Semaglutide inhibits ischemia/reperfusion-induced cardiomyocyte apoptosis through activating PKG/PKCε/ERK1/2 pathway.

Apoptosis is a major pathophysiological change following myocardial ischemia/reperfusion (I/R) injury. Glucagon-like peptide 1 (GLP-1) and its receptor GLP-1R are widely expressed in the cardiovascular system and GLP-1/GLP-1R activates the protein kinase G (PKG)-related signaling pathway. Therefore, this study tested whether semaglutide, a new GLP-1 analog, inhibits I/R injury-induced cardiomyocyte apoptosis by activating the PKG/PKCε/ERK1/2 pathway. We induced myocardial I/R injury in rats and hypoxia/reoxygenation (H/R) injury in H9C2 cells and detected the effects of semaglutide, a PKG analog (8-Br-cGMP), and a PKG inhibitor (KT-5823) on the PKG/PKCε/ERK1/2 pathway and cardiomyocyte apoptosis. We found that semaglutide upregulated GLP-1R levels, and both semaglutide and 8-Br-cGMP activated the PKG/PKCε/ERK1/2 pathway, inhibited myocardial infarction (MI), decreased hs-cTNT levels, increased NT-proBNP levels, and suppressed cardiomyocyte apoptosis in I/R rats and H/R H9C2 cells. However, KT-5823 exerted contrasting effects with semaglutide and 8-Br-cGMP, and KT-5823 weakened the cardioprotective effects of semaglutide. In conclusion, semaglutide inhibits I/R injury-induced cardiomyocyte apoptosis by activating the PKG/PKCε/ERK1/2 pathway. The beneficial effect of GLP-1/GLP-1R, involved in the activation of the PKG/PKCε/ERK1/2 pathway, may provide a novel treatment method for myocardial I/R injury.

[Irisin Alleviates Inflammatory Injury in Diabetic Cardiomyopathy by Regulating NF-κB Pathway].

Objective: To investigate the protective effect of irisin in diabetic cardiomyopathy (DCM) and its mechanism. Methods: A mouse model of DCM was established by high-fat diet combined with the injection of streptozotocin. The mice were assigned to a control group, a DCM group, a DCM+low-dose irisin group, a DCM+high-dose irisin group, and a DCM+pyrrolidine dithiocarbamate (PDTC) (nuclear factor [NF]-κB inhibitor) group. Then, the mice received irisin intervention for 3 weeks after successful modeling. Myocardial morphologic changes were observed by hematoxylin and eosin (HE) staining and Masson staining. The levels of serum creatine kinase (CK) and creatine kinase isoenzyme CK-MB were examined by automatic biochemical analyzer. H9c2 cells were divided into the control group, high glucose and high lipid (HG/HL) group, HG/HL+low-dose irisin group, HG/HL+high-dose irisin group, and HG/HL+PDTC group. CCK-8 assay was conducted to determine cell viability. The expression levels of tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß, and IL-6 in the myocardial tissue and the cells were determined by ELISA. In addition, nuclear translocation of NF-κB p65 protein and the protein expression level of NF-κB inhibitor protein α (IκBα) in the myocardial tissue and the cells were determined by Western blot. Results: According to the results of animal experiment, low and high doses of irisin could alleviate the pathological injury and fibrosis of myocardial tissue to varying degrees. Irisin inhibited the levels of CK, CK-MB, and inflammatory factors, up-regulated IκB protein expression, and diminished NF-κB nuclear translocation. According to the results of cell experiment, low and high doses of irisin could enhance H9c2 cell viability to varying degrees, increase the level of intracellular IκB proteins, and inhibit NF-κB p65 nuclear translocation and inflammatory factor expression. The changes in these aspects in the DCM+low-dose irisin group and the DCM+high-dose irisin group were similar to those in the DCM+PDTC group. Conclusion: Through inhibiting NF-κB p65 nuclear translocation, irisin may reduce the inflammatory response in the myocardial tissue of DCM mice and H9c2 cells of myocardial injury induced by high glucose and high fat, thereby exerting a protective effect on myocardium.

Individual Plasmonic Nanoprobes for Biosensing and Bioimaging: Recent Advances and Perspectives.

