Inhibition of the lncRNA DANCR attenuates cardiomyocyte injury induced by oxygen-glucose deprivation via the miR-19a-3p/MAPK1 axis.

Long noncoding RNAs (lncRNAs) have been considered as crucial regulators of acute myocardial infarction (AMI). In this study, to analyze the effect of differentiation antagonizing nonprotein coding RNA (DANCR) of lncRNA on cardiomyocyte damage in AMI, cardiomyocyte injury was induced by oxygen-glucose deprivation (OGD). Cell counting kit-8 (CCK-8) assay and flow cytometry were used to assess cell viability and apoptosis, respectively. Quantitative real-time PCR was used to measure the expression levels of DANCR and miR-19a-3p. Bioinformatics analysis and luciferase gene reporter assay were utilized to explore the relationship among DANCR, miR-19a-3p, and mitogen-activated protein kinase 1 (MAPK1). CCK-8 and TUNEL assays were used to explore the effects of DANCR alone or plus miR-19a-3p on the viability and apoptosis of OGD/R-exposed HL-1 cells. Western blot analysis was used to detect changes in the MAPK1/ERK1/2 pathway in HL-1 cells. We found that DANCR expression and miR-19a-3p level are negatively correlated as DANCR expression is increased, while miR-19a-3p level is decreased in AMI patients' serum and OGD/R-exposed HL-1 cells. DANCR knockdown increased miR-19a-3p level, and miR-19a-3p inhibition increased DANCR expression. Moreover, DANCR directly binds to miR-19a-3p. DANCR knockdown reduced viability but induced apoptosis in OGD/R-exposed HL-1 cells, while miR-19a-3p inhibition weakens these effects. Furthermore, MAPK1 is a target of miR-19a-3p. miR-19a-3p overexpression decreases MAPK1 and ERK1/2 in HL-1 cells, while miR-19a-3p inhibition increases MAPK1 and ERK1/2 in HL-1 cells. Moreover, DANCR knockdown reduces myocardium apoptosis in mice with the left anterior descending artery ligated. DANCR knockdown effectively restores myocardial cell apoptosis by regulating the miR-19a-3p/MAPK1/ERK1/2 axis.

Exosomes released from mesenchymal stem cells overexpressing microRNA-30e ameliorate heart failure in rats with myocardial infarction.

AIM: Bone marrow-derived mesenchymal stem cells (BMMSCs) exert cardioprotective effects on myocardial infarction (MI). In this investigation, we elucidated the protective effects of BMMSCs-exosomes (Exo) expressing microRNA-30e (miR-30e) against heart failure (HF) in MI rats. METHODS: First, the differentially expressed miRNAs were analyzed using a miRNA-based microarray of MI. Subsequently, we overexpressed miR-30e in rat BMMSCs to isolate exosomes. A rat model with MI was developed and treated with Exo. Next, we examined the cardiac function of the rats, followed by the myocardial tissue extraction. HE, TUNEL and Masson's staining were used to assess the protective effects of exosomes against HF in rats. Subsequently, H9C2 cells exposed to OGD were further co-cultured with Exo. We used bioinformatics to predict the target mRNA of miR-30e and verified the binding relationship. Finally, we tested the expression and role of NF-κB p65/Caspase-9 signaling in myocardial tissues and cells. RESULTS: miR-30e was poorly expressed in myocardial tissues of MI rats. Moreover, treatment of rats with Exo overexpressing miR-30e ameliorated pathological damage, cardiomyocyte apoptosis, and fibrosis in rat myocardial tissues. Furthermore, miR-30e negatively regulated LOX1 expression, which was overexpressed in the MI rats, but further Exo treatment inhibited LOX1 expression. Moreover, Exo overexpressing miR-30e impaired the NF-κB p65/Caspase-9 signaling in myocardial tissues of MI rats. The NF-κB p65/Caspase-9 signaling inhibitor repressed the apoptosis and fibrosis of cardiomyocytes as well. CONCLUSION: Exosomal miR-30e from rat BMMSCs markedly inhibited LOX1 expression, thereby downregulating the activity of the NF-κB p65/Caspase-9 signaling and ameliorating HF after MI in rats.

