scholarly journals Evaluation of Putative Type II Toxin-Antitoxin Systems and Lon Protease Expression in Shigella flexneri Following Infection of Caco-2 Cells

2020 ◽  
Vol 15 (3) ◽  
Author(s):  
Erfan Kheradmand ◽  
Shabnam Razavi ◽  
Malihe Talebi ◽  
Mahmood Jamshidian
Keyword(s):  
Cell Line ◽  
Post Infection ◽  
Shigella Flexneri ◽  
Fold Increase ◽  
Expression Level ◽  
Bacterial Invasion ◽  
P Value ◽  
Lon Protease ◽  
Expression Levels ◽  
Pcr Method

: Shigella flexneri causes bacillary dysentery in developing countries. Due to recent reports regarding antimicrobial resistance in human S. flexneri, finding alternative therapeutics is of vital importance. Toxin-antitoxin (TA) systems have recently been introduced as antimicrobial targets owing to their involvement in bacterial survival in stress conditions and “persister” cell formation. In this study, the presence of four TA loci were studied in S. flexneri ATCC 12022. The presence of genes coding for the identified TA loci and Lon protease were confirmed by the PCR method using specific primers. Caco-2 cell lines were then infected with this standard strain, and 8 and 24 h post-infection, expression levels of genes coding for the studied TA loci, and Lon protease were evaluated using a real-time PCR method. Expression of mazF, GNAT (Gcn5-related N-acetyltransferase), yeeU, pfam13975, and Lon genes showed 5.4, 9.8, 2.3, 2.7, and 13.8-fold increase, respectively, 8 h after bacterial invasion of the Caco-2 cell line. In addition, the expression of the aforementioned genes showed 4.8, 10.8, 2.3, 3.7, and 16.8-fold increase after 24 h. The GNAT and lon genes showed significantly higher expression levels compared to the control (P value < 0.05). However, the increase in the expression level of yeeU was the same at 8 h and 24 h post-infection. In addition, mazF expression level showed a slight decrease at 24 h compared to 8h post-infection. Genes coding for GNAT and Lon protease showed a significantly higher expression after invading the Caco-2 cell line. Therefore, targeting GNAT or Lon protease can be taken into consideration for finding novel antimicrobial drug strategies. The exact functions and mechanisms of TA systems in S. flexneri isolates are suggested to be experimentally determined.

scholarly journals Impact of either Elevated or Decreased Levels of Cytochrome bd Expression on Shigella flexneri Virulence

1999 ◽  
Vol 181 (4) ◽  
pp. 1229-1237 ◽  
Author(s):  
Sing Sing Way ◽  
Sandra Sallustio ◽  
Richard S. Magliozzo ◽  
Marcia B. Goldberg
Keyword(s):  
Shigella Flexneri ◽  
Lethal Dose ◽  
Fold Increase ◽  
Intracellular Survival ◽  
Bacterial Invasion ◽  
Wild Type ◽  
Terminal Oxidases ◽  
Cytochrome Bd ◽  
Transport Chain ◽  
Pathogenic Process

ABSTRACT Shigella spp. are the major cause of bacillary dysentery worldwide. The pathogenic process involves bacterial invasion and lysis of the phagocytic vacuole, followed by replication and movement within the cell cytoplasm and, ultimately, spread directly into adjacent cells. This study demonstrates that S. flexneri cytochrome bd expression is necessary for normal intracellular survival and virulence. Cytochrome bdis one of two terminal oxidases in the bacterial respiratory chain that reduce molecular oxygen to water, utilizing intermediates shuttled through the electron transport chain. S. flexneri mutants that contain a disruption in the cydC locus, which leads to defective cytochrome bd expression, or in the riboflavin (ribE) or ubiquinol-8 (ubiH) biosynthetic pathway, which leads to elevated cytochrome bd expression, were evaluated in intracellular survival and virulence assays. ThecydC mutant formed significantly smaller plaques, had significantly decreased intracellular survival, and had a 100-fold increase in lethal dose for mice compared with the wild type. TheribE and ubiH mutants each formed significantly larger plaques and had a 10-fold decrease in lethal dose for mice compared with the wild type. The data indicate that expression of cytochrome bd is required for S. flexneriintracellular survival and virulence.

