Traumatic injury of the central nervous system (CNS) including brain and spinal cord remains a leading cause of morbidity and disability in the world. Delineating the mechanisms underlying the secondary and persistent injury versus the primary and transient injury has been drawing extensive attention for study during the past few decades. The sterile neuroinflammation during the secondary phase of injury has been frequently identified substrate underlying CNS injury, but as of now, no conclusive studies have determined whether this is a beneficial or detrimental role in the context of repair. Recent pioneering studies have demonstrated the key roles for the innate and adaptive immune responses in regulating sterile neuroinflammation and CNS repair. Some promising immunotherapeutic strategies have been recently developed for the treatment of CNS injury. This review updates the recent progress on elucidating the roles of the innate and adaptive immune responses in the context of CNS injury, the development and characterization of potential immunotherapeutics, as well as outstanding questions in this field.
Adaptive Immunity ; Astrocytes ; physiology ; Brain Injuries, Traumatic ; immunology ; therapy ; Histone Deacetylases ; therapeutic use ; Humans ; Immunity, Innate ; immunology ; Immunotherapy ; methods ; Inflammasomes ; drug effects ; physiology ; Macrophage Activation ; Spinal Cord Injuries ; immunology ; therapy

Background: Valproic acid (VPA) exposure during pregnancy has been proven to contribute to congenital heart disease (CHD). Our previous findings implied that disruption of planar cell polarity (PCP) signaling pathway in cardiomyocytes might be a factor for the cardiac teratogenesis of VPA. In addition, the teratogenic ability of VPA is positively correlated to its histone deacetylase (HDAC) inhibition activity. This study aimed to investigate the effect of the VPA on cardiac morphogenesis, HDAC1/2/3, and PCP key genes (Vangl2/Scrib/Rac1), subsequently screening out the specific HDACs regulating PCP pathway. Methods: VPA was administered to pregnant C57BL mice at 700 mg/kg intraperitoneally on embryonic day 10.5. Dams were sacrificed on E15.5, and death/absorption rates of embryos were evaluated. Embryonic hearts were observed by hematoxylin-eosin staining to identify cardiac abnormalities. H9C2 cells (undifferentiated rat cardiomyoblasts) were transfected with Hdac1/2/3 specific small interfering RNA (siRNA). Based on the results of siRNA transfection, cells were transfected with Hdac3 expression plasmid and subsequently mock-treated or treated with 8.0 mmol/L VPA. Hdac1/2/3 as well as Vangl2/Scrib/Rac1 mRNA and protein levels were determined by real-time quantitative polymerase chain reaction and Western blotting, respectively. Total HDAC activity was detected by colorimetric assay. Results: VPA could induce CHD (P < 0.001) and inhibit mRNA or protein expression of Hdac1/2/3 as well as Vangl2/Scrib in fetal hearts, in association with total Hdac activity repression (all P < 0.05). In vitro, Hdac3 inhibition could significantly decrease Vangl2/Scrib expression (P < 0.01), while knockdown of Hdac1/2 had no influence (P > 0.05); VPA exposure dramatically decreased the expression of Vanlg2/Scrib together with Hdac activity (P < 0.01), while overexpression of Hdac3 could rescue the VPA-induced inhibition (P > 0.05). Conclusion VPA could inhibit Hdac1/2/3, Vangl2/Scrib, or total Hdac activity both in vitro and in vivo and Hdac3 might participate in the process of VPA-induced cardiac developmental anomalies.
Animals ; Cell Polarity ; Enzyme Inhibitors ; adverse effects ; Female ; Fetal Heart ; embryology ; Heart Defects, Congenital ; chemically induced ; physiopathology ; Histone Deacetylase Inhibitors ; Histone Deacetylases ; drug effects ; physiology ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins ; Pregnancy ; Rats ; Transfection ; Valproic Acid ; adverse effects

