scholarly journals Transient Focal Cerebral Ischemia Leads to miRNA Alterations in Different Brain Regions, Blood Serum, Liver, and Spleen

2021 ◽  
Vol 23 (1) ◽  
pp. 161
Author(s):  
Clara Voelz ◽  
Nahal Ebrahimy ◽  
Weiyi Zhao ◽  
Pardes Habib ◽  
Adib Zendedel ◽  
...  
Keyword(s):  
Cell Death ◽  
Cerebral Cortex ◽  
Ischemic Stroke ◽  
Blood Serum ◽  
Expression Profiles ◽  
Oxygen Deficiency ◽  
Brain Regions ◽  
Dependent Manner ◽  
Expression Levels ◽  
Infarct Area

Ischemic stroke is characterized by an occlusion of a cerebral blood vessel resulting in neuronal cell death due to nutritional and oxygen deficiency. Additionally, post-ischemic cell death is augmented after reperfusion. These events are paralleled by dysregulated miRNA expression profiles in the peri-infarct area. Understanding the underlying molecular mechanism in the peri-infarct region is crucial for developing promising therapeutics. Utilizing a tMCAo (transient Middle Cerebral Artery occlusion) model in rats, we studied the expression levels of the miRNAs (miR) 223-3p, 155-5p, 3473, and 448-5p in the cortex, amygdala, thalamus, and hippocampus of both the ipsi- and contralateral hemispheres. Additionally, the levels in the blood serum, spleen, and liver and the expression of their target genes, namely, Nlrp3, Socs1, Socs3, and Vegfa, were assessed. We observed an increase in all miRNAs on the ipsilateral side of the cerebral cortex in a time-dependent manner and increased miRNAs levels (miR-223-3p, miR-3473, and miR-448-5p) in the contralateral hemisphere after 72 h. Besides the cerebral cortex, the amygdala presented increased expression levels, whereas the thalamus and hippocampus showed no alterations. Different levels of the investigated miRNAs were detected in blood serum, liver, and spleen. The gene targets were altered not only in the peri-infarct area of the cortex but selectively increased in the investigated non-affected brain regions along with the spleen and liver during the reperfusion time up to 72 h. Our results suggest a supra-regional influence of miRNAs following ischemic stroke, which should be studied to further identify whether miRNAs are transported or locally upregulated.

scholarly journals Extracellular ASC exacerbated the recurrent ischemic stroke in an NLRP3-dependent manner

2019 ◽  
Vol 40 (5) ◽  
pp. 1048-1060 ◽  
Author(s):  
Xiao-fei He ◽  
Yi-xuan Zeng ◽  
Ge Li ◽  
Yu-kun Feng ◽  
Cheng Wu ◽  
...  
Keyword(s):  
Ischemic Stroke ◽  
Mouse Model ◽  
Inflammatory Responses ◽  
Recurrent Stroke ◽  
Nuclear Factor Κb ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Wild Type ◽  
Infarct Area ◽  
Recurrent Strokes

Using a photothrombotic mouse model of single stroke, we show that a single stroke onset increases the nuclear factor-κB (NF-κB), NLR family CARD domain containing protein 4 (NLRC4), and absent in melanoma 2 (AIM2) inflammasomes, as well as the mRNA levels of NLRP3. Next, using a photothrombotic mouse model of recurrent stroke, we found that recurrent strokes increased the activation of NLRP3, exacerbated the brain damage and the pro-inflammatory response in wild type (WT) mice, but not in NLRP3 knockout ( NLRP3 KO) mice. Additionally, we found that apoptosis-associated speck-like protein containing a CARD (ASC) protein level surrounding the infarct area was comparatively increased, but that ASC specks outside of microglia in both the ipsilateral and contralateral of stroke site were decreased in NLRP3 KO mice relative to wild-type (WT) controls, and the number of ASC specks surrounding the second infarct area was positively correlated to the damage scores. Mechanistically, we found that recombinant ASC (RecASC) activated NLRP3 and induced pro-inflammatory responses, exacerbating the outcome of ischemic stroke, in WT mice, but not in NLRP3 KO mice. We therefore conclude that the NLRP3 inflammasome is activated by two attacks of stroke, which act together with ASC to exacerbate recurrent strokes.

