Exogenous spermidine affects polyamine metabolism in the mouse hypothalamus

Abstract Spermidine is important for the hypothalamic control of pituitary secretion of hormones involved in neuroendocrine functions in mammals. In this study, the effect of exogenous spermidine on the expression of genes and proteins related to polyamine metabolism and polyamine levels was examined. The results indicated that treatment with spermidine at 0.05 mg/g (BW) significantly increased the levels of Oaz1 mRNA and protein expression and decreased putrescine content in mouse hypothalamus (p < 0.05). The administration with spermidine at 0.10 mg/g significantly increased the levels of Oaz1, Oaz2, and Odc expression in mouse hypothalamus (p < 0.05). Treatment with spermidine at 0.05 mg/g significantly increased the levels of Ssat mRNA expression and reduced the level of Smo mRNA expression in mouse hypothalamus (p < 0.05). Putrescine concentrations in the hypothalamus after the administration of spermidine at 0.10 and 0.15 mg/g were significantly higher than those in the control group (p < 0.05). The concentration of both spermidine and spermine in the hypothalamus after the administration of spermidine at 0.15 mg/g was decreased significantly (p < 0.05). In summary, our results indicate that exogenous spermidine affects polyamine homeostasis in the mouse hypothalamus by modulating the expression of genes and proteins related to polyamine metabolism.

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Altered mRNA and Protein Expression of Monocarboxylate Transporter MCT1 in the Cerebral Cortex and Cerebellum of Prion Protein Knockout Mice

International Journal of Molecular Sciences ◽

10.3390/ijms22041566 ◽

2021 ◽

Vol 22 (4) ◽

pp. 1566

Author(s):

Sanja Ramljak ◽

Matthias Schmitz ◽

Cendrine Repond ◽

Inga Zerr ◽

Luc Pellerin

Keyword(s):

Protein Expression ◽

Mrna Expression ◽

Prion Protein ◽

Mrna Level ◽

Brain Regions ◽

Cellular Prion Protein ◽

Monocarboxylate Transporter ◽

Control Group ◽

Expression Of Genes ◽

Subunit Expression

The effect of a cellular prion protein (PrPc) deficiency on neuroenergetics was primarily analyzed via surveying the expression of genes specifically involved in lactate/pyruvate metabolism, such as monocarboxylate transporters (MCT1, MCT2, MCT4). The aim of the present study was to elucidate a potential involvement of PrPc in the regulation of energy metabolism in different brain regions. By using quantitative real-time polymerase chain reaction (qRT-PCR), we observed a marked reduction in MCT1 mRNA expression in the cortex of symptomatic Zürich I Prnp−/− mice, as compared to their wild-type (WT) counterparts. MCT1 downregulation in the cortex was accompanied with significantly decreased expression of the MCT1 functional interplayer, the Na+/K+ ATPase α2 subunit. Conversely, the MCT1 mRNA level was significantly raised in the cerebellum of Prnp−/− vs. WT control group, without a substantial change in the Na+/K+ ATPase α2 subunit expression. To validate the observed mRNA findings, we confirmed the observed change in MCT1 mRNA expression level in the cortex at the protein level. MCT4, highly expressed in tissues that rely on glycolysis as an energy source, exhibited a significant reduction in the hippocampus of Prnp−/− vs. WT mice. The present study demonstrates that a lack of PrPc leads to altered MCT1 and MCT4 mRNA/protein expression in different brain regions of Prnp−/− vs. WT mice. Our findings provide evidence that PrPc might affect the monocarboxylate intercellular transport, which needs to be confirmed in further studies.

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The Acute Effects of Swimming Exercise on PGC-1α-FNDC5/Irisin-UCP1 Expression in Male C57BL/6J Mice

Metabolites ◽

10.3390/metabo11020111 ◽

2021 ◽

Vol 11 (2) ◽

pp. 111

Author(s):

Eunhee Cho ◽

Da Yeon Jeong ◽

Jae Geun Kim ◽

Sewon Lee

Keyword(s):

Adipose Tissue ◽

Protein Expression ◽

Swimming Exercise ◽

Enzyme Linked Immunosorbent Assay ◽

Control Group ◽

Brown Adipose ◽

Mrna And Protein Expression ◽

Ucp1 Expression ◽

Exercise Induced ◽

Gastrocnemius Muscles

Irisin is a myokine primarily secreted by skeletal muscles and is known as an exercise-induced hormone. The purpose of this study was to determine whether the PGC-1α -FNDC5 /Irisin-UCP1 expression which is an irisin-related signaling pathway, is activated by an acute swimming exercise. Fourteen to sixteen weeks old male C57BL/6J mice (n = 20) were divided into control (CON, n = 10) and swimming exercise groups (SEG, n = 10). The SEG mice performed 90 min of acute swimming exercise, while control (non-exercised) mice were exposed to shallow water (2 cm of depth) for 90 min. The mRNA and protein expression of PGC-1α, FNDC5 and browning markers including UCP1 were evaluated by quantitative real-time PCR and western blotting. Serum irisin concentration was measured by enzyme-linked immunosorbent assay. An acute swimming exercise did not lead to alterations in the mRNA and protein expression of PGC-1α in both soleus and gastrocnemius muscles, the mRNA and protein expression of UCP1 in brown adipose tissue, mRNA browning markers in visceral adipose tissue and circulating irisin when compared with the control group. On the other hand, an acute swimming exercise led to increases in the mRNA and protein expressions of FNDC5 in the soleus muscle, the protein expression of FNDC5 in the gastrocnemius muscles and the protein expression of UCP1 in subcutaneous adipose tissue.