To investigate the expression of metastasis tumor-associated protein 2 (MTA2) in cervical squamous carcinoma and its relationship with prognosis.
 Methods: Immunohistochemistry and real-time PCR were performed to determine the expression and distribution of MTA2 mRNA and protein in normal cervical tissue, cervical intraepithelial neoplasia (CIN) and cervical squamous carcinomas tissues, then its relationship with clinical pathological factors and prognosis was analyzed.
 Results: The positive rate of MTA2 protein in normal cervical tissue, CIN and cervical squamous cell carcinomas tissues were 0, 30.0%, 73.4%, respectively. The positive rate was associated with international federation of gynecology and obstetrics (FIGO) stage and lymph node metastasis, whereas there was no correlation with the age of patients or the degree of tumor differentiation. The expression of MTA2 mRNA in normal cervical tissue, CIN and cervical squamous carcinomas tissues was 0.437±0.028, 0.737±0.102 and 1.172±0.068, respectively. The positive rate was associated with FIGO stage and lymph node metastasis, whereas there was no correlation with the age of patients or the degree of tumor differentiation. The result of survival analysis showed poor overall survival time in the patients with high expression of MTA2. Multivariate COX proportional hazards model showed that the positive expression of MTA2 protein, FIGO stage and the metastasis of lymph node were independent prognostic factors for unfavorable clinical outcome of cervical cancer.
 Conclusion: The positive expression of MTA2 was closely related to the development, invasion and metastasis of cervical squamous cell carcinomas. The positive expression of MTA2 protein, FIGO stage and the metastasis of lymph node were independent prognostic factors for unfavorable clinical outcome of cervical cancer. The expression of MTA2 could be used as a potential molecular marker in evaluating the prognosis of cervical squamous cell carcinomas.
Biomarkers, Tumor ; genetics ; Carcinoma, Squamous Cell ; genetics ; mortality ; Cervical Intraepithelial Neoplasia ; genetics ; Female ; Gene Expression ; physiology ; Histone Deacetylases ; physiology ; Humans ; Immunohistochemistry ; Lymph Nodes ; Lymphatic Metastasis ; genetics ; Neoplasm Staging ; Prognosis ; Proportional Hazards Models ; Real-Time Polymerase Chain Reaction ; Repressor Proteins ; physiology ; Survival Analysis ; Uterine Cervical Neoplasms ; genetics ; mortality

OBJECTIVETo evaluate the roles of various cytokines, histone acetyltransferase (HAT) and histone deacetylase (HDAC) in the pathogenesis of bronchial asthma.

METHODSBALB/C mice were randomly assigned to control, untreated asthma, hormone treatment and TSA treatment groups. Bronchial asthma was induced by intraperitoneal injections and atomization inhalation of ovalbumin (OVA) in the asthma, hormone treatment and trichostatin (TSA) treatment groups. The mice in the hormone treatment and TSA treatment groups were administered with dexamethasone 1.0 mg/kg and TSA 1.0 mg/kg respectively by an intraperitoneal injection 30 minutes before challenge of asthma. At 24 hours after the last challenge, IL-4, IL-8 and IFN- levels in serum were measured using ELISA, and activities of HAT and HDAC in peripheral blood mononuclear cells (PBMC) were determined by the enzyme linked immunofluorescence assay.

RESULTSThe serum levels of IL-4 and IL-8 in the untreated asthma group were higher than in the control, hormone treatment and TSA treatment groups (P<0.05). There was no difference in the serum levels of IL-4 and IL-8 among the control, hormone treatment and TSA treatment groups (P>0.05). The activity of HDAC in the untreated asthma group was lower than in the control, hormone treatment and TSA treatment groups (P<0.05). Hormone treatment significantly decreased the activity of HAT compared with the untreated asthma group (P<0.05). There was no difference in the activities of HAT and HDAC among the control, hormone treatment and TSA treatment groups (P>0.05).

CONCLUSIONSThe decreased activity of HDAC leads to an increased secretion of inflammatory factors and thus induces asthma.

Animals ; Asthma ; enzymology ; etiology ; immunology ; Cytokines ; blood ; Histone Acetyltransferases ; physiology ; Histone Deacetylases ; physiology ; Male ; Mice ; Mice, Inbred BALB C ; Th1 Cells ; immunology ; Th2 Cells ; immunology

Histone deacetylases are involved in many biological processes and have roles in regulating cell behaviors such as cell cycle entry, cell proliferation and apoptosis. However, the effect of histone deacetylases on the development of hair cells (HCs) has not been fully elucidated. In this study, we examined the influence of histone deacetylases on the early development of neuromasts in the lateral line of zebrafish. Hair cell development was evaluated by fluorescent immunostaining in the absence or presence of histone deacetylase inhibitors. Our results suggested that pharmacological inhibition of histone deacetylases with inhibitors, including trichostatin A, valproic acid and MS-275, reduced the numbers of both HCs and supporting cells in neuromasts. We also found that the treatment of zebrafish larvae with inhibitors caused accumulation of histone acetylation and suppressed proliferation of neuromast cells. Real-time PCR results showed that the expression of both p21 and p27 mRNA was increased following trichostatin A treatment and the increase in p53 mRNA was modest under the same conditions. However, the expression of p53 mRNA was significantly increased by treatment with a high concentration of trichostatin A. A high concentration of trichostatin A also led to increased cell death in neuromasts as detected in a TUNEL assay. Moreover, the nuclei of most of these pyknotic cells were immunohistochemically positive for cleaved caspase-3. These results suggest that histone deacetylase activity is involved in lateral line development in the zebrafish and might have a role in neuromast formation by altering cell proliferation through the expression of cell cycle regulatory proteins.
Animals ; Apoptosis ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor Proteins/genetics/metabolism ; Histone Deacetylase Inhibitors/*pharmacology ; Histone Deacetylases/*metabolism ; Histones/metabolism ; Larva/growth & development/metabolism ; Lateral Line System/cytology/*growth & development/metabolism ; Mechanoreceptors/drug effects/*metabolism/physiology ; RNA, Messenger/genetics/metabolism ; Zebrafish ; Zebrafish Proteins/*metabolism