The IGF1-R Inhibitor AEW541 Is a Potent Anti-Myeloma Agent That Overcomes Drugs Resistance, and Has Synergistic Activity with Other New Drugs.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 2599-2599
Author(s):  
Patricia Maiso ◽  
Enrique M. Ocio ◽  
Mercedes Garayoa ◽  
Mark A. Pearson ◽  
Atanasio Pandiella ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Death ◽  
Expression Profiles ◽  
Chemotherapeutic Agents ◽  
Mitochondrial Outer Membrane ◽  
Late Time ◽  
Synergistic Activity ◽  
Dependent Manner ◽  
Myeloma Cells

Abstract Multiple myeloma (MM) represents an incurable disease for which development of new therapies is required. Here we report the effect on myeloma cells of AEW541, a new small molecule, belonging to the pyrrolo[2,3-d] pyrimidine class, identified as inhibitor of the IGF-1R in vitro kinase activity. AEW541 showed a potent antimyeloma activity (IC50 <4.5 μM) on MM cell lines both sensitive (MM1S, U266, OPM2, RPMI8226) and resistant (MM1R and U266LR7) to conventional chemotherapeutic agents. In fresh cells from five MM patients a marked antitumor activity was confirmed. AEW541 showed a synergistic effect with dexamethasone and lenalidomide, while it was additive with melphalan and bortezomib. Moreover the triple combination of AEW541, bortezomib and dexamethasone showed even higher anti-MM activity. Gene expression profiles of MM1S cells identified a total of 967 genes to be significantly deregulated (transcriptional changes in gene expression of 2-fold or greater) by treatment with AEW541. The classification of these genes according to functional categories indicated that 3.5% were involved in apoptosis/responses to stress and 13% in the control of cell cycle/proliferation. By Western analyses, we observed that AEW541 affected genes involved in cell cycle and cell death pathways. AEW541 blocked cell cycle progression, and this was accompanied by p27, up-regulation and pRb, CCND1, CCNA and CCNE downregulation. AEW541 induced cell death through an increase in the mitochondrial outer membrane permeability and provoked DNA fragmentation. AEW541 induced apoptosis and at late time points, also activated caspases 8,9 and 3. The pan-caspase inhibitor Z-VAD-FMK only slightly decreased the sensitivity to AEW541. In addition, AEW541 stimulated a caspase-independent pathway, through the release of AIF and Endonuclease G from the mitochondria. It is therefore conceivable that both caspase dependent and independent pathways are activated by AEW541 in MM cells, although the effect of AEW541 on cell cycle arrest is an earlier and more potent event. Finally, AEW541 was able to overcome the protective effect that confers IL-6, IGF-1 and BMSCs to myeloma cells in a dose dependent manner. All these data indicate that AEW541 could be a useful drug for the treatment of MM patients, particularly in combination with other novel agents such as bortezomib or lenalidomide together with dexamethasone.

scholarly journals Gamma-Aminobutyric Acid Increases Erythropoietin by Activation of Citrate Cycle and Stimulation of Hypoxia-Inducible Factors Expression in Rats

Biomolecules ◽  
2020 ◽  
Vol 10 (4) ◽  
pp. 595
Author(s):  
Keun-Tae Park ◽  
Jong-Kwon Han ◽  
Seong Jin Kim ◽  
Young-Hee Lim
Keyword(s):  
Nadh Dehydrogenase ◽  
Tricarboxylic Acid Cycle ◽  
Expression Profiles ◽  
Kidney Tissue ◽  
Physiological Role ◽  
Aminobutyric Acid ◽  
Dependent Manner ◽  
Gamma Aminobutyric Acid ◽  
Expression Levels ◽  
Gene And Protein Expression

