Up-regulating microRNA-203 alleviates myocardial remodeling and cell apoptosis through down-regulating PTP1B in rats with myocardial infarction Myocardial infarction (MI) is one of cardiovascular diseases with high incidence and mortality. MicroRNAs (miRNAs), as post-transcriptional regulators of genes, are involved in many diseases, including cardiovascular diseases. The aim of present study was to determine whether miR-203 was functional in MI therapy and how it worked. Left anterior descending artery ligation and hypoxia/reoxygenation (H/R) treatment were respectively performed to obtain MI rats and hypoxia-injured H9c2 cells. Western blot and qRT-PCR were used to determine protein levels and mRNA of relevant genes, respectively. Lentivirus-mediated over-expression of miR-203 was performed to study the miR-203 functions on left ventricular remodeling, infarct size and cardiomyocyte apoptosis. Compared with the sham group, miR-203 levels were significantly decreased in MI and H/R groups. However, over-expressing miR-203 greatly improved the cardiac function, reduced infarct size in rats after MI, as well as weakened infarction-induced apoptosis by increasing Bcl-2 and reducing decreasing Bax, cleaved caspase-3 and cleaved caspase-9. In addition, PTP1B was proved as a target of miR-203 in cardiomyocytes, and it was negatively regulated by miR-203. Further experiments indicated that PTP1B over-expression could remarkably inhibit miR-203-mediated anti-apoptosis of cardiomyocytes, and alleviate protective effects of miR-203 on mitochondria after H/R treatment. Altogether, miR-203 prevented infarction-induced apoptosis by regulating PTP1B, including reducing pro-apoptosis proteins, inactivating caspase pathway and protecting mitochondria. In conclusion, miR-203 had abilities to alleviate MI-caused injury on myocardium tissues and reduce mitochondria-mediated apoptosis, which might be a potential target used for MI therapy. Correspondence author at:Zhenfei Chen, Department of Cardiovascular, the Second People Hospital of He Fei, No. 246 Heping Street, Hefei City, Anhui Province, 230021,PR. China htluyti21784408@126.com Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

The cardioprotective effects of Atorvastatin are mediated via PPARγ in Paraquat-exposed rats Background: paraquat poisoning is one of leading intoxication worldwide without an effective antidote and treatment protocol. Among the other organs, cardiotoxicity of paraquat has been frequently reported. Aim: The protective effects of atorvastatin (STN) on paraquat-induced cardiotoxicity and the role of peroxisome proliferator-activated receptors γ in the mediation of STN effects were investigated. Methods: Forty-two male Wistar rats were aliquoted into control or test groups. The animals in test groups in addition of paraquat, received saline normal (PQ), pioglitazone (PGT), atorvastatin (STN), PGT + STN, PGT + GW9662 and/or STN + GW9662 for 14 days. Results: PGT and STN lowered lipid peroxidation rate, nitric oxide concentration and activity of myeloperoxidase and CK/MB in the heart. PGT and STN protected from thiol molecules reduction and PQ-induced histopathological injuries. STN regulated the PQ-induced up-regulation of COX-II expression in the heart. All STN-related protective effects were reversed by GW9662 as PPARγ antagonist. Conclusion: These data suggest a cardioprotective effect for STN against the PQ-induced inflammation and oxidative stress. The pharmacologic approach of these findings indicate that STN via PPARγ pathway lowered the PQ-induced cardiotoxicity. Corresponding to: Hassan Malekinejad, Department of Pharmacology and Toxicology, Faculty of Pharmacy, Urmia University of Medical Sciences, Urmia, Iran Hassanmalekinejad@yahoo.com Tel: +989141496558 Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

SIRTUIN 3, ENDOTHELIAL METABOLIC REPROGRAMMING AND HEART FAILURE WITH PRESERVED EJECTION FRACTION The incidences of heart failure with preserved ejection fraction (HFpEF) are increased in the aged populations as well as diabetes and hypertension. Coronary microvascular dysfunction has been contributed to the development of HFpEF. Endothelial cells (ECs) depend on glycolysis rather than oxidative phosphorylation for generating ATP to maintain vascular homeostasis. Glycolytic metabolism has a critical role in the process of angiogenesis since endothelial cells rely on the energy produced predominantly from glycolysis for migration and proliferation. Sirtuin 3 (SIRT3) is found predominantly in mitochondria and its expression declines progressively with aging, diabetes, obesity, and