased ba-PWV (R 2 = 0.635, p = 0.001). Partial correlation analysis showed that baPWV increased with the growth of plasma fibulin-1(r = 0.267, p < 0.001). In conclusion, our results demonstrated that fibulin-1 is positively correlated with ba-PWV and an independent risk factor for arterial stiffness.

Carnosine attenuates vascular smooth muscle cells calcification through mTOR signaling pathway.

OBJECTIVE: Vascular calcification is prevalent in the aging population, as we know that arterial calcification is associated with aging. Recent studies have demonstrated that carnosine, a naturally occurring dipeptide, performs the treatment of aging-related diseases, such as atherosclerosis and type 2 diabetes. Here, we investigated the role of carnosine in a calcification model of vascular smooth muscle cells (VSMCs). METHODS: In this research, we used an in vitro model of VSMC calcification to investigate the role of carnosine in the progression of rat VSMC calcification. RESULTS: Carnosine treatment attenuated calcium deposition in a dose-dependent manner, detected by Alizarin Red S staining and calcium content assay. Carnosine also reduced the protein level of Runx2, bone morphogenetic protein 2 (BMP-2), and cellular reactive oxygen species (ROS) production. Further, carnosine inhibited the activation of the mammalian target of rapamycin (mTOR) pathway. CONCLUSION: Carnosine attenuated the VSMC calcification via inhibition of osteoblastic transdifferentiation and the mTOR signaling pathway.