Erythropoietin (EPO) is the primary regulator of erythropoiesis in the mammalian fetus and adult. Deficiency of EPO induces anemia. In this study, we investigated the effect of gamma-aminobutyric acid (GABA) on serum EPO levels and erythropoiesis in rats. Expression levels of Epo-related genes were measured by quantitative real-time PCR (qPCR) and expression of Epo and Epo receptor (Epor) proteins were measured by immunohistochemistry. The gene and protein expression profiles of kidney tissue in GABA-treated rats were evaluated by ribonucleic acid (RNA) sequencing and two-dimensional electrophoresis (2-DE), respectively. GABA significantly increased serum EPO levels and expression levels of Epo and Epor. GABA increased expression levels of hypoxia-inducible factor (Hif)-1 and Hif-2. Seven proteins with expression levels showing >2-fold change were identified by 2-DE followed by MALDI-TOF MS in GABA-treated rat kidney. The top KEGG pathway from the identified proteins was the tricarboxylic acid cycle, and nicotinamide adenine dinucleotide (NADH) dehydrogenase, succinate dehydrogenase, and isocitrate dehydrogenase were identified as key proteins. GABA treatment significantly increased ATP levels and NADH dehydrogenase activity in a dose-dependent manner. In conclusion, GABA shows a new physiological role in EPO production, and it can thus can contribute to the prevention of anemia when used alone or in combination with other anemia treating drugs.

scholarly journals Brief Focal Cerebral Ischemia That Simulates Transient Ischemic Attacks in Humans Regulates Gene Expression in Rat Peripheral Blood

2009 ◽  
Vol 30 (1) ◽  
pp. 110-118 ◽  
Author(s):  
Xinhua Zhan ◽  
Bradley P Ander ◽  
Glen Jickling ◽  
Renée Turner ◽  
Boryana Stamova ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Death ◽  
Ischemic Stroke ◽  
Cerebral Ischemia ◽  
Peripheral Blood ◽  
Focal Cerebral Ischemia ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Focal Ischemia ◽  
Transient Ischemic Attacks

Blood gene expression profiles of very brief (5 and 10 mins) focal ischemia that simulates transient ischemic attacks in humans were compared with ischemic stroke (120 mins focal ischemia), sham, and naïve controls. The number of significantly regulated genes after 5 and 10 mins of cerebral ischemia was 39 and 160, respectively (fold change ⩾∣1.5∣ and P<0.05). There were 103 genes common to brief focal ischemia and ischemic stroke. Ingenuity pathway analysis showed that genes regulated in the 5 mins group were mainly involved in small molecule biochemistry. Genes regulated in the 10 mins group were involved in cell death, development, growth, and proliferation. Such genes were also regulated in the ischemic stroke group. Genes common to ischemia were involved in the inflammatory response, immune response, and cell death—indicating that these pathways are a feature of focal ischemia, regardless of the duration. These results provide evidence that brief focal ischemia differentially regulates gene expression in the peripheral blood in a manner that could distinguish brief focal ischemia from ischemic stroke and controls in rats. We postulate that this will also occur in humans.

scholarly journals Molecular basis of the anti-cancer effects of genistein isoflavone in LNCaP prostate cancer cells

2011 ◽  
Vol 1 (3) ◽  
pp. 91 ◽  
Author(s):  
K. Merchant ◽  
J. Kumi-Diaka ◽  
A. Rathinavelu ◽  
N. Esiobu ◽  
R. Zoeller ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Cycle ◽  
Cell Death ◽  
Side Effects ◽  
Cancer Cells ◽  
The United States ◽  
Dependent Manner ◽  
Apoptosis Induction ◽  
Prostate Cancer Cells ◽  
Expression Levels