With the advent of nanofabrication techniques, plasmonic nanoparticles (PNPs) have been widely applied in various research fields ranging from photocatalysis to chemical and bio-sensing. PNPs efficiently convert chemical or physical stimuli in their local environment into optical signals. PNPs also have excellent properties, including good biocompatibility, large surfaces for the attachment of biomolecules, tunable optical properties, strong and stable scattering light, and good conductivity. Thus, single optical biosensors with plasmonic properties enable a broad range of uses of optical imaging techniques in biological sensing and imaging with high spatial and temporal resolution. This work provides a comprehensive overview on the optical properties of single PNPs, the description of five types of commonly used optical imaging techniques, including surface plasmon resonance (SPR) microscopy, surface-enhanced Raman scattering (SERS) technique, differential interference contrast (DIC) microscopy, total internal reflection scattering (TIRS) microscopy, and dark-field microscopy (DFM) technique, with an emphasis on their single plasmonic nanoprobes and mechanisms for applications in biological imaging and sensing, as well as the challenges and future trends of these fields.

Secreted frizzled-related protein 5 protects against oxidative stress-induced apoptosis in human aortic endothelial cells via downregulation of Bax.

This study was undertaken to determine the role of secreted frizzled-related protein 5 (SFRP5) in endothelial oxidative injury. Human aortic endothelial cells (HAECs) were exposed to different oxidative stimuli and examined for SFRP5 expression. The effects of SFRP5 overexpression and knockdown on cell viability, apoptosis, and reactive oxygen species production were measured. HAECs treated with angiotensin (Ang) II (1 µM) or oxidized low-density lipoprotein (oxLDL) (150 µg/mL) showed a significant increase in SFRP5 expression. Overexpression of SFRP5 significantly attenuated the viability suppression and apoptosis induction by Ang II and oxLDL, whereas the knockdown of SFRP5 exerted opposite effects. Overexpression of SFRP5 prevented ROS formation and ß-catenin activation and reduced Bax expression. Co-expression of Bax significantly reversed the anti-apoptotic effect of SFRP5 overexpression, whereas knockdown of Bax restrained Ang II- and oxLDL-induced apoptosis in HAECs. Taken together, SFRP5 confers protection against oxidative stress-induced apoptosis through inhibition of ß-catenin activation and downregulation of Bax.

Hyperuricemia Is an Independent Risk Factor for Erectile Dysfunction.

INTRODUCTION: Hyperuricemia may be related to the development of endothelial dysfunction and cardiovascular diseases. However, the association between hyperuricemia and erectile dysfunction (ED) is not currently clear. AIM: The goal of this study is to investigate the effect of hyperuricemia on erectile function and possible mechanisms. METHODS: Twenty-four 8-week-old male SD rats were randomly divided into 4 groups. Group A (control): Rats received normal saline and served as controls. Group B (hyperuricemia): rats were given oxonic acid 250 mg/kg bw/day through gastric gavage for 4 weeks. Group C (febuxostat): normal rats were treated with 5 mg/kg febuxostat through gastric gavage for 4 weeks. Group D (hyperuricemia + Febuxostat): normal rats were treated with 250 mg/kg bw/day oxonic acid and 5 mg/kg bw/day febuxostat with 1 hour interval for 4 weeks. MEASUREMENTS: The level of serum uric acid, the maximum intracavernosal pressure (ICPmax), mean arterial pressure (MAP), and the expression of endothelial nitric oxide synthase (eNOS), phospho-eNOS, neuronal NOS, Rho-associated protein kinaise (ROCK)1 and ROCK2 and the level of nitric oxide (NO) and reactive oxygen species (ROS) in cavernous tissue were determined. RESULTS: The level of serum uric acid and ROS in hyperuricemic rats was significantly higher than that in the other 3 groups (P < .05). After electrostimulation with 3 and 5 voltage, the ratio of ICPmax/MAP in hyperuricemic rats was significantly less than that in other 3 groups (P < .05), respectively. eNOS, p-eNOS, and nNOS expression in hyperuricemic rats were significantly decreased compared to the other 3 groups (P < .05), respectively. CONCLUSION: Erectile function is impaired by hyperuricemia. The decrease of eNOS, p-eNOS, and nNOS protein expression and increase of ROS in cavernous tissue may be one of the key mechanisms of ED caused by hyperuricemia.

