RATIONALE:Diabetes mellitus is often associated with cardiovascular complications, which is the leading cause of morbidity and mortality among patients with diabetes mellitus, but little is known ...about the mechanism that connects diabetes mellitus to the development of cardiovascular dysfunction.
OBJECTIVE:We aim to elucidate the mechanism underlying hyperglycemia-induced cardiac dysfunction on a well-established db/db mouse model for diabetes mellitus and diabetic complications that lead to heart failure.
METHODS AND RESULTS:We first profiled the expression of microRNAs (miRNAs) by microarray and quantitative reverse transcription polymerase chain reaction on db/db mice and identified miR-320 as a key miRNA associated with the disease phenotype. We next established the clinical relevance of this finding by showing the upregulation of the same miRNA in the failing heart of patients with diabetes mellitus. We demonstrated the causal role of miR-320 in inducing diabetic cardiomyopathy, showing that miR-320 overexpression exacerbated while its inhibition improved the cardiac phenotype in db/db mice. Unexpectedly, we found that miR-320 acts as a small activating RNA in the nucleus at the level of transcription. By chromatin immunoprecipitation sequencing and chromatin immunoprecipitation quantitive polymerase chain reaction analysis of Ago2 (argonaute RISC catalytic component 2) and RNA polymerase II in response to miR-320 induction, we identified CD36 (fatty acid translocase) as a key target gene for this miRNA and showed that the induced expression of CD36 is responsible for increased fatty acid uptake, thereby causing lipotoxicity in the heart.
CONCLUSIONS:These findings uncover a novel mechanism for diabetes mellitus–triggered cardiac dysfunction, provide an endogenous case for small activating RNA that has been demonstrated to date only with synthetic RNAs in transfected cells, and suggest a potential strategy to develop a miRNA-based therapy to treat diabetes mellitus–associated cardiovascular complications.
Angiogenesis is involved in ischemic heart disease as well as the prognosis of heart failure (HF), and endothelial cells are the main participants in angiogenesis. In this study, we found that ...miR-221-3p is highly expressed in vascular tissue, especially in endothelial cells, and increased miR-221-3p was observed in heart tissue of HF patients and transverse aortic constriction (TAC)-induced HF mice. To explore the role of miR-221-3p in endothelial cells, microRNA (miRNA) mimics and inhibitors were employed in vitro. Overexpression of miR-221-3p inhibited endothelial cell proliferation, migration, and cord formation in vitro, while inhibition of miR-221-3p showed the opposite effect. Anti-argonaute 2 (Ago2) coimmunoprecipitation, dual-luciferase reporter assay, and western blotting were performed to verify the target of miR-221-3p. Hypoxia-inducible factor-1α (HIF-1α) was identified as a miR-221-3p target, and the adverse effects of miR-221-3p on endothelial cells were alleviated by HIF-1α re-expression. In vivo, a mouse model of hindlimb ischemia (HLI) was developed to demonstrate the effect of miR-221-3p on angiogenesis. AntagomiR-221-3p increased HIF-1α expression and promoted angiogenesis in mouse ischemic hindlimbs. Using the TAC model, we clarified that antagomiR-221-3p improved cardiac function in HF mice by promoting cardiac angiogenesis. Furthermore, serum miR-221-3p was detected to be negatively correlated with heart function in chronic heart failure (CHF) patients. Our results conclude that miR-221-3p inhibits angiogenesis of endothelial cells by targeting HIF-1α and that inhibition of miR-221-3p improves cardiac function of TAC-induced HF mice. Furthermore, miR-221-3p might be a potential prognostic marker of HF.
This study describes how miR-221-3p in endothelial cells reduces angiogenesis by inhibiting hypoxia-inducible factor-1α. Because antagonism of miR-221-3p significantly improves the cardiac function of mice with heart failure it may be a new and effective molecular target for progressing and treatment of heart failure.
•Pore structure of anammox granule was characterized with Micro-CT.•Pore distribution was observed to vary largely with increase of granule size.•Permeability was revealed to correlate negatively ...with granule size.•Permeability was found to deteriorate due to pore plugging by cell and EPS.