Background: Prostate cancer is the most common form of non-skin cancer within the United States and the second leading cause of cancer deaths. Survival rates for the advanced disease remain relatively low, and conventional treatments may be accompanied by significant side effects. As a result, current research is aimed at alternative or adjuvant treatments that will target components of the signal transduction, cell-cycle and apoptosis pathways, to induce cell death with little or no toxic side effects to the patient. In this study, we investigated the effect of genistein isoflavone, a soy derivative, on expression levels of genes involved in these pathways. The mechanism of genistein-induced cell death was also investigated. The chemosensitivity of the LNCaP prostate cancer cells to genistein was investigated using ATP and MTS assays, and a caspase binding assay was used to determine apoptosis induction. Several molecular targets were determined using cDNA microarray and RT-PCR analysis.Results: The overall data revealed that genistein induces cell death in a time- and dose-dependent manner, and regulates expression levels of several genes involved in carcinogenesis and immunity. Several cell-cycle genes were down-regulated, including the mitotic kinesins, cyclins and cyclin-dependent kinases. Various members of the Bcl-2 family of apoptotic proteins were also affected. The DefB1 and the HLA membrane receptor genes involved in immunogenicity were also up-regulated. Conclusion: The results indicate that genistein inhibits growth of the hormone-dependent prostate cancer cells, LNCaP, via apoptosis induction through regulation of some of the genes involved in carcinogenesis of many tumors, and immunogenicity. This study augments the potential phytotherapeutic and immunotherapeutic significance of genistein isoflavone. Key words: Genistein isoflavone, prostate cancer, expression of genes, phytotherapeutic adjuvant, immunotherapy and chemotherapy

C-fos, ERK1/2, MAP2, NOTCH1 Proteins Expression Patterns in Human Cerebral Cortex Neurons after Ischemic Stroke

2020 ◽  
Vol 75 (3) ◽  
pp. 226-233
Author(s):  
Svetlana P. Sergeeva ◽  
Aleksey V. Lyundup ◽  
Valery V. Beregovykh ◽  
Petr F. Litvitskiy ◽  
Aleksey A. Savin ◽  
...  
Keyword(s):  
Cerebral Cortex ◽  
Ischemic Stroke ◽  
Expression Patterns ◽  
Immunohistochemical Method ◽  
Left Middle Cerebral Artery ◽  
Contralateral Hemisphere ◽  
Human Cerebral Cortex ◽  
Fos Protein ◽  
Urgent Task ◽  
Indirect Immunoperoxidase

Background. The search for protein (these include c-fos, ERK1/2, MAP2, NOTCH1) expression that provide neuroplasticity mechanisms of the cerebral cortex after ischemic stroke (IS) patterns is an urgent task. Aims to reveal c-fos, ERK1/2, MAP2, NOTCH1 proteins expression patterns in human cerebral cortex neurons after IS. Materials and methods. We studied 9 left middle cerebral artery (LMCA) IS patients cerebral cortex samples from 3 zones: 1 the zone adjacent to the necrotic tissue focus; 2 zone remote from the previous one by 47 cm; 3 zone of the contralateral hemisphere, symmetric to the IS focus. Control samples were obtained from 3 accident died people. Identification of targeted proteins NSE, c-fos, ERK1/2, MAP2, NOTCH1 was performed by indirect immunoperoxidase immunohistochemical method. Results. Moving away from the ischemic focus, there is an increase in the density of neurons and a decrease in the damaged neurons proportion, the largest share of c-fos protein positive neurons in zone 2, NOTCH1 positive neurons in zone 1, smaller fractions of ERK1/2 and MAP2 positive neurons compared to the control only in samples of zone 1. Conclusions. With the IS development, the contralateral hemisphere is intact tissue increased activation zone, while the zones 1 and 2 have pathological activation signs. In zone 1 of the range, the adaptive response of the tissue decreases, and in zone 2 it expands. Therefore, a key target for therapeutic intervention is zone 2.

New aspects of the energy-tropic action of mexidol

2018 ◽  
pp. 36-40
Author(s):  
Ю.И. Кирова ◽  
Э.Л. Германова
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Cerebral Cortex ◽  
Growth Factor ◽  
Atp Synthase ◽  
Oxygen Deficiency ◽  
Vascular Endothelial ◽  
Cyt B ◽  
Respiratory Enzymes ◽  
Catalytic Subunits ◽  
Endothelial Growth Factor