hypertension. Emerging evidence indicates that endothelial SIRT33 regulates a metabolic switch between glycolysis and mitochondrial respiration. SIRT3 deficiency in EC resulted in a significant decrease in glycolysis, whereas, it exhibited higher mitochondrial respiration and more prominent production of reactive oxygen species (ROS). SIRT3 deficiency also displayed strikingly increases in acetylation of p53, EC apoptosis, and senescence. Impairment of SIRT3-mediated EC metabolism may lead to a disruption of EC/pericyte/cardiomyocyte communications and coronary microvascular rarefaction which promotes cardiomyocyte hypoxia, Titin-based cardiomyocyte stiffness, and myocardial fibrosis thus leading to a diastolic dysfunction and HFpEF. This review summarizes current knowledge of SIRT3 in EC metabolic reprograming, EC/pericyte interactions, coronary microvascular dysfunction, and HFpEF. Address for Correspondence: Jian-Xiong Chen, M.D. Department of Pharmacology and Toxicology, University of Mississippi Medical Center, 2500 North State Street, Jackson, MS, 39216 Office: 601-984-1731 Fax: 601-984-1637 Email: JChen3@umc.edu Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Sevoflurane Preconditioning Prevents Septic Myocardial Dysfunction in Lipopolysaccharide-Challenged Mice Myocardial dysfunction accompanied by severe sepsis could significantly increase the mortality rate of septic patients. This study investigated the effects and the potential mechanisms of sevoflurane preconditioning on septic myocardial dysfunction which was induced by lipopolysaccharide (LPS; from Escherichia coli O55:B5; 18 mg/kg) in mice. Results indicated that one hour after the administration, LPS induced a significant increase in cell-surface Toll-like receptor 4 (TLR4), cytoplasmic IKKα protein expression and nuclear translocation of nuclear factor kappa-B (NF-κB) protein (P<0.05), which was attenuated by preconditioning with sevoflurane. Two hours after the administration, inhalation of sevoflurane significantly reduced the serum levels of tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1β and IL-10 (P < 0.05). Twelve hours after administration, LPS caused pathological damage to the heart and elevated the serum levels of lactate dehydrogenase (LDH) and creatine kinase-MB (CK-MB; P < 0.05). Echocardiography indicated that sevoflurane preconditioning significantly improved systolic and diastolic function. The inhalation of sevoflurane inhibited increases in myeloperoxidase (MPO), macrophage inflammatory protein-2 (MIP-2), TNF-α and IL-1β levels (P < 0.05) induced by endotoxemia, whereas IL-6 release was facilitated. Sevoflurane attenuated the myocardial levels of nitric oxide (NO; P < 0.05) without an apparent influence on malondialdehyde (MDA), or superoxide dismutase (SOD; P > 0.05). In conclusion, our study indicates that exposure to 2% sevoflurane prior to LPS challenge is protective against myocardial dysfunction. Sevoflurane preconditioning may attenuate neutrophil infiltration and the release of inflammatory mediators during endotoxemia. Corresponding Author: Yalan Li, Department of Anesthesiology, First Affiliated Hospital of Jinan University, Guangzhou 510630, Guangdong, China, Phone: 0086-20-38688200 FAX: 0086-20-85222175 Email: tyalan@jnu.edu.cn, Renbin Qi, Department of Pathophysiology, Key Laboratory of State Administration of Traditional Chinese Medicine of the People's Republic of China, School of Medicine, Jinan University, Guangzhou 510632, Guangdong, China, Phone: 0086-20-85220472, FAX: 0086-20-85222175, Email: tqirb@jnu.edu.cn Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Exploration of physiological and pathophysiological implications of miRNA-143 and miRNA-145 in cerebral arteries Subarachnoid haemorrhage (SAH), is a type of haemorrhagic stroke with a high short-term mortality rate and leads to cognitive impairments that reduce the quality of life of the majority of patients. The miRNA-143/145 cluster is highly expressed in vascular smooth muscle cells (VSMC) and has been shown to be necessary for differentiation and function, as well as an important determinant for phenotypic modulation/switching of VSMCs in response to vascular injury. We aimed to determine if miRNA-143 and miRNA-145 are important regulators of phenotypical changes of VSMCs in relation to SAH, as well as establishing their physiological role in the cerebral vasculature. We applied qPCR to study ischemia-induced alterations in the expression of miRNA-143 and miRNA-145, for rat cerebral vasculature, in an ex vivo organ culture model and an in vivo subarachnoid haemorrhage model. To determine