Acetylation ; Animals ; Child ; DNA Methylation ; Epigenesis, Genetic ; Heart ; physiology ; Heart Defects, Congenital ; etiology ; genetics ; metabolism ; Histone Acetyltransferases ; metabolism ; Histone Deacetylases ; metabolism ; Histones ; chemistry ; genetics ; physiology ; Humans ; Protein Processing, Post-Translational ; Transcription Factors ; metabolism

Angiogenesis, a process by which the preexisting blood vasculature gives rise to new capillary vessels, is associated with a variety of physiologic and pathologic conditions. However, the molecular mechanism underlying this important process remains poorly understood. Here we show that histone deacetylase 6 (HDAC6), a microtubule-associated enzyme critical for cell motility, contributes to angiogenesis by regulating the polarization and migration of vascular endothelial cells. Inhibition of HDAC6 activity impairs the formation of new blood vessels in chick embryos and in angioreactors implanted in mice. The requirement for HDAC6 in angiogenesis is corroborated in vitro by analysis of endothelial tube formation and capillary sprouting. Our data further show that HDAC6 stimulates membrane ruffling at the leading edge to promote cell polarization. In addition, microtubule end binding protein 1 (EB1) is important for HDAC6 to exert its activity towards the migration of endothelial cells and generation of capillary-like structures. These results thus identify HDAC6 as a novel player in the angiogenic process and offer novel insights into the molecular mechanism governing endothelial cell migration and angiogenesis.
Anilides ; pharmacology ; Animals ; Cell Movement ; Cell Polarity ; Cells, Cultured ; Chick Embryo ; Chickens ; Endothelial Cells ; cytology ; Histone Deacetylase 6 ; Histone Deacetylases ; metabolism ; physiology ; Humans ; Hydroxamic Acids ; pharmacology ; Mice ; Microtubule-Associated Proteins ; metabolism ; physiology ; Neovascularization, Physiologic

BACKGROUNDNF-kappaB p65 was shown to inhibit transcription of phosphoenolpyruvate carboxykinase (PEPCK), a rate-limiting enzyme in gluconeogenesis in the liver. To understand the mechanism of action of NF-kappaB p65, we investigated the nuclear receptor corepressor in the regulation of PEPCK transcription.

METHODSRat H4IIE cells, human hepatoma HepG2 cells and human embryo kidney (HEK) 293 cells were used in this study. The transcriptional activity of a rat PEPCK gene promoter (-490/+100) was analyzed in HepG2 cells, a HepG2 super suppressor IkBalpha (ssIkBalpha) stable cell line, and HEK 293 cells. The effects of p65 and ssIkBalpha on a rat PEPCK gene promoter were observed using the PEPCK luciferase reporter system. The interaction of the cAMP-response- element-binding (CREB) protein, histone deacetylase 3 (HDAC3) and silencing mediator for retinoic and thyroid hormone receptors (SMRT) with the PEPCK gene promoter were investigated using the chromatin immunoprecipitation (ChIP) assay. p65 cotransfection and RNAi-mediated gene knockdown were used to determine the corepressor involved in the inhibition of PEPCK by NF-kappaB p65 and the transcriptional regulation of CREB by NF-kappaB p65.

RESULTSNF-kappaB p65 inhibited PEPCK expression and the inhibition was blocked by ssIkBalpha. The inhibitory effect of p65 was completely blocked in a HepG2 stable cell line in which ssIkBalpha was expressed. HDAC3 or SMRT knockdown led to a significant up-regulation of PEPCK reporter activity in the presence of p65 cotransfection. In the ChIP assay the interaction of HDAC3 and SMRT with the PEPCK gene promoter was induced by p65 activation, but the CREB signal was reduced. Transcriptional activity of CREB was inhibited by NF-kappaB p65 cotransfection. The inhibitory effect of NF-kappaB p65 was blocked by HDAC3 RNAi or SMRT RNAi.