scholarly journals Activation of JAK2/STAT5 Pathway Reduces Expression Level of DNMT3a in MPN Cell Line

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5394-5394
Author(s):  
Jie Zhou ◽  
Aibin Liang ◽  
Shaoguang Li ◽  
Wenjun Zhang ◽  
Jianfei FU
Keyword(s):  
Protein Expression ◽  
Cell Line ◽  
Dna Methyltransferase ◽  
De Novo ◽  
Conflicts Of Interest ◽  
Myeloproliferative Neoplasms ◽  
Expression Level ◽  
Hematopoietic Stem ◽  
Expression Levels ◽  
Increased Risk

Introduction: Myeloproliferative neoplasms (MPN) are clonal hematopoietic stem cell (HSC) disorders characterized by overproduction of mature blood cells and increased risk of transformation to acute myeloid leukemia (AML), and JAK2V167F is the most frequent MPN driving mutation detected in >95% of PV and 50-60% ET and PMF. DNMT3A is a de novo DNA methyltransferase that catalyzes the addition of methyl groups into active chromatin in CpG-rich regions leading to gene inactivation. Dnmt3a-/- HSC have enhanced self-renewal and a block in differentiation in vivo. Previous study showed that JAK2V617F and Dnmt3a loss cooperate to induce myelofibrosis through activated enhancer-driven inflammation, while whether JAK2V617F regulates DNMT3a still remains unclear. AZ960 is a potent and selective ATP competitive inhibitor of the JAK2 kinase, and previous studies reported that AZ960 possessed the activity selectively against JAK2. LY2784544 has been identified as a selective inhibitor of JAK2V617F and has undergone clinical trials for the treatment of several myeloproliferative disorders. Methods: Empty vector (control) and mutant JAK2V617F were transduced into BaF3 cells using a lentivirus system. JAK2V617F-expressing BaF3 cells grow IL-3 independent and were selected by fluorescence-activated cell sorting (FACS) for GFP expression. The protein expression levels of p-STAT5 and DNMT3a were detected by western blotting. JAK2V617F-expressing and control BaF3 cells were incubated with gradient concentration of LY2784544 or AZ960 to inhibit JAK2/STAT5 pathway. Results: The expression levels of p-STAT5 were obviously up-regulated in the JAK2V617F-expressing BaF3 cells, and DNMT3a was down-regulated. After 1-hour incubation in the serial diluted LY2784544, p-STAT5 were reduced in JAK2V617F-expressing BaF3 cells, with expression of DNMT3a elevated. To further confirm the correlation between JAK2/STAT5 pathway and expression of DNMT3a, another JAK2 inhibitor AZ960 was tested similar to LY2784544. With p-STAT5 expression suppressed, protein level of DNMT3a showed significantly promotion. Conclusion: We observed that JAK2V167F mutation suppresses protein expression levels of DNMT3a in MPN cell lines. JAK2 inhibition by AZ960 and LY2784544 significantly improved expression levels of DNMT3a. The activation of JAK2/STAT5 pathway reduces expression level of DNMT3a in MPN cell line, and the specific mechanism still needs to be explored. Figure Disclosures No relevant conflicts of interest to declare.

scholarly journals Hippocampal LMNA Gene Expression is Increased in Late-Stage Alzheimer’s Disease

2019 ◽  
Vol 20 (4) ◽  
pp. 878 ◽  
Author(s):  
Iván Méndez-López ◽  
Idoia Blanco-Luquin ◽  
Javier Sánchez-Ruiz de Gordoa ◽  
Amaya Urdánoz-Casado ◽  
Miren Roldán ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Late Stage ◽  
Fold Increase ◽  
Transcript Level ◽  
P Value ◽  
Mrna Levels ◽  
Lmna Gene ◽  
Expression Levels ◽  
Human Hippocampus