Efficacy and Safety of Non-recommended Dose of New Oral Anticoagulants in Patients With Atrial Fibrillation: A Systematic Review and Meta-Analysis.

Introduction: The real-world treatment of atrial fibrillation (AF) often involves the prescription of new oral anticoagulants (NOACs) using dosing both lower and higher than recommended guidelines. Our study aimed to evaluate the efficacy and safety of non-recommended dosage of NOACs in AF patients. Methods: A systematic search was performed for relevant studies across multiple electronic databases (PubMed, Embase, Cochrane Library, Clinical Trials Registry) from inception to May 1, 2021. Multicenter randomized trials and observational studies were selected with key reporting measures for inclusion involved efficacy outcomes including stroke or systemic thromboembolism along with safety endpoints assessing major or clinically relevant bleeding events. Results: A total of 11 eligible studies were included involving 48,648 patients receiving recommended dose of NOACs and 50,116 patients receiving non-recommended dosage. Compared to AF patients treated with recommended dose regimens, administration of low dose of NOACs was associated with higher risk of stroke/systemic embolism (RR = 1.24, 95% CI 1.14-1.35, P < 0.00001), but without reducing bleeding risk (RR = 1.18, 95% CI 0.91-1.53, P = 0.21) and a higher risk of all-cause mortality (RR = 1.58, 95% CI 1.25-1.99, P = 0.0001). Moreover, high dose of NOACs was associated with higher risk of stroke and systemic embolism efficacy (RR = 1.71, 95% CI 1.06-2.76, P = 0.03) and a non-significant trend to a greater risk of major or clinically relevant bleeding (RR = 1.57, 95% CI 0.96-2.58, P = 0.07). Conclusions: AF patients treated with low dose of NOACs showed equivalent safety but with worse efficacy compared with recommended dose. High dose of NOACs was not superior to recommended dose regimens in preventing stroke/systemic embolism outcomes in AF patients.

Tremella fuciformis Polysaccharides Attenuate Oxidative Stress and Inflammation in Macrophages through miR-155.

AIM: To investigate the function of Tremella fuciformis polysaccharides (TFPS) in LPS-induced inflammation and oxidative stress of macrophages. METHODS: RAW264.7 cells were pretreated with TFPS and then stimulated with 0.1 µg/ml LPS. NFκB, Akt, p38MAPK, MCP-1, and SOD-1 were analyzed by Western blotting. Cell viability was measured using MTT assays. Reactive oxygen species (ROS) production, real-time PCR, ELISA, and immunofluorescence staining were performed on RAW264.7 cells that were treated with LPS and/or TFPS to investigate the anti-inflammatory effect of TFPS. RESULTS: LPS induced inflammation and ROS production and promoted the secretion of cytokines such as TNF-α and IL-6. LPS also enhanced the nuclear translocation of NFκB, which promoted inflammation by oxidative stress. However, pretreatment with TFPS profoundly inhibited the activation of Akt, p38MAPK, and NFκB and attenuated the expression of MCP-1 in macrophages. Meanwhile, TFPS also decreased cytokine and ROS levels and attenuated cell inflammation after treatment with LPS. Moreover, miR-155, one of the key small RNAs which regulate NFκB and inflammation in macrophages, was significantly downregulated. CONCLUSION: TFPS inhibits LPS-induced oxidative stress and inflammation by inhibiting miR-155 expression and NFκB activation in macrophages, which suggests that TFPS may be a potential reagent for inhibiting the development of inflammation.

Adiponectin receptor 1 and small ubiquitin-like modifier 4 polymorphisms are associated with risk of coronary artery disease without diabetes.

