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1.
Chemistry ; : e202402635, 2024 Aug 28.
Article in English | MEDLINE | ID: mdl-39194284

ABSTRACT

For advanced synthetic intermediates or natural products with multiple unactivated and energetically similar C(sp3)-H bonds, controlling regioselectivity for the C-H activation is particularly challenging. The use of cytochrome P450 enzymes (CYPs) is a promising solution to the 'regioelectivity' challenge in remote C-H activation. Notably, CYPs and organic catalysts share a fundamental principle: they strive to control the distance and geometry between the metal reaction center and the target C-H site. Most structural analyses of the regioselectivity of CYPs are limited to the active pocket, particularly when explaining why regioselectivity could be altered by enzyme engineering through mutagenesis. However, the substructures responsible for forming the active pocket in CYPs are well known to display complex dynamic changes and substrate-induced plasticity. In this context, we highlight a comparative study of the recently reported paralogous CYPs, IkaD and CftA, which achieve different regioselectivity towards the same substrate ikarugamycin by distinct substructure conformations. We propose that substructural conformation-controlled regioselectivity might also be present in CYPs of other natural product biosynthesis pathways, which should be considered when engineering CYPs for regioselective modifications.

2.
Org Lett ; 26(8): 1677-1682, 2024 Mar 01.
Article in English | MEDLINE | ID: mdl-38363662

ABSTRACT

A known polycyclic tetramate macrolactam (aburatubolactam C, 3) and three new ones (aburatubolactams D-F, 4-6, respectively) were isolated from the marine-derived Streptomyces sp. SCSIO 40070. The absolute configuration of 3 was established by X-ray analysis. A combinatorial biosynthetic approach unveiled biosynthetic enzymes dictating the formation of distinct 5/5-type ring systems (such as C7-C14 cyclization by AtlB1 in 5 and C6-C13 cyclization by AtlB2 in 6) in aburatubolactams.


Subject(s)
Streptomyces , Cyclization
4.
J Nutr Biochem ; 124: 109528, 2024 02.
Article in English | MEDLINE | ID: mdl-37979712

ABSTRACT

Cerebral ischemia-reperfusion (I/R) injury is notably linked with folic acid (FA) deficiency. The aim of our investigation was to explore the effects and underlying mechanisms by which FA mitigates I/R, specifically through regulating the GCPII transcriptional adaptive program. Initially, we discovered that following cerebral I/R, levels of FA, methionine synthase (MTR), and methylenetetrahydrofolate reductase (MTHFR) were decreased, while GCPII expression was elevated. Secondly, administering FA could mitigate cognitive impairment and neuronal damage induced by I/R. Thirdly, the mechanism of FA supplementation involved suppressing the transcriptional factor Sp1, subsequently inhibiting GCPII transcription, reducing Glu content, obstructing cellular ferroptosis, and alleviating cerebral I/R injury. In summary, our data demonstrate that FA affords protection against cerebral I/R injury by inhibiting the GCPII transcriptional adaptive response. These findings unveil that targeting GCPII might be a viable therapeutic strategy for cerebral I/R.


Subject(s)
Brain Ischemia , Ferroptosis , Folic Acid Deficiency , Reperfusion Injury , Humans , Folic Acid/pharmacology , Folic Acid/metabolism , Hydrolases , Brain Ischemia/drug therapy , Cerebral Infarction , Reperfusion Injury/prevention & control , Reperfusion
5.
Angew Chem Int Ed Engl ; 62(51): e202310728, 2023 Dec 18.
Article in English | MEDLINE | ID: mdl-37917570

ABSTRACT

Regio- and chemoselective C-H activation at multi-positions of a single molecule is fascinating but chemically challenging. The homologous cytochrome P450 enzymes IkaD and CftA catalyze multiple C-H oxidations on the same polycyclic tetramate macrolactam (PoTeM) ikarugamycin, with distinct regio- and chemoselectivity. Herein we provide mechanistic understanding of their functional differences by solving crystal structures of IkaD and CftA in complex with ikarugamycin and unnatural substrates. Distinct conformations of the F/G region in IkaD and CftA are found to differentiate the orientation of PoTeM substrates, by causing different binding patterns with polar moieties to determine site selection, oxidation order, and chemoselectivity. Fine-tuning the polar subpocket altered the regioselectivity of IkaD, indicating that substrate re-orientation by mutating residues distal to the oxidation site could serve as an important method in future engineering of P450 enzymes.