Митохондриогенез и ангиогенез являются ключевыми нейропротекторными механизмами, повышающими устойчивость нервной ткани к условиям гипоксии/ишемии. В настоящее время фармакологическая индукция биогенеза митохондрий является одним из наиболее перспективных и активно разрабатываемых подходов к коррекции ишемических и постишемических нарушений, нейродегенеративных заболеваний и кардиопатий. Выявление факта стимуляции митохондриогенеза эталонными нейропротекторными препаратами позволит существенно расширить представление об их терапевтическом потенциале и принципах применения. Цель исследования - изучение влияния нейропротекторного сукцинатсодержащего препарата мексидол на экспрессию каталитических субъединиц дыхательных ферментов митохондрий, АТФ-синтазы и фактора роста эндотелия сосудов в коре головного мозга крыс с врожденными различиями в устойчивости к дефициту кислорода. Методика. Исследование выполнено на белых беспородных крысах-самцах. Инъекции мексидола (40 мг/кг, внутрибрюшинно) выполняли ежедневно на протяжении 20 сут. Уровень экспрессии каталитических субъединиц дыхательных ферментов митохондрий и АТФ-синтазы, фактора роста эндотелия сосудов и сукцинатного рецептора в ткани коры головного мозга оценивали методом иммуноблоттинга. Общую резистентность организма к острой гипоксии тестировали в гипобарической камере проточного типа при разрежении атмосферы, соответствующем 190 мм рт. ст. (3% О). Результаты. В ходе курса применения мексидола происходило увеличение уровня каталитических субъединиц дыхательных ферментов митохондрий (NDUFV2, SDHA, cyt b, COX1), АТФ-синтазы (ATP5A), фактора роста эндотелия сосудов (VEGF) и сукцинатного рецептора (SUCNR1), особенно выраженное у неустойчивых к гипоксии особей. Заключение. Впервые показана вовлеченность сукцинатсодержащего препарата мексидол в механизмы индукции ферментов энергопродуцирующей системы митохондрий коры головного мозга, что существенно расширяет сложившиеся представления о механизмах его энерготропного действия. Mitochondriogenesis and angiogenesis are crucial neuroprotective mechanisms that increase the resistance of nervous tissue to hypoxia/ischemia conditions. Currently, pharmacological induction of mitochondrial biogenesis is one of the most promising and actively developed approaches for the correction of ischemic and post-ischemic disorders, neurodegenerative diseases and cardiopathies. Revealing the fact of stimulation of mitochondriogenesis with standard neuroprotective drugs will significantly expand the understanding of their therapeutic potential and principles of application. The aim of the research was to study the effect of the neuroprotective succinate-containing drug mexidol on the expression of catalytic subunits of the respiratory enzymes of mitochondria, ATP-synthase and vascular endothelial growth factor in the cerebral cortex of rats with inborn differences in resistance to oxygen deficiency. Methods. The study was carried out on white mongrel rats-males injections of mexidol (40 mg/kg, intraperitoneally) were performed daily for 20 days. The expression level of the catalytic subunits of the respiratory enzymes of mitochondria and ATP-synthase, vascular endothelial growth factor and succinate receptor in the tissue of the cerebral cortex was assessed by immunoblotting. The total resistance of the organism to acute hypoxia was tested in a hypobaric chamber with an atmosphere underpressure corresponding to 190 mm Hg (3% O). Results. During the injection course of mexidol, there was an increase in the level of catalytic subunits of the respiratory enzymes of mitochondria (NDUFV2, SDHA, cyt b, COX1), ATP-synthase (ATP5A), vascular endothelial growth factor (VEGF) and succinate receptor (SUCNR1), especially pronounced in rats with low-resistance to hypoxia. Conclusion. The study revealed for the first time the involvement of the succinate-containing drug mexidol in the mechanisms of induction of enzymes of the energy-producing system of the mitochondria of the cerebral cortex, which significantly expands the existing ideas about the mechanisms of its energy-tropic action.