Lamins are fibrillary proteins that are crucial in maintaining nuclear shape and function. Recently, B-type lamin dysfunction has been linked to tauopathies. However, the role of A-type lamin in neurodegeneration is still obscure. Here, we examined A-type and B-type lamin expression levels by RT-qPCR in Alzheimer’s disease (AD) patients and controls in the hippocampus, the core of tau pathology in the brain. LMNA, LMNB1, and LMNB2 genes showed moderate mRNA levels in the human hippocampus with highest expression for the LMNA gene. Moreover, LMNA mRNA levels were increased at the late stage of AD (1.8-fold increase; p-value < 0.05). In addition, a moderate positive correlation was found between age and LMNA mRNA levels (Pearson’s r = 0.581, p-value = 0.018) within the control hippocampal samples that was not present in the hippocampal samples affected by AD. A-type and B-type lamin genes are expressed in the human hippocampus at the transcript level. LMNA mRNA levels are up-regulated in the hippocampal tissue in late stages of AD. The effect of age on increasing LMNA expression levels in control samples seems to be disrupted by the development of AD pathology.

scholarly journals Expression Changes of the FOXE1 and Lncrna PTCSC2 Expression in Tumor Tissues Compared with Normal Tissues in Patients with Colorectal Cancer

2022 ◽  
Author(s):  
Marzieh Ghani Dehkordi ◽  
Maryam Peymani
Keyword(s):  
Colorectal Cancer ◽  
Tumor Tissue ◽  
Expression Patterns ◽  
Study Groups ◽  
Expression Level ◽  
P Value ◽  
Expression Levels ◽  
Normal Tissues ◽  
Tumor Tissues ◽  
Significant Difference

Introduction: In recent studies, methylation of FOXE1 in colorectal cancer has been reported as a diagnostic biomarker. In this study for the first time, the expression of FOXE1 and PTCSC2 in colorectal cancer was investigated and their expression patterns in two healthy and tumor tissues of patients were compared. Methods: In this study, 40 tumor tissues with colorectal cancer and 40 adjacent normal samples were collected. Total RNA was extracted and cDNA synthesis followed. Then, the specific genes for lncRNA PTCSC2 and FOXE1 were amplified. The results were statistically analyzed by Graph Pad Prism software and a T-test was used to compare the expression levels of lncRNA PTCSC2 and FOXE1 in the patients and healthy group; p-value less than 0.05 was considered significant difference criteria. Results: In this study, the FOXE1 expression level was significantly decreased in tumor tissue (p-value = 0.005), whereas the lncRNA PTCSC2 expression level in tumor tissue was not significantly changed (p-value = 0.65). In addition, the expression levels of FOXE1 and lncRNA PTCSC2 did not show a significant relation with disease progression and age of the patients. ROC curve for changes in FOXE1 and lncRNA PTCSC2 expression showed that theFOXE1 gene could be a relatively appropriate independent variable (p-value = 0.03) to differentiate between the two study groups. Conclusion: According to the results of this study, changes in FOXE1 gene expression were significantly reduced in tumor samples and can be used as a biomarker in tumor diagnosis in colorectal cancer.