BACKGROUND: The genes encoding adiponectin receptor 1 (ADIPOR1) and small ubiquitin-like modifier 4 (SUMO4) have been linked to anti-atherogenic effects, but little is known about whether polymorphisms in the two genes, acting separately or interacting, affect risk of coronary artery disease (CAD) without diabetes. METHODS: We genotyped 200 CAD patients without diabetes and 200 controls without CAD or diabetes at three single-nucleotide polymorphisms (SNPs) in ADIPOR1 and one SNP in SUMO4, which were chosen based on previous studies. Potential associations were also explored between these SNPs and clinical characteristics of CAD without diabetes. RESULTS: Risk alleles at three SNPs in ADIPOR1 (rs7539542-G, rs7514221-C and rs3737884-G) and the G allele at SNP rs237025 in SUMO4 significantly increased risk of CAD without diabetes, with ORs ranging from 1.79 to 4.44. Carriers of any of these four risk alleles showed similar adverse clinical characteristics. Compared with individuals with a CC or GC genotype, those with a GG genotype at rs3737884 were at significantly higher risk of CAD that affected the left anterior descending coronary artery (OR: 6.77, P = 0.009), the right coronary artery (OR: 4.81, P = 0.028) or a relatively large number of vessels (P = 0.04). Individuals carrying a risk allele at one or more of the three SNPs in ADIPOR1 as well as a risk allele at the SNP in SUMO4 were at significantly higher risk of CAD without diabetes than individuals not carrying any risk alleles (OR: 5.82, 95% CI: 1.23-27.7, P = 0.013). CONCLUSIONS: SNPs in ADIPOR1 and SUMO4 are associated with elevated risk of CAD without diabetes, and SNPs in the two genes may interact to jointly affect disease risk.

Impact of attenuated plaques on TIMI grade flow and clinical outcomes of coronary artery disease patients: a systematic review and meta analysis.

BACKGROUND: Plaques with a large necrotic core or lipid pool and thin-cap fibroatheroma manifest as attenuated plaques on intravascular ultrasound (IVUS). Their impact on TIMI grade flow and clinical outcomes remains undefined. We performed a systematic review and meta-analysis to summarize the association between attenuated plaque and distal embolization and clinical outcomes of coronary artery disease (CAD) from pooled data of published eligible cohort studies. METHODS: We searched the literature on TIMI grade flow and clinical outcomes on PubMed, Ovid, EMBASE, the Cochrane Library, CNKI and WanFang databases. Study heterogeneity and publication bias were estimated. RESULTS: A total of 3,833 patients were enrolled in nine studies. Five studies investigated TIMI grade flow and attenuated plaques. They revealed no difference in TIMI grade flow before percutaneous coronary intervention (PCI) between the attenuated and non-attenuated plaque group (RR =1.25; 95% CI: 0.65 to 2.41; P=0.50). After balloon dilation and stent implantation, the incidence of TIMI 0~2 grade flow in the attenuated plaque group was statistically significant higher than that of the non-attenuated plaque group (RR =4.73; 95% CI: 3.03 to 7.40; P<0.001). Five other studies investigated major cardiovascular events (MACEs) and attenuated plaques and found no difference in MACE rates within three years of follow up. CONCLUSIONS: Our study presents the evidence that plaque with ultrasound signal attenuation would induce slow/no reflow phenomenon and distal embolization during PCI, but this appearance has no impact on MACE rates within three years.

Analysis of the Relationship Between ADIPOR1 Variants and the Susceptibility of Chronic Metabolic Diseases in a Northeast Han Chinese Population.

OBJECTIVE: Shared genetic variants in ADIPOR1 have been identified as closely related to coronary artery disease (CAD), type 2 diabetes (T2D), and T2D with CAD susceptibility, suggesting that these variants are strong candidates for the common soil hypothesis. Therefore, it is essential to analyze the relationship between ADIPOR1 variants and the susceptibility to CAD, T2D, and T2D with CAD in other populations. MATERIALS AND METHODS: A case-control study was conducted which included three case cohorts [CAD (n = 316), T2D (n = 295), T2D with CAD (n = 302)], and a control cohort (n = 268) from a population in northeast China. Six ADIPOR1 single-nucleotide polymorphisms were genotyped by high-resolution melting and polymerase chain reaction-restriction fragment length polymorphism. RESULTS: We confirmed that the shared variant, rs3737884*G, in ADIPOR1 is associated with CAD, T2D, and T2D with CAD (p-value range: 6.54E-6-1.82E-5, odds ratio [OR] range: 1.770-1.844) and that rs16850797*C is associated with T2D and T2D with CAD (p-value range: 0.001-0.001, OR range: 1.529-1.571). We also found that a novel shared variant, rs7514221*C, is associated with an increased susceptibility to CAD, T2D, and T2D with CAD (p-value range: 0.002-0.004, OR range: 1.194-2.382) in this population. CONCLUSIONS: ADPOR1 variants, rs3737884*G and rs7514221*C, may be shared risk factors associated with CAD, T2D, and T2D with CAD in a population of northeast China.