Subject(s)
Cytochrome P-450 Enzyme System , Lactams , Cytochrome P-450 Enzyme System/metabolism , Oxidation-Reduction , Catalysis , Substrate Specificity
6.
Int J Pharm ; 645: 123389, 2023 Oct 15.
Article in English | MEDLINE | ID: mdl-37714315

ABSTRACT

Myocardial infarction (MI) is a common cardiovascular pathology that induces extensive sterile inflammation during its early stages, posing a severe threat to human health. Effectively modulating cardiac inflammation may improve post-MI outcomes. Unfortunately, owing to the side effects of therapeutic drugs and cardiac coronary artery occlusion, current MI drugs are sub-optimal for the clinical management of ischemic myocardia. Sulforaphane (SFN) has been adopted for MI treatment due to its myocardial protective effects and low toxicity. However, the targeted accumulation of SFN in infarcted areas remains challenging. Herein, porous magnetic silica nanoparticles (PMSNs) were synthesized and loaded with SFN to improve the specificity of targeted SFN delivery to infarcted areas in mouse models of MI. PMSNs loaded with SFN (PMSNs + SFN) decreased the levels of pro-inflammatory cytokines, thus leading to the improvement of cardiac function and cell survival without adverse effects. To further explore SFN's mechanisms of action in MI, a cellular (in vitro) model was established via oxygen and glucose deprivation (OGD). HSF1 and Nrf2 knockdown resulted in a decrease of SFN-induced HSP70 expression in OGD cells. Moreover, as a result of HSP70 knockdown, the pro-survival and anti-inflammatory effects of SFN were blocked in OGD cells. The level of pro-inflammatory cytokines decreased upon HSP70 overexpression, and cell survival rate increased under OGD conditions. In summary, the results confirm that PMSNs are capable of transporting SFN to infarcted areas in the myocardium, where the drug exerts cardioprotective effects against myocardial injury by up-regulating HSP70 through Nrf2/HSF1.

7.
BMC Musculoskelet Disord ; 24(1): 389, 2023 May 16.
Article in English | MEDLINE | ID: mdl-37193965

ABSTRACT

BACKGROUND: Ankylosing spondylitis (AS) is one of the most common immune-mediated arthritic diseases worldwide. Despite considerable efforts to elucidate its pathogenesis, the molecular mechanisms underlying AS are still not fully understood. METHODS: To identify candidate genes involved in AS progression, the researchers downloaded the microarray dataset GSE25101 from the Gene Expression Omnibus (GEO) database. They identified differentially expressed genes (DEGs) and functionally enriched them for analysis. They also constructed a protein-protein interaction network (PPI) using STRING and performed cytoHubba modular analysis, immune cell and immune function analysis, functional analysis and drug prediction.The results showed that DEGs were mainly associated with histone modifications, chromatin organisation, transcriptional coregulator activity, transcriptional co-activator activity, histone acetyltransferase complexes and protein acetyltransferase complexes. RESULTS: The researchers analysed the differences in expression between the CONTROL and TREAT groups in terms of immunity to determine their effect on TNF-α secretion. By obtaining hub genes, they predicted two therapeutic agents, AY 11-7082 and myricetin. CONCLUSION: The DEGs, hub genes and predicted drugs identified in this study contribute to our understanding of the molecular mechanisms underlying the onset and progression of AS. They also provide candidate targets for the diagnosis and treatment of AS.