Knockdown Of The Piwi - Like Protein 4 (PIWIL4) Delays Leukemic Growth and Is Associated With Gross Changes In The Global Histone Methylation Marks In Human MLL - Rearranged AML

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 597-597 ◽  
Author(s):  
Shiva Bamezai ◽  
Medhanie M Mulaw ◽  
Fengbiao Zhou ◽  
Christian Rohde ◽  
Carsten Muller-Tidow ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Line ◽  
Conflicts Of Interest ◽  
Complex Function ◽  
Fold Increase ◽  
Differentially Expressed ◽  
Human Cord Blood ◽  
Hematopoietic Stem ◽  
Expression Levels

Abstract Piwi proteins belong to a class of proteins which were shown to be critically involved in the maintenance of the self-renewal property of stem cells in lower organisms. Furthermore, it was shown that they preserve genomic integrity through epigenetic silencing of transposable elements via CpG methylation and repressive histone modifications such as H3K9me3 in close interaction with a novel class of non-coding RNA called piRNA. So far there are neither precise data on the function of Piwi proteins in human acute myeloid leukemia, nor are there reports on expression of piRNAs in this disease. In a first step we tested PIWIL gene expression levels in normal human hematopoietic cells and leukemic patient samples by qRT-PCR. Among the family of human PIWI genes, PIWIL4 showed the highest expression level and was ubiquitously expressed in normal hematopoietic stem/progenitors, mature lymphoid and myeloid cells. Importantly, PIWIL4 showed aberrantly high expression in more than 72% of the AML patients (n=68; p< 0.0001) compared to normal CD34+ bone marrow (BM) and total BM cells (n=3). Notably, in nine of the ten MLL-AF9 rearranged AML patients, PIWIL4 was 64-fold higher expressed compared to normal CD34+ BM (p<0.0001) and 8-fold higher compared to inv(16), PML-RARa or cytogenetically normal AML patients (p<0.0001). To further validate this finding we analysed gene expression data performed on CD34+ human cord blood cells transduced with MLL-AF9 (n=9) vs AML-ETO (n=6) vs MYH11 (n=3): of note, PIWIL4 showed a 6 fold increase in expression in the MLL-AF9 transduced cells compared to the other experimental arms. Stable knockdown of PIWIL4 in the MLL rearranged AML cell lines MV4-11 (MLL-AF4) and THP-1 (MLL-AF9) significantly impaired growth in vitro (n=3) reducing proliferation and clonogenic growth by 83%/93% and 91%/93%, respectively. In addition, depletion of PIWIL4 delayed onset of leukemia in NSG mice transplanted with MV4-11/ THP-1 cells transduced with shPIWIL4 compared to the scrambled control (shRNA: AML onset 48/62d after transplantation vs. 30/30 days in the scrambled control; n=4/8 per arm; p< 0.0001/p<0.001). ChIP-seq analysis revealed that depletion of PIWIL4 in the THP1 cell line results in a marked global reduction in repressive H3K9me3 marks and in an increase in activating H3K4me3 marks as compared to cells transduced with the scrambled control. RNA-seq analyses revealed over 2500 differentially expressed genes upon PIWIL4 depletion with 60% of the genes being upregulated compared to the scrambled control (p<0.05). Among them genes involved in cell cycle such as RB1, P21, TGFB1 as well as epigenetic modifiers such as SETDB1, HDAC1,2 and demethylating enzyme TDG were differentially expressed. RB1 and EED, a protein necessary for PRC2 complex function, displayed an increase in expression and loss of H3K9me3 modifications on their promoters upon knockdown of PIWIL4. To prove piRNA expression in human AML and to test any association between PIWIL4 expression and piRNA signatures, microarray analyses covering 23,677 piRNAs was performed on the MLL-AF9 rearranged THP-1 cell line, of which 14193 piRNAs showed expression levels higher than 4 (arbitrary log2 scale). PIWIL4 knockdown induced differential expression of 981 piRNAs (p≤0.01, fold change ≥2), of which 527 were downregulated and 454 upregulated. Thus, collectively, we could show for the first time that PIWIL4 expression is deregulated in human AML, affects leukemic growth, shapes epigenetic marks and impacts piRNA expression in this disease. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Evaluation of expression of VDR-associated lncRNAs in COVID-19 patients