SIRT1 suppresses doxorubicin-induced cardiotoxicity by regulating the oxidative stress and p38MAPK pathways.

BACKGROUND: SIRT1, which belongs to the Sirtuin family of NAD-dependent enzymes, plays diverse roles in aging, metabolism, and disease biology. It could regulate cell survival and has been shown to be a protective factor in heart function. Hence, we verified the mechanism by which SIRT1 regulates doxorubicin induced cardiomyocyte injury in vivo and in vitro. METHODS: We analyzed SIRT1 expression in doxorubicin-induced neonatal rat cardiomyocyte injury model and adult mouse heart failure model. SIRT1 was over-expressed in cultured neonatal rat cardiomyocyte by adenovirus mediated gene transfer. SIRT1 agonist resveratrol was used to treat the doxorubicin-induced heart failure mouse model. Echocardiography, reactive oxygen species (ROS) production, TUNEL, qRT-PCR, and Western blotting were performed to analyze cell survival, oxidative stress, and inflammatory signal pathways in cardiomyocytes. RESULTS: SIRT1 expression was down-regulated in doxorubicin induced cardiomocyte injury, accompanied by elevated oxidative stress and cell apoptosis. SIRT1 over-expression reduced doxorubicin induced cardiomyocyte apoptosis with the attenuated ROS production. SIRT1 also reduced cell apoptosis by inhibition of p38MAPK phosphorylation and caspase-3 activation. The SIRT1 agonist resveratrol was able to prevent doxorubicin-induced heart function loss. Moreover, the SIRT1 inhibitor niacinamide could reverse SIRT1's protective effect in cultured neonatal rat cardiomyocytes. CONCLUSIONS: These results support the role of SIRT1 as an important regulator of cardiomyocyte apoptosis during doxorubicin-induced heart injury, which may represent a potential therapeutic target for doxorubicin-induced cardiomyopathy.

Identification of susceptibility variants in ADIPOR1 gene associated with type 2 diabetes, coronary artery disease and the comorbidity of type 2 diabetes and coronary artery disease.

OBJECTIVE: Adiponectin receptor 1 (encoded by ADIPOR1) is one of the major adiponectin receptors, and plays an important role in glucose and lipid metabolism. However, few studies have reported simultaneous associations between ADIPOR1 variants and type 2 diabetes (T2D), coronary artery disease (CAD) and T2D with CAD. Based on the "common soil" hypothesis, we investigated whether ADIPOR1 polymorphisms contributed to the etiology of T2D, CAD, or T2D with CAD in a Northern Han Chinese population. METHODS: Our multi-disease comparison study enrolled 657 subjects, including 165 with T2D, 173 with CAD, 174 with both T2D and CAD (T2D+CAD), and 145 local healthy controls. Six ADIPOR1 single nucleotide polymorphisms (SNPs) were genotyped and their association with disease risk was analyzed. RESULTS: Multi-case-control comparison identified two ADIPOR1 variants: rs3737884-G, which was simultaneously associated with an increased risk of T2D, CAD, and T2D+CAD (P-value range, 9.80×10(-5)-6.30×10(-4); odds ratio (OR) range: 1.96-2.42) and 16850797-C, which was separately associated with T2D and T2D+CAD (P-value range: 0.007-0.014; OR range: 1.71-1.77). The risk genotypes of both rs3737884 and 16850797 were consistently associated with common metabolic phenotypes in all three diseases (P-value range: 4.81×10(-42)-0.001). We observed an increase in the genetic dose-dependent cumulative risk with increasing risk allele numbers in T2D, CAD and T2D+CAD (P trend from 1.35×10(-5)-0.002). CONCLUSIONS: Our results suggest that ADIPOR1 risk polymorphisms are a strong candidate for the "common soil" hypothesis and could partially contribute to disease susceptibility to T2D, CAD, and T2D with CAD in the Northern Han Chinese population.