the physiological importance, we did myograph studies on basilar and femoral arteries from miRNA-143/145 KO mice. miRNA-143 and miRNA-145 are not upregulated in the vasculature following our subarachnoid haemorrhage model, despite the upregulation of miR-145 in the organ culture model. Regarding physiological function, miRNA-143 and miRNA-145 are very important for general contractility in cerebral vessels in response to depolarization, angiotensin II and endothelin-1. Applying an anti-miRNA targeting approach in SAH does not seem to be a feasible approach, as miRNA-143 and miRNA-145 are not upregulated following SAH. The KO mouse data suggest that targeting miRNA-143 and miRNA-145 would lead to a general reduced contractility of the cerebral vasculature and unwanted dedifferentiation of VSMCs. Corresponding author: Simon Topp Christensen Glostrup Research Institute Nordstjernevej 42 2600 Glostrup Denmark Tel: 0045 38633291 simon.topp.christensen.01@regionh.dk Conflict of interest: Authors report no conflict of interest. Declaration of Funding Source: This work was supported by the Lundbeck foundation, Lundbeck Grant of excellence [no. R59-A5404], the Swedish Heart Lung Foundation [no. 20130271] and the KA Wallenberg foundation [no. KAW 2016.0081]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Cardiotonic steroids induce vascular fibrosis via pressure-independent mechanism in NaCl-loaded diabetic rats Endogenous cardiotonic steroid, marinobufagenin (MBG), induces Fli1-dependent tissue fibrosis. We hypothesized that an increase in MBG initiates the development of aortic fibrosis in salt-loaded rats with type 2 diabetes mellitus (DM2) via pressure-independent mechanism. DM2 was induced by a single intraperitoneal administration of 65 mg/kg streptozotocin to neonatal (4-5 days) male Wistar rats. Eight weeks old DM2 rats received water or 1.8% NaCl (DM-NaCl) solution for 4 weeks (n=16); half of DM-NaCl rats were treated with anti-MBG monoclonal antibody (mAb) (DM-NaCl-AB) during week 4 of salt loading; control intact rats received water (n=8/group). Blood pressure (BP), MBG, erythrocyte Na/K-ATPase activity, aortic weights and levels of fibrosis markers (Fli1, PKCδ, TGFβ1, SMAD5, fibronectin, collagen-1), and sensitivity of the aortic explants to the vasorelaxant effect of sodium nitroprusside were assessed. No changes in systolic BP were observed while erythrocyte Na/K-ATPase was inhibited by 30%, plasma MBG was doubled, and aortic markers of fibrosis became elevated in DM-NaCl rats vs. control. Treatment of DM-NaCl rats with anti-MBG mAb activated Na/K-ATPase, prevented increases in aortic weights, and the levels of fibrosis markers returned to the control levels. The responsiveness of the aortic rings from DM-NaCl rats to the relaxant effect of sodium nitroprusside was reduced (EC50=29 nmol/L) vs. control rings (EC50=7 nmol/L) and was restored by anti-MBG mAb (EC50=9 nmol/L). Our results suggest that in salt-loaded diabetic rats MBG stimulates aortic collagen synthesis in a pressure-independent fashion, and that two pro-fibrotic mechanisms, Fli1-dependent and TGFβ-dependent, underlie its effects. Correspondence to Alexei Y. Bagrov, PhD, MD, Sechenov Institute of Evolutionary Physiology and Biochemistry, St. Petersburg, Russia. Fax: 7-812-552-7901; e-mail: aybagrov@gmail.com Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Drugs to inhibit the NLRP3 inflammasome: not always on-target No abstract available

GPER agonist G1 improves diastolic function and attenuates cardiac RAS activation in estrogen-deficient hypertensive rats This study was aimed to clarify differences in how specific agonists of the three estrogen receptors (ERs) influence diastolic function and the renin angiotensin system (RAS) following ovariectomy (OVX) in twenty-four spontaneously hypertensive (SHR) females undergoing bilateral OVX at 12 weeks of age. Eight weeks postsurgery, rats were randomized (n=6/group) to receive equipotent, daily treatments of one of the ER agonists (ERα agonist, PPT 94 μg/kg; ERβ agonist, DPN 58 μg/kg; GPER agonist, G1 100 μg/kg), or vehicle (peanut oil). Following 4 weeks of treatment, left ventricular (LV) function/structure and systemic/intracardiac pressure measurements were obtained by echocardiography and a fluid-filled catheter attached to a pressure transducer, respectively. Selective ER agonist treatment with G1 or PPT led to improvements in diastolic function after estrogen loss when compared to vehicle treated OVX rats. While mean arterial blood pressure was not overtly different among groups, chronic G1, but not the other ER ligands, enhanced the