Subject(s)
Gene Expression Profiling , Spondylitis, Ankylosing , Humans , Gene Expression Profiling/methods , Spondylitis, Ankylosing/diagnosis , Spondylitis, Ankylosing/drug therapy , Spondylitis, Ankylosing/genetics , Chromatin , Biomarkers, Tumor/genetics , Computational Biology/methods
8.
Acta Histochem ; 124(8): 151955, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36423389

ABSTRACT

BACKGROUND: Angiogenesis is an essential physiological process in the growth and metastasis of primary tumors. Ca2+ signaling is crucial for tumor angiogenesis. The purpose of this study was to detect the potential role of Ca2+ permeable transient receptor potential vanilloid-3 (TRPV3) in the angiogenesis of non-small cell lung cancer (NSCLC). METHODS: Small interfering RNA was used to down-regulate TRPV3 expression in A549 cells. A laser scanning confocal microscope was used to examine intracellular calcium concentration ([Ca2+]i). Human umbilical vein endothelial cells (HUVECs) tube formation and migration assay, Western blot, MTT and ELISA were performed to detect the potential mechanisms of TRPV3 in tumor angiogenesis. A mouse tumor xenograft model was performed to expound the effects of TRPV3 on tumor cell growth. RESULTS: Inhibition of TRPV3 reduced [Ca2+]i and protein expressions of VEGF and HIF-1α in A549 cells. Moreover, HIF-1α depletion decreased the secretion and expression of VEGF. Depletion of TRPV3 inhibited HUVECs proliferation, tube formation and migration induced by conditioned medium. And TRPV3 inhibition could decrease the volume of xenograft tumors and MVD of CD34+ cells. The expression levels of HIF-1α, VEGF and p-CaMKП in the xenograft tumors in RuR and siTRPV3 groups was reduced. CONCLUSIONS: TRPV3 calcium channel protein may play a key role in NSCLC angiogenesis. TRPV3 could promote the angiogenesis through HIF-1α-VEGF signaling pathway. Targeting TRPV3 channel protein by novel approaches would be useful for reversing NSCLC angiogenesis.


Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Humans , Mice , Animals , A549 Cells , Vascular Endothelial Growth Factor A/genetics , Signal Transduction , Human Umbilical Vein Endothelial Cells , TRPV Cation Channels/genetics
9.
Biochem Biophys Res Commun ; 605: 119-126, 2022 05 21.
Article in English | MEDLINE | ID: mdl-35316762

ABSTRACT

Myocardial ischemia/reperfusion (I/R) injury poses a significant threat to human health. High level of reactive oxygen species (ROS) and calcium overload are the foremost causes of myocardial damage in I/R. Sulforaphane (SFN) is known for its promising antioxidant effect. Whether or not SFN has myocardial protective effect against I/R is largely unknown. This study aimed to investigate if SFN can protect myocardium from I/R injury. We found that mice or cells pre-treated with SFN showed improved cardiac functions and cell survival. SFN treatment inhibited the production of inflammatory cytokines and the increase of intracellular calcium induced by hypoxia-reperfusion (H/R), while mitochondria membrane potential was effectively maintained. Transcriptome analysis showed that CaMKIIδ expression was down-regulated by SFN treatment in I/R myocardium, while CaMKIIN2, the inhibitor of CaMKII, was upregulated. Knockdown of CaMKIIN2 not only led to increased level of total CaMKIIδ and the phosphorylated CaMKIIδ but also blocked the pro-survival effect of SFN for H/R cells. Moreover, CaMKIIN2 overexpression was sufficient to suppress CaMKIIδ activation and improve cell survival under H/R. Taken together, this study demonstrated that SFN exerts cardioprotective effect toward I/R injury through upregulating CaMKIIN2 and down-regulating CaMKIIδ.


Subject(s)
Myocardial Reperfusion Injury , Animals , Apoptosis/physiology , Calcium/metabolism , Calcium-Calmodulin-Dependent Protein Kinase Type 2/metabolism , Isothiocyanates , Mice , Myocardial Reperfusion Injury/drug therapy , Myocardial Reperfusion Injury/metabolism , Myocardial Reperfusion Injury/prevention & control , Myocardium/metabolism , Myocytes, Cardiac/metabolism , Sulfoxides
10.
Biochem Biophys Res Commun ; 568: 103-109, 2021 09 03.
Article in English | MEDLINE | ID: mdl-34214874

