Inhibited histone deacetylase 3 ameliorates myocardial ischemia-reperfusion injury in a rat model by elevating microRNA-19a-3p and reducing cyclin-dependent kinase 2.

OBJECTIVE: Over the years, the roles of microRNAs (miRNAs) and histone deacetylase 3 (HDAC3) in human diseases have been investigated. This study focused on the effect of miR-19a-3p and HDAC3 in myocardial ischemia-reperfusion (I/R) injury (MIRI) by targeting cyclin-dependent kinase 2 (CDK2). METHODS: The I/R rat models were established by coronary artery ligation, which were then treated with RGFP966 (an inhibitor of HDAC3), miR-19a-3p agomir or antagomir, or silenced CDK2 to explore their roles in the cardiac function, pathological changes of myocardial tissues, myocardial infarction area, inflammatory factors and oxidative stress factors in rats with MIRI. The expression of miR-19a-3p, HDAC3, and CDK2 was determined by RT-qPCR and western blot assay, and the interaction among which was also verified by online prediction, luciferase activity assay and ChIP assay. RESULTS: The results indicated that HDAC3 and CDK2 were upregulated while miR-19a-3p was downregulated in myocardial tissues of I/R rats. The inhibited HDAC3/CDK2 or elevated miR-19a-3p could promote cardiac function, attenuate pathological changes, inflammatory reaction, oxidative stress, myocardial infarction area and apoptosis of myocardial tissues. HDAC3 mediates miR-19a-3p and CDK2 is targeted by miR-19a-3p. CONCLUSION: Inhibited HDAC3 ameliorates MIRI in a rat model by elevating miR-19a-3p and reducing CDK2, which may contribute to the treatment of MIRI.

MicroRNA-101 Protects Against Cardiac Remodeling Following Myocardial Infarction via Downregulation of Runt-Related Transcription Factor 1.

Background Myocardial infarction (MI) generally leads to heart failure and sudden death. The hearts of people with MI undergo remodeling with the features of expanded myocardial infarct size and dilated left ventricle. Many microRNAs (miRs) have been revealed to be involved in the remodeling process; however, the participation of miR-101 remains unknown. Therefore, this study aims to find out the regulatory mechanism of miR-101 in MI-induced cardiac remodeling. Methods and Results Microarray data analysis was conducted to screen differentially expressed genes in MI. The rat model of MI was established by left coronary artery ligation. In addition, the relationship between miR-101 and runt-related transcription factor 1 (RUNX1) was identified using dual luciferase reporter assay. After that, the rats injected with lentiviral vector expressing miR-101 mimic, inhibitor, or small interfering RNA against RUNX1 were used to examine the effects of miR-101 and RUNX1 on transforming growth factor ß signaling pathway, cardiac function, infarct size, myocardial fibrosis, and cardiomyocyte apoptosis. RUNX1 was highly expressed, while miR-101 was poorly expressed in MI. miR-101 was identified to target RUNX1. Following that, it was found that overexpression of miR-101 or silencing of RUNX1 improved the cardiac function and elevated left ventricular end-diastolic and end-systolic diameters. Also, miR-101 elevation or RUNX1 depletion decreased infarct size, myocardial fibrosis, and cardiomyocyte apoptosis. Moreover, miR-101 could negatively regulate RUNX1 to inactivate the transforming growth factor ß1/Smad family member 2 signaling pathway. Conclusions Taken together, miR-101 plays a protective role against cardiac remodeling following MI via inactivation of the RUNX1-dependent transforming growth factor ß1/Smad family member 2 signaling pathway, proposing miR-101 and RUNX1 as potential therapeutic targets for MI.

Triphenylphosphonium and D-α-tocopheryl polyethylene glycol 1000 succinate-modified, tanshinone IIA-loaded lipid-polymeric nanocarriers for the targeted therapy of myocardial infarction.

BACKGROUND: Cardiovascular diseases (CVDs) are the leading causes of mortality worldwide. Currently, the best treatment options for myocardial infarction focus on the restoration of blood flow as soon as possible, which include reperfusion therapy, percutaneous coronary intervention, and therapeutic thrombolytic drugs. MATERIALS AND METHODS: In the present study, we report the development of lipid-polymeric nanocarriers (LPNs) for mitochondria-targeted delivery of tanshinone IIA (TN). D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) was linked to the triphenylphosphonium (TPP) cation. The LPNs were fabricated by nanoprecipitation method. LPNs were evaluated in vitro and in vivo in comparison with free drugs and other similar nanocarriers. RESULTS: The mean diameter of TN/nanoparticles (NPs) was 89.6 nm, while that of TN/LPNs was 121.3 nm. The zeta potential of TN/NPs and TN/LPNs was -33.6 and -22.3 mV, respectively. Compared with free TN and TN/NPs, TN/LPNs exhibited significantly improved compatibility and therapeutic efficiency. In addition, the in vivo pharmacokinetics, biodistribution, and infarct therapy studies in Sprague Dawley rats showed that TPP-TPGS/TN/LPNs had better efficiency than their nonmodified TN/LPNs counterparts in all respects. CONCLUSION: These results indicated that the TPP-TPGS/TN/LPNs were promising nanocarriers for efficient delivery of cardiovascular drugs and other therapeutic agents for the treatment of CVDs.