Icariin combined with breviscapine improves the erectile function of spontaneously hypertensive rats.

INTRODUCTION: The impaired erectile response in spontaneously hypertensive rats (SHR) is caused by increased signaling of RhoA/Rho-kinase and decreased signaling of nitric oxide (NO). Icariin improves erectile function via upregulating multitargets in NO/cyclic guanosine monophosphate (NO/cGMP) pathway, which breviscapine accomplishes by downregulating RhoA/Rho-kinase pathway. AIM: To investigate the effect and mechanism of icariin combined with breviscapine on the erectile function of SHR. METHODS: Five 12-week-old male Wistar-Kyoto (WKY) rats and 20 age-matched male SHR were evenly randomized into WKY rats control group, SHR control group, icariin-treated group, breviscapine-treated group, and combined treatment group treated by vehicle, icariin, breviscapine, and icariin plus breviscapine, respectively, by gavage for four successive weeks. Maximum intracavernosal pressure/mean arterial pressure (ICPmax/MAP) and the expression of endothelial nitric oxide synthase (eNOS), neuronal nitric oxide synthase (nNOS), phosphodiesterase type 5 inhibitors (PDE5), and Rho-associated, coiled-coil containing protein kinase 1 and 2 (ROCK1 and ROCK2) in the cavernous tissues were determined. RESULTS: The ICPmax/MAP in the combined treatment group was significantly increased compared with SHR control group, icariin-treated group, and breviscapine-treated group. The expression of eNOS and nNOS was significantly higher in the combined treatment group than in SHR control group, icariin-treated group, and breviscapine-treated group (P < 0.05). The expression of PDE5 was significantly lower in the icariin-treated group than in SHR control group (P < 0.05). The expression of ROCK1 was significantly lower in the combined treatment group than in other groups (P < 0.05). The expression of ROCK2 was significantly higher in SHR control group than in WKY rats control group, icariin-treated group, and combined treatment group (P < 0.05). Among these groups, the expression of eNOS and nNOS was the strongest, and ROCK1 was the lowest in WKY rats control group. CONCLUSION: Icariin combined with breviscapine has synergistic effects on erectile function of SHR through different signal pathways.

JAK/STAT3 signaling in cardiac fibrosis: a promising therapeutic target.

Cardiac fibrosis is a serious health problem because it is a common pathological change in almost all forms of cardiovascular diseases. Cardiac fibrosis is characterized by the transdifferentiation of cardiac fibroblasts (CFs) into cardiac myofibroblasts and the excessive deposition of extracellular matrix (ECM) components produced by activated myofibroblasts, which leads to fibrotic scar formation and subsequent cardiac dysfunction. However, there are currently few effective therapeutic strategies protecting against fibrogenesis. This lack is largely because the molecular mechanisms of cardiac fibrosis remain unclear despite extensive research. The Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling cascade is an extensively present intracellular signal transduction pathway and can regulate a wide range of biological processes, including cell proliferation, migration, differentiation, apoptosis, and immune response. Various upstream mediators such as cytokines, growth factors and hormones can initiate signal transmission via this pathway and play corresponding regulatory roles. STAT3 is a crucial player of the JAK/STAT pathway and its activation is related to inflammation, malignant tumors and autoimmune illnesses. Recently, the JAK/STAT3 signaling has been in the spotlight for its role in the occurrence and development of cardiac fibrosis and its activation can promote the proliferation and activation of CFs and the production of ECM proteins, thus leading to cardiac fibrosis. In this manuscript, we discuss the structure, transactivation and regulation of the JAK/STAT3 signaling pathway and review recent progress on the role of this pathway in cardiac fibrosis. Moreover, we summarize the current challenges and opportunities of targeting the JAK/STAT3 signaling for the treatment of fibrosis. In summary, the information presented in this article is critical for comprehending the role of the JAK/STAT3 pathway in cardiac fibrosis, and will also contribute to future research aimed at the development of effective anti-fibrotic therapeutic strategies targeting the JAK/STAT3 signaling.

Zhilong Huoxue Tongyu capsule improves myocardial ischemia/reperfusion injury via the PI3K/AKT/Nrf2 axis.