ABSTRACT

At present, stem cell transplantation has a significant therapeutic effect on spinal cord injury (SCI), however, it is still challenging for the seed cells selection. In this study, in order to explore cells with wide neural repair potentials, we selected the pluripotent stem cells multilineage-differentiating stress-enduring (Muse) cells, inducing the in vitro differentiation of human Muse cells into neural precursor cells (Muse-NPCs) by applying neural induction medium. Here, we found induced Muse-NPCs expressed neural stem cell markers Nestin and NCAM, capable of differentiating into three types of neural cells (neuron, astrocyte and oligodendrocyte), and have certain biological functions. When Muse-NPCs were transplanted into rats suffering from T10 SCI, motor function was improved. These results provide an insight for application of Muse-NPCs in cell therapy or tissue engineering for the repair of SCI in future.


Subject(s)
Neural Stem Cells/cytology , Neural Stem Cells/transplantation , Neurogenesis , Spinal Cord Injuries/therapy , Adult , Animals , Astrocytes/cytology , Cells, Cultured , Female , Humans , Neurons/cytology , Oligodendroglia/cytology , Rats , Rats, Sprague-Dawley
11.
Brief Bioinform ; 22(4)2021 07 20.
Article in English | MEDLINE | ID: mdl-33313675

ABSTRACT

At present, computational methods for drug repositioning are mainly based on the whole structures of drugs, which limits the discovery of new functions due to the similarities between local structures of drugs. In this article, we, for the first time, integrated the features of chemical-genomics (substructure-domain) and pharmaco-genomics (domain-indication) based on the assumption that drug-target interactions are mediated by the substructures of drugs and the domains of proteins to identify the relationships between substructure-indication and establish a drug-substructure-indication network for predicting all therapeutic effects of tested drugs through only information on the substructures of drugs. In total, 83 205 drug-indication relationships with different correlation scores were obtained. We used three different verification methods to indicate the accuracy of the method and the reliability of the scoring system. We predicted all indications of olaparib using our method, including the known antitumor effect and unknown antiviral effect verified by literature, and we also discovered the inhibitory mechanism of olaparib toward DNA repair through its specific sub494 (o = C-C: C), as it participates in the low synthesis of the poly subfunction of the apoptosis pathway (hsa04210) by inhibiting the Inositol 1,4,5-trisphosphate receptor(s) (ITPRs) and hydrolyzing poly (ADP ribose) polymerases. ElectroCardioGrams of four drugs (quinidine, amiodarone, milrinone and fosinopril) demonstrated the effect of anti-arrhythmia. Unlike previous studies focusing on the overall structures of drugs, our research has great potential in the search for more therapeutic effects of drugs and in predicting all potential effects and mechanisms of a drug from the local structural similarity.


Subject(s)
Computational Biology , Databases, Factual , Drug Interactions , Drug Repositioning , Genomics , Humans , Proteins/chemistry , Proteins/metabolism
12.
Appl Soft Comput ; 98: 106897, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33199977

ABSTRACT

The sudden outbreak of novel coronavirus 2019 (COVID-19) increased the diagnostic burden of radiologists. In the time of an epidemic crisis, we hope artificial intelligence (AI) to reduce physician workload in regions with the outbreak, and improve the diagnosis accuracy for physicians before they could acquire enough experience with the new disease. In this paper, we present our experience in building and deploying an AI system that automatically analyzes CT images and provides the probability of infection to rapidly detect COVID-19 pneumonia. The proposed system which consists of classification and segmentation will save about 30%-40% of the detection time for physicians and promote the performance of COVID-19 detection. Specifically, working in an interdisciplinary team of over 30 people with medical and/or AI background, geographically distributed in Beijing and Wuhan, we are able to overcome a series of challenges (e.g. data discrepancy, testing time-effectiveness of model, data security, etc.) in this particular situation and deploy the system in four weeks. In addition, since the proposed AI system provides the priority of each CT image with probability of infection, the physicians can confirm and segregate the infected patients in time. Using 1,136 training cases (723 positives for COVID-19) from five hospitals, we are able to achieve a sensitivity of 0.974 and specificity of 0.922 on the test dataset, which included a variety of pulmonary diseases.