in vitro vasorelaxant responsiveness to acetylcholine in aortic rings. These favorable effects of G1 were further linked to reductions in cardiac ACE activity, AT1R protein expression and Ang II immunoreactivity. Activation of ERβ had no effect on cardiac function and did not alter components of the canonical cardiac RAS in comparison to vehicle-treated OVX-SHR. These data imply that of the three ERs, GPER has a unique role in preserving diastolic function and favorably modulating the cardiac RAS independent of arterial pressure. Specifically, if GPER is pharmacologically activated, it could provide a therapeutic opportunity to limit the development and/or progression of diastolic dysfunction in hypertensive women after estrogen loss. Corresponding author: Leanne Groban, MD Department of Anesthesiology Wake Forest School of Medicine Medical Center Boulevard Winston-Salem, NC 27157-1009 USA Phone: +1-336-716-4498 Email: lgroban@wakehealth.edu The authors have no conflicts of interest related to this manuscript This work was funded by grants from the National Heart Lung and Blood Institute (CMF and LG) P01-HL051952 and the National Institute on Aging (LG) AG033727 of the National Institute of Health and Conselho Nacional de Desenvolvimento Cientifico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Programa de Apoio a Núcleos de Experiência (PRONEX), Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ), Instituto Nacional de Ciência e Tecnologia (INCT-INOFAR, Proc. 465.249/2014-0). Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Role of matrix vesicles in bone-vascular cross-talk Matrix mineralization can be divided into physiological mineralization and pathological mineralization. There is a consensus among existing studies that matrix vesicles (MVs) are the starting sites of bone mineralization, and each component of MVs serves a certain function in mineralization. Additionally, ectopic MVs pathologically promote undesired calcification, the primary focus of which is the promotion of vascular calcification. However, the specific mechanisms of the actions of MVs in bone-vascular axis cross-talk have not been fully elucidated. This review summarizes the latest research in this field and explores the roles of MVs in the bone-vascular axis with the aim of generating new ideas for the prevention and treatment of vascular calcification and bone metabolic disease. Corresponding author at: Department of Cardiology, Affiliated Hospital of Jiangsu University, Zhenjiang 212001, China Tel.: +86 511 85030586. E-mail address: wangtsmc@aliyun.com (Zhongqun Wang). Jiangsu University, Zhenjiang 212001. E-mail address: 459736150@qq.com (Lele Jing). Academic degrees: Dr. Jing, Dr. Li, Dr. Sun, Dr. Bao, Miss Shao, Dr. Yan, Miss Pang, Miss Geng, Miss Zhang, Miss Wang, Dr. Wang. Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.

Association of MMP-1 (rs1799750)-1607 2G/2G and MMP-3 (rs3025058)-1612 6A/6A genotypes with coronary artery disease risk among Iranian Turks This study was conducted to evaluate the association between MMP-1 (rs1799750) -1607 1G/2G and MMP-3 (rs3025058) -1612 5A/6A polymorphisms/haplotypes and coronary artery disease (CAD) risk among Iranian Turks. Totally, 102 patients with CAD and 102 healthy subjects joined the study. Genomic DNA isolation was carried out using “salting out” method from 3-4 ml whole blood samples. The MMP-1 (-1607 2G/1G) and MMP-3 (-1612 5A/6A) promoter gene polymorphisms were detected via PCR-RFLP. Our results indicated that the frequencies of the MMP-1 (−1607) 2G alleles and 2G/2G genotypes and the MMP-3 (-1612) 6A alleles and 6A/6A genotypes were higher in CAD patients aged over 50 years than healthy controls (P < 0.05). We failed to show statistically significant differences between the CAD patients aged under 50 years and controls concerning MMP1 −1607ins/delG (1G>2G, rs1799750) and MMP-3 -1612ins/delA (5A/6A, rs3025058) polymorphisms (P > 0.05). The frequencies of MMP3/MMP1 haplotypes were not statistically different among tested groups (P > 0.05). This examination as the first study of its own kind in Iranian Turks, reported association between MMP-1 (rs1799750) -1607 2G/2G and MMP-3 (rs3025058) -1612 6A/6A genotypes and CAD risk in patients aged over 50 years. Corresponding author: Dr Morteza Bagheri, Assistant Professor, Cellular and molecular research center, Cellular and molecular medicine institute, Urmia University of Medical Sciences, Urmia, Iran. E-mail: mortezabagheri@umsu.ac.ir, Tel: +98-44-32770969, Fax: +98-44-32234125, P.O. Box: 5756115111. Copyright © 2019 Wolters Kluwer Health, Inc. All rights reserved.