INTRODUCTION: Zhilong Huoxue Tongyu Capsule (ZL) is a Chinese medicine used for the treatment of cardio-cerebral diseases. However, the pharmacological mechanisms underlying its regulation of myocardial ischemia/reperfusion injury (MI/RI) remain unclear. PURPOSE: This study aims to investigate the effects and mechanisms of ZL on MI/RI in mice. MATERIALS AND METHODS: C57BL/6J mice were randomly assigned to four groups: Sham group, I/R group, ZL group, and ZLY group. The MI/RI mouse model was established by ligation of the left anterior descending coronary artery for 30 minutes, followed by reperfusion for 120 minutes to restore blood perfusion. Cardiac function was evaluated using cardiac ultrasound. Histopathological changes and myocardial infarction area were assessed using Hematoxylin and eosin (H&E) staining and triphenyltetrazolium chloride (TTC) staining. The changes in oxidative stress- and ferroptosis-related markers were detected. RT-qPCR, Western blot, and ELISA were conducted to further explore the mechanism of ZL in improving MI/RI. RESULTS: Our findings demonstrated that ZL exerted a protective effect against MI/RI by inhibiting ferroptosis, evidenced by the upregulation of antioxidant enzymes such as GSH and GPX4, coupled with the downregulation of ACSL4, a pro-ferroptosis factor. Furthermore, ZL positively impacted the PI3K/AKT/Nrf2 pathway by promoting ATPase activities and enhancing the relative protein expression of its components. Notably, the administration of a PI3K/AKT inhibitor reversed the antioxidant and anti-ferroptosis effects of ZL to some extent, suggesting a potential role for this pathway in mediating ZL's protective effects. CONCLUSIONS: ZL protects against MI/RI-induced ferroptosis by modulating the PI3K/AKT signaling pathway, leading to increased Nrf2 expression and activation of the HO-1/GPX4 pathway. These findings shed light on the potential therapeutic mechanisms of ZL in the context of cardiovascular diseases.

Expression of aquaporin proteins in vagina of diabetes mellitus rats.

INTRODUCTION: Aquaporins (AQPs) are membrane proteins that facilitate water movement across biological membranes. Vaginal lubrication may be mediated by blood flow and other potential mechanisms related to transudation of fluid. The most common female sexual dysfunction in diabetes is inadequate vaginal lubrication. AIM: To investigate the expression of AQP1-3 in vaginal tissue of diabetes mellitus rats. METHODS: Female Sprague-Dawley rats (N = 20) were randomly divided into group A (12-week-old nondiabetic control, N = 5), group B (16-week-old nondiabetes control, N = 5), group C (12-week-old diabetes mellitus rats, N = 5), and group D (16-week-old diabetes mellitus rats, N = 5). Vaginal fluid was measured by fluid weight absorbed by cotton swabs after pelvic nerve electrostimulation and anterior vaginal tissue was dissected for determining the expression of AQP1-3 by immunohistochemical study and Western blot. MAIN OUTCOME MEASURES: The expression of AQP1-3 was determined in the vagina of diabetes mellitus rats by Western blot. RESULTS: There are no significant differences in serum estradiol concentrations of rats among these groups (P > 0.05). Vaginal fluid was significantly lower in group C (2.7 ± 0.67 mg) and group D (2.5 ± 1.03 mg) than in group A (5.74 ± 1.23 mg) and group B (5.5 ± 1.08 mg) (P < 0.05), respectively. The protein expressions of AQP1-3 were significantly lower in group C (43.40 ± 4.83, 60.60 ± 12.80, and 59.60 ± 6.95) and group D (20.81 ± 2.86, 47.80 ± 11.43, and 54.20 ± 5.26) than in group A (116.62 ± 3.21, 110.81 ± 8.044, and 108.80 ± 4.97) and group B (122.12 ± 14.54, 111.21 ± 15.07, and 106.40 ± 4.16) (P < 0.05), respectively. CONCLUSIONS: Decreased vaginal fluid in diabetes mellitus rats after electrostimulation may be partly due to estrogen-independent decreases of AQP1-3 in vaginal tissue.

Single-cell RNA sequencing of murine hearts for studying the development of the cardiac conduction system.