13.
ACS Chem Biol ; 15(3): 766-773, 2020 03 20.
Article in English | MEDLINE | ID: mdl-32118401

ABSTRACT

Totopotensamide A (TPM A, 1) is a polyketide-peptide glycoside featuring a nonproteinogenic amino acid 4-chloro-6-methyl-5,7-dihydroxyphenylglycine (ClMeDPG). The biosynthetic gene cluster (BGC) of totopotensamides (tot) was previously activated by manipulating transcription regulators in marine-derived Streptomyces pactum SCSIO 02999. Herein, we report the heterologous expression of the tot BGC in Streptomyces lividans TK64, and the production improvement of TPM A via in-frame deletion of two negative regulators totR5 and totR3. The formation of ClMeDPG was proposed to require six enzymes, including four enzymes TotC1C2C3C4 for 3,5-dihydroxyphenylglycine (DPG) biosynthesis and two modifying enzymes TotH (halogenase) and TotM (methyltransferase). Heterologous expression of the four-gene cassette totC1C2C3C4 led to production of 3,5-dihydroxyphenylglyoxylate (DPGX). The aminotransferase TotC4 was biochemically characterized to convert DPGX to S-DPG. Inactivation of totH led to a mutant accumulated a deschloro derivative TPM H1, and the ΔtotHi/ΔtotMi double mutant afforded two deschloro-desmethyl products TPMs HM1 and HM2. A hydrolysis experiment demonstrated that the DPG moiety in TPM HM2 was S-DPG, consistent with that of the TotC4 enzymatic product. These results confirmed that TotH and TotM were responsible for ClMeDPG biosynthesis. Bioinformatics analysis indicated that both TotH and TotM might act on thiolation domain-tethered substrates. This study provided evidence for deciphering enzymes leading to ClMeDPG in TPM A, and unambiguously determined its absolute configuration as S.


Subject(s)
Amino Acids/chemistry , Glycosides/biosynthesis , Glycosides/genetics , Methoxyhydroxyphenylglycol/analogs & derivatives , Methyltransferases/metabolism , Peptides, Cyclic/biosynthesis , Peptides, Cyclic/genetics , Gene Expression Regulation , Hydrolysis , Methoxyhydroxyphenylglycol/chemistry , Methylation , Molecular Structure , Multigene Family , Mutation , Recombination, Genetic , Streptomyces lividans/genetics , Sulfhydryl Compounds/chemistry , Transaminases/metabolism
14.
Org Lett ; 22(5): 1731-1735, 2020 03 06.
Article in English | MEDLINE | ID: mdl-32052979

ABSTRACT

Combinatorial biosynthesis of 5/5/6 type polycyclic tetramate macrolactams (PoTeMs) was achieved in an engineered ikarugamycin (5/6/5 type) producer by introducing a set of 5/5/6 type PoTeM biosynthetic genes from Streptomyces griseus. This study supports the idea that PoTeMs share a common polyene tetramate precursor generated by the hybrid polyketide synthase/nonribosomal peptide synthetase enzymes. The X-ray crystal structure of pactamide G (7) sets an example for resolving the absolute configuration of 5/5/6 type PoTeMs.


Subject(s)
Lactams/chemistry , Peptide Synthases/chemistry , Polyketide Synthases/genetics , Streptomyces griseus/chemistry , Lactams/metabolism , Molecular Structure , Peptide Synthases/metabolism , Polyketide Synthases/chemistry
15.
Mar Drugs ; 17(12)2019 Nov 25.
Article in English | MEDLINE | ID: mdl-31775228