Anammox granular sludge bed technology has been widely applied for its attractive advantages. Efficient mass transfer is an important factor for the anammox granules to play their role. In this study, steady-state anammox granules were used to investigate the correlation between the permeability and granule size with the granule pore as pivot. The results of size distribution showed that the anammox granules could be divided into 6 groups: 200–500 µm (I), 500–1000 µm (II), 1000–1500 µm (III), 1500–2000 µm (IV), 2000–3000 µm (V) and ≥3000 µm (VI). The results of settling experiment demonstrated that the permeability of anammox granules was negatively correlated with the granule size. The fluid collection efficiency declined from 39.4% to 9.3% for granule group I to III, and further to 0 for granule group IV to VI (granule size was larger than 1.5 mm). The observation of micro-CT revealed that the pore structure of anammox granules varied significantly with the increase of granule size, forming a denser surface layer and sparser interior. The chemical analysis and microscopic observation indicated that the pore plugging of surface layer by cell proliferation and EPS secretion was the main cause for the permeability deterioration. The findings of this study will help to understand the mass transfer of anammox granules and promote the development of anammox processes.
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To improve the traditional energy production and consumption of resources, the acceleration of the development of a clean and green assembly line is highly important. Hydrogen is considered one of ...the most ideal options. The method of production of hydrogen through water splitting constitutes the most attractive research. We synthesized CoMoO4 nanofibers by electrospinning along with post-heat treatment at different temperatures. CoMoO4 nanofibers show a superior activity for hydrogen evolution reaction (HER) and only demand an overpotential of 80 mV to achieve a current density of 10 mA cm–2. In particular, the CoMoO4 catalyst also delivers excellent performances of oxygen evolution reaction (OER) in 1 M KOH, which is a more complicated process that needs extra energy to launch. The CoMoO4 nanofibers also showed a superior stability in multiple CV cycles and maintained a catalytic activity for up to 80 h through chronopotentiometry tests. This is attributed mainly to a synergistic interaction between the different metallic elements that caused the activity of CoMoO4 beyond single oxides. This approach proved that bimetallic oxides are promising for energy production.
BACKGROUND:Excessive reactive oxygen species generated in mitochondria has been implicated as a causal event in hypertensive cardiomyopathy. Multiple recent studies suggest that microRNAs (miRNAs) ...are able to translocate to mitochondria to modulate mitochondrial activities, but the medical significance of such a new miRNA function has remained unclear. Here, we characterized spontaneous hypertensive rats (SHRs) in comparison with Wistar rats, finding that micro RNA-21 (miR-21) was dramatically induced in SHRs relative to Wistar rats. We designed a series of experiments to determine whether miR-21 is involved in regulating reactive oxygen species generation in mitochondria, and if so, how induced miR-21 may either contribute to hypertensive cardiomyopathy or represent a compensatory response.
METHODS:Western blotting was used to compare the expression of key nuclear genome (nDNA)–encoded and mitochondrial genome (mtDNA)–encoded genes involved in reactive oxygen species production in SHRs and Wistar rats. Bioinformatics was used to predict miRNA targets followed by biochemical validation using quantitative real-time polymerase chain reaction and Ago2 immunoprecipitation. The direct role of miRNA in mitochondria was determined by GW182 dependence, which is required for miRNA to function in the cytoplasm, but not in mitochondria. Recombinant adeno-associated virus (type 9) was used to deliver miRNA mimic to rats via tail vein, and blood pressure was monitored with a photoelectric tail-cuff system. Cardiac structure and functions were assessed by echocardiography and catheter manometer system.
RESULTS:We observed a marked reduction of mtDNA-encoded cytochrome b (mt-Cytb) in the heart of SHRs. Downregulation of mt-Cytb by small interfering RNA in mitochondria recapitulates some key disease features, including elevated reactive oxygen species production. Computational prediction coupled with biochemical analysis revealed that miR-21 directly targeted mt-Cytb to positively modulate mt-Cytb translation in mitochondria. Circulating miR-21 levels in hypertensive patients were significantly higher than those in controls, showing a positive correlation between miR-21 expression and blood pressure. Remarkably, recombinant adeno-associated virus–mediated delivery of miR-21 was sufficient to reduce blood pressure and attenuate cardiac hypertrophy in SHRs.