The development of the cardiac conduction system (CCS) is essential for correct heart function. However, critical details on the cell types populating the CCS in the mammalian heart during the development remain to be resolved. Using single-cell RNA sequencing, we generated a large dataset of transcriptomes of ~0.5 million individual cells isolated from murine hearts at six successive developmental corresponding to the early, middle and late stages of heart development. The dataset provides a powerful library for studying the development of the heart's CCS and other cardiac components. Our initial analysis identified distinct cell types between 20 to 26 cell types across different stages, of which ten are involved in forming the CCS. Our dataset allows researchers to reuse the datasets for data mining and a wide range of analyses. Collectively, our data add valuable transcriptomic resources for further study of cardiac development, such as gene expression, transcriptional regulation and functional gene activity in developing hearts, particularly the CCS.

A translational study of urine miRNAs in acute myocardial infarction.

The currently used biomarkers for acute myocardial infarction (AMI) are blood creatinine phosphokinase-muscle band (CPK-MB), troponin-T (TnT), and troponin I (TnI). However, no good biomarkers are identified in urine after AMI, because these blood protein biomarkers are difficult to be filtered into urine. In this study, the role of urine microRNAs in the diagnosis of AMI and the mechanism involved were determined. We found that urine miR-1 was quickly increased in rats after AMI with peak at 24h after AMI, in which an over 50-fold increase was demonstrated. At 7 days after AMI, the urine miR-1 level was returned to the basal level. No miR-208 was found in normal urine. In urine from rats with AMI, miR-208 was easily detected. To determine the mechanism involved, we determined the levels of heart-released miR-1 in the liver, spleen and kidney after AMI in rats and found that the kidney was an important metabolic organ. To determine the renal elimination of blood miRNAs, we isolated serum exosomes from rats after AMI and injected these exosomes into the circulating blood of normal rats. We found that the urine miR-1 was significantly increased in exosome-injected animals. Moreover, PKH67-labeled exosomes injected into circulating blood could enter into the kidney tissues and cells, as well as urine. Furthermore, the levels of urine miR-1 were significantly increased in patients with AMI. The results suggest that urine miRNAs such as miR-1 could be novel urine biomarkers for AMI.

[Urine cardiac specific microRNA-1 level in patients with ST segment elevation acute myocardial infarction].

OBJECTIVE: To observe the change of urine level of cardiac specific microRNA-1 (miR-1) in patients with ST segment elevation acute myocardial infarction (STEAMI) and investigate its potential applications. METHODS: Urine samples were collected from 20 STEAMI patients within 12 hours after STEAMI and from 20 healthy volunteers as control. Urine miR-1 concentrations were measured with real-time quantity reverse transcription-polymerase chain reaction (qRT-PCR), at the same time serum cardiac troponin I (cTnI) and MB isoenzyme of creatine kinase (CK-MB) concentrations were measured. RESULTS: Serum level of cTnI, CK-MB and urine level of miR-1 in STEAMI patients were obviously higher than those in healthy control group [cTnI in blood: 19.27±7.53 µg/L vs. 0.02±0.01 µg/L, CK-MB in blood: 93.82±12.30 µg/L vs. 0.86±0.63 µg/L, miR-1 in urine (Ct value): 45.50±4.21 vs. 52.63±4.41, P<0.05 or P<0.01]. The urine miR-1 level in patients with STEAMI had a strong correlation with serum CK-MB or cTnI when CK-MB < 300 µg/L and cTnI <50 µg/L (Ct value of urine miR-1 with blood CK-MB: r=-0.81, P<0.01; Ct value of urine miR-1 with blood cTnI: r=-0.63, P<0.05). CONCLUSIONS: The results suggest that urine miR-1 could be a novel sensitive biomarker the early diagnosis of SETAMI.

[Changes in cardiac specific microRNA-208a level in peripheral blood in ST segment elevation acute myocardial infarction patients].