ABSTRACT

Polycyclic tetramate macrolactams (PTMs) biosynthetic gene cluster are widely distributed in different bacterial types, especially in Streptomyces species. The mining of the genomic data of marine-derived Streptomyces sp. SCSIO 40010 reveals the presence of a putative PTM-encoding biosynthetic gene cluster (ptm' BGC) that features a genetic organization for potentially producing 5/5/6 type of carbocyclic ring-containing PTMs. A fermentation of Streptomyces sp. SCSIO 40010 led to the isolation and characterization of six new PTMs 1-6. Comprehensive spectroscopic analysis assigned their planar structures and relative configurations, and their absolute configurations were deduced by comparing the experimental electronic circular dichroism (ECD) spectra with the reported spectra of the known PTMs. Intriguingly, compounds 1-6 were determined to have a trans-orientation of H-10/H-11 at the first 5-membered ring, being distinct from the cis-orientation in their known PTM congeners. PTMs 1-5 displayed cytotoxicity against several cancer cell lines, with IC50 values that ranged from 2.47 to 17.68 µM.


Subject(s)
Lactams, Macrocyclic/chemistry , Streptomyces/chemistry , Streptomyces/genetics , Molecular Structure , Multigene Family , Oceans and Seas
16.
J Ovarian Res ; 11(1): 51, 2018 Jun 21.
Article in English | MEDLINE | ID: mdl-29929541

ABSTRACT

BACKGROUND: Menopause is the most important sign of aging in women, and the ovary is the organ most sensitive to aging. Quercetin is a potential antioxidant and free radical scavenger that is widely found in fruits, vegetables, and leaves. However, the effect of quercetin on ovarian aging has not been elucidated, and the mechanism underlying its antioxidative effect remains unclear. The purpose of the current study was to investigate whether quercetin protects ovarian function by decreasing oxidative stress. METHODS: In an in vivo experiment, female menopausal rats (12 months old) were intragastrically administered quercetin at three doses (12.5 mg/kg, 25 mg/kg, and 50 mg/kg) for 90 days, and the estrous cycles were determined by vaginal smearing. In an in vitro experiment, rat primary ovarian granulosa cells were cultured and treated with H2O2 (400 µM) alone or H2O2 plus quercetin at 5 µM, 20 µM, or 50 µM. The levels of the hormones estradiol (E2), progesterone (P), follicle-stimulating hormone (FSH) and luteinizing hormone (LH) were detected by radioimmunoassay. The serum levels of total antioxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-PX) and glutathione S-transferase (GST) were examined. The expression levels of the oxidative stress-related genes SOD-1, catalase (CAT) and glutathione synthetase (GSS) in the ovaries and ovarian granulosa cells were detected by Western blot. RESULTS: The in vivo results demonstrated that quercetin had no effects on ovarian morphology, hormone secretion, or the estrous cycle in menopausal rats. Although no significant changes were detected in the serum levels of T-AOC, SOD, GSH, GSH-PX, and GST between the quercetin and control groups, the mRNA and protein expression levels of the oxidative stress-related genes SOD-1, CAT and GSS in menopausal rat ovaries were increased by low-dose quercetin. Moreover, the in vitro results demonstrated that quercetin significantly rescued the decrease in cell viability by H2О2-induced oxidative stress and enhanced the H2O2-induced decrease in expression of oxidative stress-related proteins. CONCLUSIONS: Together, the results of this study indicated that quercetin increased the antioxidant capacity of the ovary by upregulating the expression of some oxidative stress-related genes both in vivo and in vitro.


Subject(s)
Antioxidants/administration & dosage , Granulosa Cells/metabolism , Ovarian Follicle/drug effects , Quercetin/administration & dosage , Animals , Catalase/metabolism , Estradiol/metabolism , Estrous Cycle/drug effects , Female , Follicle Stimulating Hormone/metabolism , Glutathione/genetics , Glutathione Peroxidase/metabolism , Granulosa Cells/drug effects , Humans , Menopause/drug effects , Ovarian Follicle/metabolism , Oxidative Stress/drug effects , Progesterone/metabolism , Rats
17.
Sci Rep ; 7: 46307, 2017 04 11.
Article in English | MEDLINE | ID: mdl-28397812