[Association between SUMO4 polymorphisms and coronary artery disease with and without type 2 diabetes mellitus].

OBJECTIVE: To assess the role of small ubiquitin-like modifier 4 (SUMO4) gene polymorphisms (rs237025, rs237024 and rs600739) in the susceptibility to coronary artery disease (CAD) with and without type 2 diabetes mellitus (T2DM) in Chinese Han ethnic population in Beijing. METHODS: In this case-control study, 558 subjects with angiography-proven CAD were divided into two groups according to the WHO 1999 criteria: 369 with normal glucose tolerance (CAD group) and 189 with T2DM (T2DM+ CAD group). Meanwhile 500 healthy subjects free of T2DM and CAD were selected as normal controls (control group). Allelic and genotypic distributions of the three single nucleotide polymorphisms (SNPs) were determined with polymerase chain reaction-high resolution melting curve (PCR-HRM) and gene sequencing. Clinical and biochemical data were compared among carriers of different genotypes through a stratified analysis. RESULTS: No significant difference was found in the distribution of genotypes and alleles of each SNP between different groups (P> 0.05). Nevertheless, stratified analysis indicated a significant difference in plasma triglycerides (rs237025) and body mass index (rs600739) among individuals of different genotypes from the T2DM+ CAD group (P= 0.020 and P= 0.049, respectively). Multiple comparison also indicated that GG genotype of rs237025 had a higher level of plasma triglycerides than AA genotype (P< 0.01). CONCLUSION: No association between SUMO4 gene polymorphisms and CAD with and without T2DM was detected. Such polymorphisms may not be a risk factor for Chinese Han ethnic patients in Beijing.

[Association between SUMO4 polymorphisms and type 2 diabetes mellitus].

This study investigated the association between small ubiquitin-like modifier 4 (SUMO4) gene polymorphisms and type 2 diabetes mellitus (T2DM) in Chinese Han of Beijing area. Using the case-control method, we included 404 T2DM patients in T2DM group and 500 age- and gender- matched healthy subjects in control group. We detected the distribution of alleles and genotypes of the three single nucleotide polymorphisms (SNPs, rs237025, rs237024 and rs600739) with the polymerase chain reaction-high resolution melting curve (PCR-HRM) combined with gene sequencing, analysed the differences of glycosylated hemoglobin A1c (HbA1c) among different genotypes carriers in T2DM group, and conducted a haplotype analysis. In this study, the results showed that the frequency of the G allele of rs237025 was significantly higher in T2DM group than that of control group (0.334 vs. 0.282, P = 0.017). Compared with control group, the GA genotype carriers of T2DM patients had 1.563 times more susceptibility to T2DM [P =0.001; odds ratio (OR), 1.563; 95% confidence interval (CI), 1.189-2.053]. Meanwhile, the G allele carriers (GG+GA) of T2DM patients had 1.525 times more susceptibility to T2DM in the dominant model (GG+GA vs. AA, P = 0.002; OR,1.525; 95% CI,1.169-1.989). However, as for rs237024 and rs600739, no significant differences were found in the distribution of the genotypes and alleles between two groups (P >0.05).Although our study didn't observe any statistically significant results, we found that T2DM patients with GG and GA genotypes of rs237025, TT genotype of rs237024 and GG genotype of rs600739 had a higher level of HbA1c than counterparts in control group. In addition, the AAC, AGC and GGT haplotypes might contribute to susceptibility to T2DM (OR>1) , while the AAT and GAC haplotypes might be considered as protective factors against T2DM (OR<1). The results suggested that rs237025 polymorphisms was associated with susceptibility to T2DM, but rs237024 and rs600739 were not.