OBJECTIVE: To observe serum cardiac specific microRNA-208a (miR-208a) levels in ST segment elevation acute myocardial infarction (STEAMI) patients, and to explore the role of serum miR-208a levels in the diagnosis of STEAMI. METHODS: The serum miR-208a concentrations were assessed within 12 hours after STEAMI, while 30 healthy individuals as control. Serum miR-208a concentrations were measured with real-time quantity reverse transcription-polymerase chain reaction (qRT-PCR), and serum cardiac troponin I (cTnI) or MB isoenzyme of creatine kinase (CK-MB) concentrations were measured with enzyme linked immunosorbent assay (ELISA). RESULTS: The contents of serum cTnI or CK-MB in STEAMI patients were significantly higher than those in healthy individuals (cTnI: 17.72±8.43 µg/L vs. 0.05±0.01 µg/L, CK-MB: 250.83±177.26 µg/L vs. 71.20±20.50 µg/L, both P<0.01). Serum miR-208a concentrations were detected in all individuals with STEAMI within 60 PCR cycle (0-6 hours: 44.95±4.77, 6-12 hours: 43.98±4.68), but were beyond detection for all individuals in the healthy control group. The serum miR-208a relative levels in STEAMI patients were at least more than 215 fold than that in healthy persons, compared with qRT-PCR (Ct=60) of miR-208a in healthy control persons (P<0.01). CONCLUSION: Serum miR-208a may be a new biomarker the early diagnosis of STEAMI patients.

P21­activated kinase 1 mediates angiotensin II­induced differentiation of human atrial fibroblasts via the JNK/c­Jun pathway.

Cardiac fibrosis is a common pathophysiological condition involved in numerous types of cardiovascular disease. The renin­angiotensin system, particularly angiotensin II (AngII), serves an important role in cardiac fibrosis and remodeling. Furthermore, p21­activated kinase 1 (PAK1) is a highly conserved serine/threonine protein kinase, which is abundantly expressed in all regions of the heart. However, the role of PAK1 in AngII­mediated activation of cardiac fibroblasts remains unknown. Therefore, the present study aimed to investigate the role of PAK1 in cardiac fibroblasts and its underlying mechanisms. Human cardiac fibroblasts (HCFs) were cultured and treated with PAK1 inhibitor IPA­3 or transduced with PAK1 short hairpin (sh)RNA by lentiviral particles to silence PAK1 expression levels. Subsequently, the cell proliferation and migration abilities of the HCFs were determined. Western blot analysis was used to detect the phosphorylation status of Janus kinase (JNK) and c­Jun. A Cell Counting Kit­8 assay showed that PAK1 inhibition following treatment of HCFs with 5 µM IPA­3 or PAK1­shRNA, significantly attenuated AngII­induced proliferation of fibroblasts. In addition, wound healing and Transwell migration assays demonstrated that inhibition of PAK1 significantly inhibited AngII­induced cell migration. Finally, decreased PAK1 expression levels downregulated AngII­mediated upregulation of α­smooth muscle actin (α­SMA), collagen I, phosphorylated (p)­JNK and p­c­Jun, a downstream molecule of JNK signaling. These findings indicate that PAK1 contributes to AngII­induced proliferation, migration and transdifferentiation of HCFs via the JNK/c­Jun pathway.

miR­541­3p inhibits the viability and migration of vascular smooth muscle cells via targeting STIM1.

The transformation of vascular smooth muscle cells (VSMCs) into the proliferative migratory phenotype in the plaque area contributes to stable plaque formation and facilitates the pathogenesis of atherosclerosis. Stromal interaction molecule 1 (STIM1) has been identified to promote the proliferation of VSMCs, suggesting that STIM1 may be a potent target for the prevention and treatment of atherosclerosis. Bioinformatics analysis has previously predicted STIM1 as a target of microRNA (miR)­541­3p. The present study aimed to determine the effect of the miR­541­3p/STIM1 axis on the progression of atherosclerosis in vitro. Oxidized low­density lipoprotein (ox­LDL)­treated VSMCs were used as an in vitro atherosclerosis model. Cell Counting Kit­8 and Transwell migration assays were used to analyze cell viability and migration, respectively. Reverse transcription­quantitative PCR and western blotting were applied to measure mRNA and protein expression levels, respectively. The association between miR­541­3p and STIM1 was detected using a dual luciferase gene reporter assay. The results of the present study revealed that ox­LDL treatment significantly downregulated miR­541­3p expression levels and upregulated STIM1 expression levels in VSMCs. In addition, ox­LDL stimulation enhanced cell viability and migration. The overexpression of miR­541­3p significantly reversed the ox­LDL­mediated increase in cell viability and migration, whereas the knockdown of miR­541­3p expression enhanced the ox­LDL­mediated effects. STIM1 was confirmed to be a target gene of miR­541­3p in VSMCs. The knockdown of STIM1 significantly impaired the stimulatory effects of miR­541­3p knockdown on cell viability and migration. In conclusion, the findings of the present study suggested that miR­541­3p may efficiently repress VSMC viability and migration by targeting STIM1 under the treatment of ox­LDL. These results indicated that the miR­541­3p/STIM1 axis may represent a potent target to modulate VSMC viability and migration.