CONCLUSIONSThe study showed that the inhibition of PEPCK by NF-kappaB p65 was dependent on HDAC3 and SMRT, which form a nuclear corepressor complex for transcriptional inhibition. The transcription factors NF-kappaB p65 and CREB share the same corepressor HDAC3-SMRT, and the corepressor exchange leads to inhibition of PEPCK gene transcription by NF-kappaB p65.

Animals ; Blotting, Western ; Cell Line ; Chromatin Immunoprecipitation ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Hep G2 Cells ; Histone Deacetylases ; genetics ; metabolism ; Humans ; NF-kappa B ; genetics ; metabolism ; Nuclear Receptor Co-Repressor 2 ; genetics ; metabolism ; Phosphoenolpyruvate Carboxykinase (ATP) ; genetics ; Promoter Regions, Genetic ; genetics ; Protein Binding ; genetics ; physiology ; Rats ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factor RelA ; genetics ; metabolism

OBJECTIVETo investigate the inhibitory effect on MTA1 gene by MTA1 short hairpin RNA (shRNA), downstream regulation of expression of ERa and invasion of human breast cancer cells.

METHODSRecombinant plasmid pGenesil-1/MTA1 was constructed and transfected into human breast cancer cell lines MDA-MB-231 and MCF-7. Fluorescence microscopy was used to evaluate the efficacy of transfection. The transcription expression of MTA1 and MMP-9 gene were determined by reverse transcription-polymerase chain reaction (RT-PCR), the protein expression of ERalpha and MMP-9 were determined by immunohistochemical and western blot. The invasion ability was evaluated by Boyden chamber invasive assay.

RESULTSRecombinant plasmid pGenesil-1/MTA1 was constructed and transfected into MDA-MB-231 (82.5%) and MCF-7 (78.2%) successfully. After the transfection, MTA1 mRNA was suppressed in both cell lines (80.2% and 58.7%). The expression of ERalpha protein became positive in transfected MDA-MB-231 cell, and the expression of MMP-9 mRNA were down-regulated. The invasion ability was decreased (27.2% +/- 2.1)% compared with (76.3% +/- 2.4%), (P < 0.05). In contrast, transfected MCF-7 cells failed to show significant difference in the expression of ERalpha and MMP-9, without change of the invasion ability (P > 0.05).

CONCLUSIONSRNA interference can effectively suppress the expression of MTA1 in human breast cancer cells. An induction of ERalpha protein and suppression of MMP-9 may be related to the decrease of tumor cell invasive ability. RNA interference involving MTA1 gene may provide an effective anti-cancer gene therapy.

Blotting, Western ; Breast Neoplasms ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Down-Regulation ; drug effects ; Estrogen Receptor alpha ; genetics ; metabolism ; Histone Deacetylases ; genetics ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; RNA Interference ; physiology ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; pharmacology ; Repressor Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

AIMTo investigate the stage-specific localization of metastasis-associated protein 1 (MTA1) during spermatogenesis in adult human and mouse testis.

METHODSThe immunolocalization of MTA1 was studied by immunohistochemistry and Western blot analysis. The distribution pattern of MTA1 in mouse testis was confirmed by using quantitative analysis of purified spermatogenic cells.

RESULTSThe specificity of polyclonal antibody was confirmed by Western blot analysis. MTA1 was found expressed in the nucleus of germ cells, except elongate spermatids, and in the cytoplasm of Sertoli cells; Leydig cells did not show any specific reactivity. MTA1 possessed different distribution patterns in the two species: in humans, the most intensive staining was found in the nucleus of round spermatids and of primary spermatocytes while in mice, the most intense MTA1 staining was in the nucleus of leptotene, zygotene and pachytene spermatocytes. In both species the staining exhibited a cyclic pattern.

CONCLUSIONThe present communication initially provides new evidence for the potential role of MTA1 in mature testis. In addition, its distinctive expression in germ cells suggests a regulatory role of the peptide during spermatogenesis.

Adult ; Animals ; Animals, Outbred Strains ; Blotting, Western ; Histone Deacetylases ; metabolism ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Repressor Proteins ; metabolism ; Sexual Maturation ; physiology ; Species Specificity ; Spermatogenesis ; physiology ; Testis ; cytology ; metabolism ; Transcription Factors ; metabolism