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Mohammad Taheri ◽  
Lina Moallemi Rad ◽  
Bashdar Mahmud Hussen ◽  
Fwad Nicknafs ◽  
Arezou Sayad ◽  
...  
Keyword(s):  
Healthy Subjects ◽  
Distress Syndrome ◽  
Current Data ◽  
Hospitalized Patients ◽  
P Value ◽  
Healthy Controls ◽  
Expression Levels ◽  
Significant Difference ◽  
Male Patients ◽  
Pcr Method

Abstract Background Coronavirus disease 2019 (COVID-19) has been shown to cause serious health problems among them is the Acute Respiratory Distress syndrome (ARDS). Vitamin D receptor (VDR) signaling possibly partakes in the pathophysiology of this devastating complication. Methods In the current project, we have appraised expression levels of VDR, CYP27B1 and a number of associated lncRNAs in the circulation of COVID-19 patients versus healthy subjects using real-time PCR method. Results Expression of SNHG6 was considerably lower in COVID-19 patients compared with control subjects (Ratio of mean expression (RME) = 0.22, P value = 7.04E-05) and in both female and male COVID-19 patients compared with sex-matched unaffected individuals (RME = 0.32, P value = 0.04 and RME = 0.16, P value = 0.000679683, respectively). However, its expression was similar among ICU-hospitalized and non-ICU patients. Similarly, expression of SNHG16 was lower in in COVID-19 patients compared with controls (RME = 0.20, P value = 5.94E-05) and in both female and male patients compared with sex-matched controls (RME = 0.32, P value = 0.04 and RME = 0.14, P value = 0.000496435, respectively) with no significant difference among ICU-hospitalized and non-ICU hospitalized patients. Expression of VDR was lower in COVID-19 patients compared with controls (RME = 0.42, P value = 0.04) and in male patients compared with male controls (RME = 0.27, P value = 0.02). Yet, expression of VDR was statistically similar between female subgroups and between ICU-hospitalized and non-ICU hospitalized patients. Expression levels CYP27B, Linc00511 and Linc00346 were similar among COVID-19 patients and healthy subjects or between their subgroups. Significant correlations have been detected between expression levels of VDR, CYP27B and SNHG6, SNHG16, Linc00511 and Linc00346 lncRNAs both among COVID-19 patients and among healthy controls with the most significant ones being SNHG6 and SNHG16 (r = 0.74, P value = 3.26e-17 and r = 0.81, P = 1.54e-22, respectively). Conclusion Combination of transcript levels of VDR, CYP27B and SNHG6, SNHG16, Linc00511 and Linc00346 could differentiate patients from controls with AUC = 0.76, sensitivity = 0.62 and specificity = 0.81. The current data potentiate SNHG6, SNHG16 and VDR as possible contributors in COVID-19 infection but not in the severity of ARDS.

scholarly journals The Changes of Expression Levels in Mir-181a and Mir-30d, and a Significant Correlation between Clinical Patient Data

2019 ◽  
Vol 8 (1) ◽  
pp. 16
Author(s):  
Mojtaba MohammadnejhadMohammadnejad Pahmadani ◽  
Fatemeh Jabari ◽  
Shima Hojabri Mahani ◽  
Reza Mahmanzar
Keyword(s):  
Colorectal Cancer ◽  
Rna Extraction ◽  
Cancer Biomarkers ◽  
Expression Level ◽  
Expression Levels ◽  
Clinical Patient ◽  
Tumor Tissues ◽  
Significant Difference ◽  
Pcr Method ◽  
Tumor Expression