Naringenin: A Promising Therapeutic Agent against Organ Fibrosis.

Fibrosis is the final common pathology of most chronic diseases as seen in the heart, liver, lung, kidney, and skin and contributes to nearly half of death in the developed countries. Fibrosis, or scarring, is mainly characterized by the transdifferentiation of fibroblasts into myofibroblasts and the excessive accumulation of extracellular matrix (ECM) secreted by myofibroblasts. Despite immense efforts made in the field of organ fibrosis over the past decades and considerable understanding of the occurrence and development of fibrosis gained, there is still lack of an effective treatment for fibrotic diseases. Therefore, identifying a new therapeutic strategy against organ fibrosis is an unmet clinical need. Naringenin, a flavonoid that occurs naturally in citrus fruits, has been found to confer a wide range of pharmacological effects including antioxidant, anti-inflammatory, and anticancer benefits and thus potentially exerting preventive and curative effects on numerous diseases. In addition, emerging evidence has revealed that naringenin can prevent the pathogenesis of fibrosis in vivo and in vitro via the regulation of various pathways that involved signaling molecules such as transforming growth factor-ß1/small mother against decapentaplegic protein 3 (TGF-ß1/Smad3), mitogen-activated protein kinase (MAPK), phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt), sirtuin1 (SIRT1), nuclear factor-kappa B (NF-κB), or reactive oxygen species (ROS). Targeting these profibrotic pathways by naringenin could potentially become a novel therapeutic approach for the management of fibrotic disorders. In this review, we present a comprehensive summary of the antifibrotic roles of naringenin in vivo and in vitro and their underlying mechanisms of action. As a food derived compound, naringenin may serve as a promising drug candidate for the treatment of fibrotic disorders.

A dataset of dual calcium and voltage optical mapping in healthy and hypertrophied murine hearts.

Pathological hypertrophy underlies sudden cardiac death due to its high incidence of occurrence of ventricular arrhythmias. The alteration of transmural electrophysiological properties in hypertrophic cardiac murine tissue has never been explored previously. In this dataset, we have for the first time conducted high-throughput simultaneous optical imaging of transmembrane potential and calcium transients (CaT) throughout the entire hypertrophic murine hearts at high temporal and spatial resolution. Using ElectroMap, we have conducted multiple parameters analysis including action potential duration/calcium transient duration, conduction velocity, alternans and diastolic interval. Voltage-calcium latency was measured as time difference between action potential and CaT peak. The dataset therefore provides the first high spatial resolution transmural electrophysiological profiling of the murine heart, allowing interrogation of mechanisms driving ventricular arrhythmias associated with pathological hypertrophy. The dataset allows for further reuse and detailed analyses of geometrical, topological and functional analyses and reconstruction of 2-dimensional and 3-dimentional models.

Curcumin affects ox-LDL-induced IL-6, TNF-α, MCP-1 secretion and cholesterol efflux in THP-1 cells by suppressing the TLR4/NF-κB/miR33a signaling pathway.