Irisin Ameliorates Hypoxia/Reoxygenation-Induced Inflammation and Apoptosis in PC12 Cells by Inhibiting TLR4/MYD88 Signaling Pathway

2019 ◽  
Vol 17 (3) ◽  
pp. 329-336
Author(s):  
Wang Jinli ◽  
Xu Fenfen ◽  
Zheng Yuan ◽  
Cheng Xu ◽  
Zhang Piaopiao ◽  
...  
Keyword(s):  
Ischemic Stroke ◽  
Signaling Pathway ◽  
Pc12 Cells ◽  
Major Complication ◽  
Lactic Dehydrogenase ◽  
Dependent Manner ◽  
Cerebral Ischemic Stroke ◽  
Cerebral Ischemic ◽  
Myd88 Signaling ◽  
Hypoxia Reoxygenation

Cardiovascular disease including cerebral ischemic stroke is the major complication that increases the morbidity and mortality in patients with diabetes mellitus as much as four times. It has been well established that irisin, with its ability to regulate glucose and lipid homeostasis as well as anti-inflammatory and anti-apoptotic properties, has been widely examined for its therapeutic potentials in managing metabolic disorders. However, the mechanism of irisin in the regulation of cerebral ischemic stroke remains unclear. Using PC12 cells as a model, we have shown that hypoxia/reoxygenation inhibits cell viability and increases lactic dehydrogenase. Irisin, in a dose-dependent manner, reversed these changes. The increase in inflammatory mediators (IL-1β, IL-6, and TNF-α) by hypoxia/reoxygenation was reversed by irisin. Furthermore, the cell apoptosis promoted by hypoxia/reoxygenation was also inhibited by irisin. Irisin suppressed TLR4/MyD88 signaling pathway leading to amelioration of inflammation and apoptosis in PC12 cells. Thus, inhibition of TLR4/MyD88 signaling pathway via irisin could be an important mechanism in the regulation of hypoxia/reoxygenation-induced inflammation and apoptosis in PC12 cells.

Poly-L-arginine: Enhancing Cytotoxicity and Cellular Uptake of Doxorubicin and Necrotic Cell Death

2019 ◽  
Vol 18 (10) ◽  
pp. 1448-1456 ◽  
Author(s):  
Bahareh Movafegh ◽  
Razieh Jalal ◽  
Zobeideh Mohammadi ◽  
Seyyede A. Aldaghi
Keyword(s):  
Cell Death ◽  
Cellular Uptake ◽  
Combined Treatment ◽  
Annexin V ◽  
Cell Penetrating Peptides ◽  
Short Exposure ◽  
Dependent Manner ◽  
Du145 Cells ◽  
Induced Apoptosis ◽  
Resistance To Doxorubicin

Objective: Cell resistance to doxorubicin and its toxicity to healthy tissue reduce its efficiency. The use of cell-penetrating peptides as drug delivery system along with doxorubicin is a strategy to reduce its side effects. In this study, the influence of poly-L-arginine on doxorubicin cytotoxicity, its cellular uptake and doxorubicin-induced apoptosis on human prostate cancer DU145 cells are assessed. Methods: The cytotoxicity of doxorubicin and poly-L-arginine, alone and in combination, in DU145 cells was evaluated at different exposure times using MTT assay. The influence of poly-L-arginine on doxorubicin delivery into cells was evaluated by fluorescence microscopy and ultraviolet spectroscopy. DAPI and ethidium bromide- acridine orange stainings, flow cytometry using annexin V/propidium iodide, western blot analysis with anti-p21 antibody and caspase-3 activity were used to examine the influence of poly-L-arginine on doxorubicininduced cell death. Results: Poly-L-arginine had no cytotoxicity at low concentrations and short exposure times. Poly-L-arginine increased the cytotoxic effect of doxorubicin in DU145 cells in a time-dependent manner. But no significant reduction was found in HFF cell viability. Poly-L-arginine seems to facilitate doxorubicin uptake and increase its intracellular concentration. 24h combined treatment of cells with doxorubicin (0.5 µM) and poly-L-arginine (1 µg ml-1) caused a small increase in doxorubicin-induced apoptosis and significantly elevated necrosis in DU145 cells as compared to each agent alone. Conclusion: Our results indicate that poly-L-arginine at lowest and highest concentrations act as proliferationinducing and antiproliferative agents, respectively. Between these concentrations, poly-L-arginine increases the cellular uptake of doxorubicin and its cytotoxicity through induction of necrosis.

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