ABSTRACT

Atrial Fibrillation (AF) is common in the elderly and Sestrins (Sesns) have been suggested to prevent age-related pathologies. The aim of this study was to investigate the effects of Sesns in AF. Clinical data were collected and a small sample of atrial appendage and atrium was obtained from patients undergoing valve repairment. The expression of Sesn1, Sesn2, and Sesn3 was significantly higher in patients with permanent atrial fibrillation (PmAF) than that in sinus rhythm (SR), and further greater in the left atrium than the right in PmAF patients. Superoxide anion and malondialdehyde were enhanced and positively correlated to the protein expression of Sesn1/2/3. Reactive oxygen species (ROS) production and Ca2+ overload were significantly decreased and cell survival was enhanced by overexpression of Sesns 1/2/3 in cultured HL-1 cells. Conversely, knockdown of Sesn1/2/3 resulted in significantly increased ROS and Ca2+ overload. In addition, the overexpression of Sesn1/2 significantly reduced the proliferation of fibroblasts, as well as decreased the protein expression of collagen and fibronectin1 in angiotensin II-stimulated cardiac fibroblasts. Our study demonstrated for the first time that Sesns expression is significantly up-regulated in AF, which therefore may protect hearts against oxidative damage and atrial fibrosis.


Subject(s)
Atrial Fibrillation/metabolism , Atrial Fibrillation/pathology , Gene Expression Regulation , Heart Atria/metabolism , Heart Atria/pathology , Oxidative Stress , Angiotensin II/metabolism , Atrial Fibrillation/etiology , Atrial Fibrillation/physiopathology , Biomarkers , Calcium/metabolism , Cell Line , Cell Survival/genetics , Collagen/metabolism , Comorbidity , Female , Fibroblasts/metabolism , Fibrosis , Gene Knockdown Techniques , Heart Atria/physiopathology , Histocytochemistry , Humans , Male , Middle Aged , Reactive Oxygen Species/metabolism
18.
Cell Prolif ; 49(4): 484-93, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27302634

ABSTRACT

OBJECTIVES: Pulmonary arterial hypertension (PAH) is a fast progressing vascular disease characterized by uncontrolled cell proliferation of pulmonary artery smooth muscle cells (PASMCs). Some studies have suggested that PAH and cancers share an apoptosis-resistant state, featuring excessive cell proliferation. The miR-34 family consists of tumour-suppressive miRNAs, and its reduced expression has been reported in numerous cancers; however, its role in hypoxia-induced PAH has not been previously studied. MATERIALS AND METHODS: miR-34 family expression was evaluated in a rat model with hypoxia and in cultured hypoxic PASMCs, using real-time quantitative PCR (RT-qPCR). Function of miR-34 family was assessed by transfecting miR-34 mimics and inhibitors. Dual luciferase reporter gene assays, RT-qPCR and Western blotting were performed to validate target genes of miR-34. RESULTS: Significant down-regulation of miR-34a in hypoxic lung tissue, pulmonary artery and PASMCs was identified and then effects of miR-34a in modulating cell proliferation in human pulmonary artery smooth muscle cells (hPASMCs) was investigated in vitro. Reduction of miR-34a levels in hPASMCs caused increased proliferation and these effects were reversed by overexpression of miR-34a. miR-34a overexpression down-regulated platelet-derived growth factor receptor alpha (PDGFRA) expression, which is a key factor in PAH development. These results suggest that miR-34a is a potential regulator of proliferation in PASMCs, and that it could be used as a novel treatment strategy in PAH.


Subject(s)
Cell Proliferation , Hypoxia/metabolism , MicroRNAs/metabolism , Myocytes, Smooth Muscle/cytology , Pulmonary Artery/cytology , Receptor, Platelet-Derived Growth Factor alpha/metabolism , Animals , Apoptosis , Cell Hypoxia , Cell Line , Cyclin A/analysis , Cyclin A/metabolism , Cyclin E/analysis , Cyclin E/metabolism , DNA Damage , Down-Regulation , Humans , Hypertension, Pulmonary/genetics , Hypertension, Pulmonary/metabolism , Hypertension, Pulmonary/pathology , Hypoxia/genetics , Hypoxia/pathology , Lung/blood supply , Lung/metabolism , Lung/pathology , MicroRNAs/genetics , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/pathology , NFATC Transcription Factors/genetics , Nerve Tissue Proteins/genetics , Potassium Channels, Tandem Pore Domain/genetics , Pulmonary Artery/metabolism , Pulmonary Artery/pathology , Rats , Receptor, Platelet-Derived Growth Factor alpha/genetics , Up-Regulation
19.
Neuro Endocrinol Lett ; 37(1): 41-5, 2016.
Article in English | MEDLINE | ID: mdl-26994384