Purpose: Colorectal cancer is known as the most common gastrointestinal cancers. As the age increases, the risk for this cancer also increases, so the only way to improve and hope for life in these patients is early diagnosis of the disease. So far, numerous molecular studies have been carried out on microRNAs in colorectal cancer. In addition, since some of them can be identified as cancer biomarkers. Therefore, in this study we have investigated the expression level of Mir-30d and Mir-181a as cancer biomarkers. Method: The changes of Mir-30d and Mir-181a expression levels in 60 colorectal tumor tissues and 60 adjacent tumor tissues, after RNA extraction and cDNA synthesis were surveyed using the Real Time-PCR method. Results: The results have reported a considerable reduction in the expression level of Mir-30d in tumor tissues, as well as a significant increase in the expression level of Mir-181a tumor expression in tumor tissues (P&lt;0.05). In addition, the correlation between Mir-30d and Mir181a showed that there was a significant difference between the level of expression of mir-30d with age and TNM stage of CRC (P&lt;0.05), whilst these correlations were not observed for Mir-181a (P&gt;0.05).

scholarly journals Increased expression level of ANGPTL8 in White Adipose tissue under Chronic Cold Treatment

2021 ◽  
Author(s):  
Hossein Arefanian ◽  
Irina Al-Khairi ◽  
Nermeen Abu Khalaf ◽  
Preethi Cherian ◽  
Sina Kavalakatt ◽  
...  
Keyword(s):  
Gene Expression ◽  
Adipose Tissue ◽  
White Adipose Tissue ◽  
Energy Homeostasis ◽  
Cold Treatment ◽  
Fold Increase ◽  
Gene Expression Level ◽  
Expression Level ◽  
P Value ◽  
Brown Adipose

Abstract Background It is well known that Angiopoietin like protein (ANGPTL) mainly 3, 4 and 8 play a major role in maintaining energy homeostasis by regulating lipoprotein lipase (LPL) activity, which is a key regulator of triglyceride (TG) metabolism. Our aim is to evaluate the level of ANGPTL3, 4 and 8 in mice maintained under cold conditions in the liver, brown adipose tissue (BAT), and white adipose tissue (WAT). Methods C57BL/6J mice were exposed to cold temperature at 4˚C over a period of ten days with food given ad libitum. Animal tissues were harvested at days 0, 1, 3, 5, and 10 (cold treatment groups, n = 10 in each group, control, n = 5). Liver, subcutaneous adipose tissue (SAT), and BAT were used to investigate the expression level of different genes. ANGPTL3, 4 and 8 genes expression were measured in liver. ANGPTL4, 8 and UCP1 were measured in SAT and BAT. Results ANGPTL 3 and 8 gene expression levels were significantly reduced in mice liver tissues after cold treatment (P-value < 0.05). However, the gene expression level of ANGPTL4 was not significantly changed. In BAT, ANGPTL8 expression was not changed after cold treatment while ANGPTL4 was significantly reduced (P-value < 0.05). ANGPTL4 level was also significantly reduced in SAT, whereas the gene expression level of ANGPTL8 showed over a 5-fold increase. Similarly, the UCP1 gene expression was also significantly increased in SAT. Additionally, protein overexpression of ANGPTL8 was further confirmed by immunohistochemistry after extended cold treatment. Conclusion Our data shows that ANGPTL proteins are inhibited in the liver and BAT under cold treatment. This agrees with other studies that showed that reduction in ANGPTL4 in BAT improved thermogenesis in response to acute cold exposure. However, in our study we also observed that ANGPTL8 is activated under these conditions in SAT. This suggests that it might be involved in the regulation of lipolysis as well as enhancing SAT browning.

scholarly journals The Effect of Epigallocatechin gallate on Cross-talk Between Autophagy and Apoptosis in NALM-6 Cell Line

2021 ◽  
Author(s):  
Faezeh Gharehchahi ◽  
Farahnaz Zare ◽  
Gholamreza Rafie Dehbidi ◽  
Zahra Yousefi ◽  
Somayeh Pourpirali ◽  
...  
Keyword(s):  
Cell Viability ◽  
Mrna Expression ◽  
Cell Line ◽  
Lymphoblastic Leukemia ◽  
Epigallocatechin Gallate ◽  
Fold Increase ◽  
Expression Level ◽  
Mrna Expression Level ◽  
Trypan Blue Exclusion ◽  
The Impact