Sevoflurane pretreatment inhibits the myocardial apoptosis caused by hypoxia reoxygenation through AMPK pathway: An experimental study.

OBJECTIVE: To study whether sevoflurane pretreatment inhibits the myocardial apoptosis caused by hypoxia reoxygenation through AMPK pathway. METHODS: H9c2 myocardial cell lines were cultured and divided into control group (C group), hypoxia reoxygenation group (H/R group), sevoflurane pretreatment + hypoxia reoxygenation group (SP group) and sevoflurane combined with Compound C pretreatment + hypoxia reoxygenation group (ComC group), and the cell proliferation activity and apoptosis rate, myocardial enzyme levels in culture medium as well as the expression of apoptosis genes and p-AMPK in cells were determined. RESULTS: p-AMPK expression in cells of H/R group was significantly lower than that of C group, SP group was significantly higher than that of H/R group; cell proliferation activity value and Bcl-2 expression in cells of H/R group were significantly lower than those of C group, SP group were significantly higher than those of H/R group, ComC group were significantly lower than those of SP group; apoptosis rate, LDH, CK and AST levels as well as the Bax and Caspase-3 expression in cells of H/R group were significantly higher than those of C group, SP group were significantly lower than those of H/R group, ComC group were significantly higher than those of SP group. CONCLUSIONS: Sevoflurane pretreatment can activate AMPK signaling pathway to inhibit the myocardial apoptosis caused by hypoxia reoxygenation.

In vitro biomimetic construction of hydroxyapatite-porcine acellular dermal matrix composite scaffold for MC3T3-E1 preosteoblast culture.

The application of porous hydroxyapatite-collagen (HAp-Collagen) as a bone tissue engineering scaffold is hindered by two main problems: its high cost and low initial strength. As a native 3-dimenssional collagen framework, purified porcine acellular dermal matrix (PADM) has been successfully used as a skin tissue engineering scaffold. Here we report its application as a matrix for the preparation of HAp to produce a bone tissue scaffold through a biomimetic chemical process. The HAp-PADM scaffold has two-level pore structure, with large channels (∼100 µm in diameter) inherited from the purified PADM microstructure and small pores (<100 nm in diameter) formed by self-assembled HAp on the channel surfaces. The obtained HAp-PADM scaffold (S15D) has a compressive elastic modulus as high as 600 kPa. The presence of HAp in sample S15D reduces the degradation rate of PADM in collagenase solution at 37°C. After 7 day culture of MC3T3-E1 pre-osteroblasts, MTT data show no statistically significant difference on pure PADM framework and HAp-PADM scaffold (p > 0.05). Because of its high strength and nontoxicity, its simple preparation method, and designable and tailorable properties, the HAp-PADM scaffold is expected to have great potential applications in medical treatment of bone defects.

[A gene chip study of "Jun Du Yan Bingzhi" on liver in a sepsis model of rat].

OBJECTIVE: To study the effect of "Jun Du Yan Bingzhi" on genic change in liver of a sepsis rat model by gene chip technique, in order to study the mechanism of the action of the drug on the gene level. METHODS: Ninety rats were randomly divided into normal control group, model group and "Jun Du Yan Bingzhi" group, with 30 rats in each group. Sepsis was reproduced by cecal ligation and puncture (CLP) method. In "Jun Du Yan Bingzhi" group the rats were treated with intraperitoneal injection of imipenem/cilastatin (0.18 g/kg), Xuebijing injection (10 ml/kg) and gavage of "Liangge San" (15 ml/kg). In the control group and model group intraperitoneal physiological saline (10 ml/kg) was given; Survival time, and 48-hour and 72-hour survival rates of every group were observed, and changes in liver genes were examined with BiostarR-40 s chip. The ratio of Cy3/Cy5 > or =2.0 or < or =0.5 was used to screen differential genes, and NCBI database was used to identity the function of differential genes. RESULTS: The 48-hour and 72-hour survival rate of "Jun Du Yan Bingzhi" group was significantly higher than that of model group (83.3% vs. 30.0%, 76.7% vs. 17.7%, both P<0.01), 305 differential genes were found in model/control groups, with up-regulation in 159, down-regulation in 146, 500 differential genes were found in "Jun Du Yan Bingzhi" group/model group, with up-regulation in 292, down-regulation in 208, model group/control group up-regulation and "Jun Du Yan Bingzhi" group/model group down-regulation were 48, model group/control group down-regulation and "Jun Du Yan Bingzhi" group/model group up-regulation were 63. CONCLUSION: "Jun Du Yan Bingzhi" can degrade the 48-hour and 72-hour death rate of sepsis rat, through control immunization related, inflammation, signal transduction transcription regulation, cell cycle, apoptosis, substance metabolism, translation/processing/modify/degradation of protein, differentiation/proliferation/growth of cell related gene, promote multisystem function of sepsis rat to recover normal.