CONCLUSIONS:Our findings reveal a positive function of miR-21 in mitochondrial translation, which is sufficient to reduce blood pressure and alleviate cardiac hypertrophy in SHRs. This observation indicates that induced miR-21 is part of the compensatory program and suggests a novel theoretical ground for developing miRNA-based therapeutics against hypertension.
A positive surface charge has been largely associated with nanoparticle (NP) toxicity. However, by screening a carbon NP library in macrophages, we found that a cationic charge does not ...systematically translate into toxicity. To get deeper insight into this, we carried out a comprehensive study on 5 cationic carbon NPs (NP2 to NP6) exhibiting a similar zeta (ζ) potential value (from + 20.6 to + 26.9 mV) but displaying an increasing surface charge density (electrokinetic charge, Q
from 0.23 to 4.39 µmol/g). An anionic and non-cytotoxic NP (NP1, ζ-potential = - 38.5 mV) was used as control.
The 5 cationic NPs induced high (NP6 and NP5, Q
of 2.95 and 4.39 µmol/g, respectively), little (NP3 and NP4, Q
of 0.78 and 1.35 µmol/g, respectively) or no (NP2, Q
of 0.23 µmol/g) viability loss in THP-1-derived macrophages exposed for 24 h to escalating NP dose (3 to 200 µg/mL). A similar toxicity trend was observed in airway epithelial cells (A549 and Calu-3), with less viability loss than in THP-1 cells. NP3, NP5 and NP6 were taken up by THP-1 cells at 4 h, whereas NP1, NP2 and NP4 were not. Among the 6 NPs, only NP5 and NP6 with the highest surface charge density induced significant oxidative stress, IL-8 release, mitochondrial dysfunction and loss in lysosomal integrity in THP-1 cells. As well, in mice, NP5 and NP6 only induced airway inflammation. NP5 also increased allergen-induced immune response, airway inflammation and mucus production.
Thus, this study clearly reveals that the surface charge density of a cationic carbon NP rather than the absolute value of its ζ-potential is a relevant descriptor of its in vitro and in vivo toxicity.
Cerebral ischemia is a major cause of morbidity and permanent disability. To date, no treatments for cerebral ischemia/reperfusion injury can be effectively administered beyond 4–6 h after the ...ischemic insult. Our study aimed to clarify the significance of Sirt3 during acute cerebral ischemia and explore Sirt3-targeted therapy for ischemic injuries. Upon establishing the oxygen–glucose deprivation/reperfusion (OGD/R) cell model, changes of Sirt3 protein levels and the effects of Sirt3 overexpression on primary hippocampal neurons were detected at indicated time points. Moreover, mitochondrial damage was observed in neurons upon OGD/R injury. The results showed that compared with the normoxia group, Sirt3 protein was significantly decreased in hippocampal neurons exposed to 1 h of OGD followed by 12 h of reperfusion. In addition, the reduction of Sirt3 protein levels contributed to OGD/R-induced neuronal injuries, a higher ratio of neuronal apoptosis, and extensive production of reactive oxygen species (ROS). However, all neuronal injuries were partly rescued by Sirt3 overexpression induced by lentivirus transfection. Mitochondrial morphologies were significantly impaired after OGD/R, but partly salvaged by Sirt3 overexpression. We further explored whether pharmacologically activating Sirt3 is protective for neurons, and found that treatment with honokiol (a Sirt3 agonist) after OGD exposure activated Sirt3 during reperfusion and significantly alleviated OGD/R-induced neuronal injuries. Because mitochondrial functions are essential for neuronal survival, the current results indicate that Sirt3 may be an efficient target to suppress ischemic injuries via maintenance of mitochondrial homeostasis. Our current findings shed light on a novel therapeutic strategy against subacute ischemic injuries.