Abstract Background: Acute lymphoblastic leukemia is a prevalent hematological malignancy in 2-5-year-old children. Chemotherapy, as the most common treatment for ALL, is not usually responsive. Epigallocatechin gallate (EGCG), a small molecule extracted from green tea, has significant effects on tumor cells through different mechanisms, such as DNA damage, cell cycle arrest, oxidative stress, apoptosis, and autophagy. In this study, we investigated the impact of EGCG on autophagy and apoptosis in NALM-6 cell line. Methods and results: Cell viability and apoptosis were assessed by MTT and Trypan blue exclusion assay, and flow cytometry. It was shown that EGCG remarkably inhibited proliferation, reduced cell viability, and induced apoptosis in NALM-6 cell line (P<0.05). In addition, real-time PCR and western blot analysis were used to examine autophagy. It was observed that EGCG resulted in a 4-fold increase in LC3 protein level (P<0.05) while reducing the mRNA expression level of LC3B, P62/SQSTM1, and Atg2B genes (P<0.01). It also caused around 1.3-fold increase in DRAM1 mRNA expression level (P<0.05). Finally, it was indicated that the inhibition of autophagy affects apoptosis neither in untreated nor treated cells with EGCG.Conclusion: These results show that EGCG can induce apoptosis and autophagy in NALM-6 cell line while inhibition of autophagy cannot affect apoptosis in this cell line.

Electroacupuncture ameliorates CUMS-induced depression-like behavior: involvement of the glutamatergic system and apoptosis in rats

2020 ◽  
Vol 23 ◽  
Author(s):  
Qin Guo ◽  
Xian-Ming Lin ◽  
Zhong Di ◽  
Quan-Ai Zhang ◽  
Shuo Jiang
Keyword(s):  
Nmda Receptor ◽  
Caspase 3 ◽  
B Cell Lymphoma ◽  
Sucrose Preference ◽  
Expression Level ◽  
Receptor Subunit ◽  
Glutamatergic System ◽  
Ionotropic Receptor ◽  
Nmda Receptor Subunit ◽  
Expression Levels

Background: Converging evidence indicates that glutamatergic system and glia are directly implicated in the pathophysiology of depression. Clinical studies indicate that electroacupuncture (EA) has antidepressant-like effect with low side effects for depression. However, the underlying antidepressant mechanism of acupuncture remains obscure. Methods: Chronic unpredictable mild stress (CUMS)-induced depressive rats were used to induce depressive-like behavior, and evaluated by the weight change, open field test, sucrose preference test, and novelty suppressed feeding test. EA, NMDA receptor subunit 2A antagonist (NR2A RA) or NMDA receptor subunit 2B antagonist (NR2B RA) was used for comparison. High performance liquid chromatography (HPLC) was performed to detect the content of hippocampal glutamate, while western blot for the hippocampal protein expression levels of calcium/calmodulin-dependent protein kinase II (CaMKII), Bax, caspase 3 and B-cell lymphoma-2 (Bcl-2). The distribution of glutamate ionotropic receptor NMDA type subunit 2A (NR2A), neuronal nuclear protein (NeuN), glutamate ionotropic receptor NMDA type subunit 2B (NR2B) and glial fibrillary acidic protein (GFAP) were detected by immunofluorescence. Results: Significant depression behavior (reduced body weight and sucrose preference, increased feeding and immobility time) was produced in CUMS-induced depressive rats, which was reversed significantly by EA. EA decreased hippocampal glutamate level. EA led to a significant decrease in expression levels of Bax, caspase 3 and CaMKⅡ accompanied by increased Bcl-2 expression level. Furthermore, EA significantly increased NR2A expression level as well as decreased NR2B expression level in hippocampus. Conclusion: EA ameliorated depression-like behavior in CUMS rats, which might be mediated, at least in part, by regulating the glutamate, NMDA receptors and apoptosis in the hippocampus.

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