ABSTRACT

OBJECTIVE: Oxytocin (OXT) could facilitate preovulatory luteinizing hormone surge in animals and humans while brain OXT production depends on glial fibrillary acidic protein (GFAP)-associated astrocytic plasticity. Here, we examined if GFAP expressions in OXT-producing hypothalamic supraoptic nucleus (SON) correlate to special estrous stages. METHODS: 38 adult female rats were classified into diestrus, proestrus, estrus, and metestrus groups determined by vaginal smear. Rats were decapitated and the SON was dissected for detecting Fos and GFAP levels by Western Blot and immunohistochemistry. RESULTS: The result showed that Fos expression was significantly high at proestrus compared to other stages in Western blotting. No significant difference in total GFAP expression was observed between rats at different stages of the estrous cycle; however, at proestrus GFAP level at the dorsolateral portion of the SON (a region filled with OXT neurons) was significantly lower than that at the ventro-medial portion in immunohistochemistry. CONCLUSION: There is a functional correlation between supraoptic neuron activity and proestrous OXT peak during estrous cycle; it is likely that a plastic change in GFAP expression in astrocytes selectively occurs around OXT neurons at proestrus and facilitates OXT release.


Subject(s)
Astrocytes/metabolism , Estrous Cycle/metabolism , Glial Fibrillary Acidic Protein/metabolism , Supraoptic Nucleus/metabolism , Animals , Female , Immunohistochemistry , Oxytocin/metabolism , Rats , Rats, Sprague-Dawley
20.
Med Sci Monit ; 22: 678-86, 2016 Feb 29.
Article in English | MEDLINE | ID: mdl-26927633

ABSTRACT

BACKGROUND This study was designed to investigate the effects of different doses of levetiracetam on aquaporin 4 (AQP4) expression in rats after fluid percussion injury. MATERIAL AND METHODS Sprague-Dawley rats were randomly divided into 4 groups: sham operation group, traumatic brain injury group, low-dose levetiracetam group, and high-dose levetiracetam group. Brain edema models were established by fluid percussion injury, and intervened by the administration of levetiracetam. Samples from the 4 groups were collected at 2, 6, 12, and 24 h, and at 3 and 7 days after injury. Histological observation was performed using hematoxylin-eosin staining and immunohistochemical staining. AQP4 and AQP4 mRNA expression was detected using Western blot assay and RT-PCR. Brain water content was measured by the dry-wet method. RESULTS Compared with the traumatic brain injury group, brain water content, AQP4 expression, and AQP4 mRNA expression were lower in the levetiracetam groups at each time point and the differences were statistically significant (P<0.05). The intervention effects of high-dose levetiracetam were more apparent. CONCLUSIONS Levetiracetam can lessen brain edema from fluid percussion injury by down-regulating AQP4 and AQP4 mRNA expression. There is a dose-effect relationship in the preventive effect of levetiracetam within a certain extent.


Subject(s)
Aquaporin 4/genetics , Brain Edema/drug therapy , Brain Edema/genetics , Brain Injuries/complications , Brain Injuries/drug therapy , Piracetam/analogs & derivatives , Animals , Aquaporin 4/metabolism , Blotting, Western , Brain/drug effects , Brain/metabolism , Brain/pathology , Brain Edema/etiology , Brain Injuries/genetics , Dose-Response Relationship, Drug , Gene Expression Regulation/drug effects , Immunohistochemistry , Levetiracetam , Male , Percussion , Piracetam/administration & dosage , Piracetam/pharmacology , Piracetam/therapeutic use , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats, Sprague-Dawley , Real-Time Polymerase Chain Reaction , Water/metabolism
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