Anammox granule is the key support of anammox sludge bed reactor. In this study, the anammox granules from a steady-state reactor were divided into 6 groups to investigate their dimension effects. ...The results of batch cultivation showed that the anammox granules with VMD (volume surface mean diameter) of 2.17 mm had the maximum SAA (specific anammox activity) of 399.6 ± 37.6 mg-N/(g-VSS·d). The bacterial community analysis demonstrated that Candidatus Kuenenia was the main detectable AnAOB genus in the anammox granules. Q-PCR together with flow cytometry indicated that the total number of viable AnAOB cells ascended with the increasing anammox granular size, suggesting the enhancement of nitrogen removal potential. On the contrary, the mass transfer efficiency descended with the increasing granular size, indicating the restriction of nitrogen removal performance. The maximum SAA was ascribed to the optimal match between nitrogen removal potential and mass transfer efficiency. The results of this study are helpful to comprehend the nitrogen removal capacity of anammox granules and to promote the optimization of anammox process.
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•Anammox granule with VMD of 2.17 mm demonstrated maximum SAA.•Total number of viable AnAOB rose with increase of granule size.•Mass transfer efficiency limited SAA with increase of granule size.•Maximum SAA was the match of N removal potential and mass transfer efficiency.
Selective modulation of ligand–receptor interaction is essential in targeted therapy. In this study, we design an intelligent “scan and unlock” DNA automaton (SUDA) system to equip a native ...protein‐ligand with cell‐identity recognition and receptor‐mediated signaling in a cell‐type‐specific manner. Using embedded DNA‐based chemical reaction networks (CRNs) on the cell surface, SUDA scans and evaluates molecular profiles of cell‐surface proteins via Boolean logic circuits. Therefore, it achieves cell‐specific signal modulation by quickly unlocking the protein‐ligand in proximity to the target cell‐surface to activate its cognate receptor. As a proof of concept, we non‐genetically engineered hepatic growth factor (HGF) with distinct logic SUDAs to elicit target cell‐specific HGF signaling and wound healing behaviors in multiple heterogeneous cell types. Furthermore, the versatility of the SUDA strategy was shown by engineering tumor necrotic factor‐α (TNFα) to induce programmed cell death of target cell subpopulations through cell‐specific modulation of TNFR1 signaling.
An intelligent molecular‐lock device of signaling ligand proteins, termed “scan and unlock” DNA automaton (SUDA), is presented. The SUDA employs DNA chemical reaction networks to autonomously scan and logically evaluate the cell‐surface‐protein profile of the designated cell type, selectively unlocking the signaling ligand–receptor interactions for cell‐specific signaling.
Metabolic abnormalities have been implicated as a causal event in diabetic cardiomyopathy (DCM). However, the mechanisms underlying cardiac metabolic disorder in DCM were not fully understood.
Db/db ...mice, palmitate treated H9c2 cells and primary neonatal rat cardiomyocytes were employed in the current study. Microarray data analysis revealed that PGC-1β may play an important role in DCM. Downregulation of PGC-1β relieved palmitate induced cardiac metabolism shift to fatty acids use and relevant lipotoxicity in vitro. Bioinformatics coupled with biochemical validation was used to confirm that PGC-1β was one of the direct targets of miR-30c. Remarkably, overexpression of miR-30c by rAAV system improved glucose utilization, reduced excessive reactive oxygen species production and myocardial lipid accumulation, and subsequently attenuated cardiomyocyte apoptosis and cardiac dysfunction in db/db mice. Similar effects were also observed in cultured cells. More importantly, miR-30c overexpression as well as PGC-1β knockdown reduced the transcriptional activity of PPARα, and the effects of miR-30c on PPARα was almost abated by PGC-1β knockdown.
Our data demonstrated a protective role of miR-30c in cardiac metabolism in diabetes via targeting PGC-1β, and suggested that modulation of PGC-1β by miR-30c may provide a therapeutic approach for DCM.
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