The aim of the present study was to study the molecular mechanism of how curcumin decreases the formation of ox-LDL induced human monocyte macrophage foam cells, promotes the efflux of cholesterol and reduces the secretion of inflammatory cytokines. In vitro cultured THP-1 cells were induced to become macrophages using phorbol-12-myristate-13-acetate. The cells were then pre-treated with curcumin before inducing the foam cell model by addition of oxidized low-density lipoprotein (ox-LDL). Western blot assays were used to detect expression levels of toll-like receptor (TLR)4, nuclear factor κB (NF-κB), NF-κB inhibitor α (IκBα), phosphorylated-IκBα and ATP binding cassette transporter (ABC)A1. Reverse transcription-quantitative PCR was employed to examine mRNA levels of TLR4, microRNA (miR)33a and ABCA1. ELISAs were used to detect inflammatory factors, including tumor necrosis factor (TNF)-α, monocyte chemotactic protein (MCP)-1 and interleukin (IL)-6. ox-LDL successfully induced the foam cell model, promoted phosphorylation of IκBα, promoted nuclear translocation of NF-κB, promoted the expression of TLR4 and miR33a, and promoted the secretion of TNF-α, MCP-1 and Il-6. Additionally, ox-LDL reduced the expression of ABCA1 and cholesterol efflux. However, pretreatment with curcumin increased the expression of ABCA1 and cholesterol efflux and suppressed secretion of TNF-α, MCP-1 and Il-6. TLR4 antibodies, the NF-κB blocker, PDTC, and the miR33a inhibitor also reduced the abnormal transformations induced by ox-LDL. Curcumin promoted cholesterol efflux by suppressing the TLR4/NF-κB/miR33a signaling pathway, and reduced the formation of foam cells and the secretion of inflammatory factors.

MiR-26b Suppresses the Development of Stanford Type A Aortic Dissection by Regulating HMGA2 and TGF-ß/Smad3 Signaling Pathway.

PURPOSE: Stanford type A aortic dissection (TAAD) is one of the most dangerous cardiovascular diseases. MicroRNAs (miRNAs) have been considered as potential therapeutic targets for TAAD. In this present study, we aimed to investigate the functional role and regulatory mechanism of miR-26b in TAAD development. MATERIALS AND METHODS: MiR-26b mRNA expression was detected by real-time polymerase chain reaction (RT-PCR) and protein levels were measured by Western blot. Verifying the direct target of miR-26b was used by dual luciferase assay, RT-PCR, and Western blot. Cell Counting Kit-8 (CCK-8) and TUNEL staining assays were applied for detecting rat aortic vascular smooth muscle cells (VSMCs) viability and apoptosis, respectively. RESULTS: We found that miR-26b was under-expressed in TAAD patients and closely associated with the poor prognosis of TAAD patients. Re-expression of miR-26b facilitated while knockdown of miR-26b inhibited VSMC proliferation. However, miR-26b showed the opposite effect on cell apoptosis. More importantly, high-mobility group AT-hook 2 (HMGA2) was verified as the direct target of miR-26b. Furthermore, transforming growth factor beta (TGF-ß)/Smad3 signaling pathway was involved in the development of TAAD modulated by miR-26b. CONCLUSION: miR-26b impeded TAAD development by regulating HMGA2 and TGF-ß/Smad3 signaling pathway, which provided a potential biomarker for TAAD treatment.

Interaction between eNOS gene polymorphism and current smoking on susceptibility to coronary heart disease in Chinese people.

OBJECTIVE: This study aims to explore the relation between endothelial nitric oxide synthase (eNOS) single-nucleotide polymorphisms (SNPs) and the risk of coronary heart disease (CHD). METHODS: SNPstats (online software: http://bioinfo.iconcologia.net/SNPstats) was performed to test Hardy-Weinberg equilibrium in controls. Generalized multifactor dimensionality reduction (GMDR) was adopted to screen the preferable interaction between eNOS SNPs and smoking. RESULTS: The frequency for the rs1799983-T allele was 31.1% in CHD patients, which was significantly higher than that of 19.8% in controls (P < 0.05). The frequency for the rs891512-A allele was 28.8% in cases, which was also significantly higher than that of 20.1% in controls (P < 0.05). Logistic regression analysis showed that both rs1799983-T and rs891512-A alleles were related with increased risk of CHD, and the odds ratios (ORs) [95% confidence interval (CI)] were 1.71 (1.31-2.15) and 1.57 (1.14-2.07), respectively. High-order interactions were investigated among SNPs and environmental factors using the GMDR method. The data showed that a two-locus model (rs1799983 × smoking) had a testing accuracy of 0.60 (P = 0.001). We found that current smokers with rs1799983-GT or TT within eNOS gene have the highest CHD risk, compared to never smokers with rs1799983-GG genotype, OR (95% CI) = 2.74 (1.78-3.85), after covariates adjustment for age, gender, BMI, and alcohol drinking. CONCLUSION: The rs1799983-T and rs891512-A alleles and interaction between rs1799983 and smoking were all risk factors of CHD.