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The effect of miR-6523a on growth hormone secretion in pituitary cells of Yanbian yellow cattle

Canadian Journal of Animal Science ◽

10.1139/cjas-2019-0168 ◽

2020 ◽

Vol 100 (4) ◽

pp. 657-664

Author(s):

Jiuxiu Ji ◽

Taihua Jin ◽

Rui Zhang ◽

Angang Lou ◽

Yingying Chen ◽

...

Keyword(s):

Growth Hormone ◽

Protein Expression ◽

Luciferase Reporter ◽

Control Group ◽

Pituitary Cells ◽

Expression Levels ◽

Gh Secretion ◽

Yellow Cattle ◽

Mrna And Protein Expression ◽

In Cells

Yanbian yellow cattle breeding is limited by its slow growth. We previously found that the miRNA miR-6523a is differentially expressed between Yanbian yellow cattle and Han Yan cattle, which differ in growth characteristics. In this study, we evaluated the effects of miR-6523a on growth hormone (GH) secretion in pituitary cells of Yanbian yellow cattle. Bioinformatics analyses using TargetScan and RNAhybrid, as well as dual luciferase reporter assays, showed that miR-6523a targets the 3′ untranslated region of somatostatin receptor 5 (SSTR5). We further found that the mRNA and protein expression levels of GH in pituitary cells were significantly higher in cells treated with miR-6523a mimic than in the control group (P = 0.0082 and P = 0.0069). The GH mRNA and protein expression levels were lower in cells treated with miR-6523a inhibitor than in the control group, but the difference was not significant (P = 0.064 and P = 0.089). SSTR5 mRNA and protein levels were inhibited by miR-6523a mimic compared with the control group (P = 0.0024 and P = 0.0028) and were elevated slightly by miR-6523a inhibitor (P = 0.093 and P = 0.091). These results prove that miR-6523a regulates GH secretion in pituitary cells by SSTR5. More broadly, these findings provide a basis for studies of the roles of miRNAs in animal growth and development.

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NH4Cl affects the expression of Wnt/β-catenin pathway in hepatocytes

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2017-0604 ◽

2018 ◽

Vol 96 (3) ◽

pp. 281-286

Author(s):

Lu Bai ◽

Jingjing Li ◽

Xiaorui Liu ◽

Shasha Li ◽

Fulei Li ◽

...

Keyword(s):

Protein Expression ◽

Control Group ◽

Cyclin D ◽

Chang Liver Cell ◽

Expression Levels ◽

Low Concentrations ◽

Short Period ◽

Mrna And Protein Expression ◽

High Concentrations ◽

Chang Liver

We intended to explore whether NH4Cl influences the viability and regulates the expression of Wnt/β-catenin pathway in hepatocytes. The Chang liver cell line was used and cultured with different concentrations of NH4Cl (2.5, 5, 10, 20, 40, and 50 mmol/L) for 12, 24, and 48 h. The viability of hepatocytes was detected by MTT assay. The mRNA and protein expression level was analyzed with qRT–PCR and Western blotting, respectively. NH4Cl concentration significantly affects the viability of hepatocytes. With the increase of NH4Cl concentration, the viability of hepatocytes was decreased, accordingly. The mRNA and protein expression of Wnt1, β-catenin, and cyclin D was significantly increased after treatment with low concentrations of NH4Cl as compared with the control group, whereas their expression levels were decreased after treatment with high concentrations of NH4Cl. The mRNA and protein expression of Wnt1, β-catenin, and cyclin D was also significantly increased after treatment with NH4Cl for a short period as compared with the control group, whereas their expression levels were decreased after treatment with NH4Cl for a long period. In addition, we found NH4Cl treatment significantly reversed the results after RNA silencing of Wnt1 in hepatocytes. NH4Cl influences the viability of hepatocytes and affects the expression of Wnt/β-catenin pathway in hepatocytes.

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Differential between Protein and mRNA Expression of CCR7 and SSTR5 Receptors in Crohn's Disease Patients

Mediators of Inflammation ◽

10.1155/2009/285812 ◽

2009 ◽

Vol 2009 ◽

pp. 1-9 ◽

Cited By ~ 10

Author(s):

Nathalie Taquet ◽

Serge Dumont ◽

Jean-Luc Vonesch ◽

Didier Hentsch ◽

Jean-Marie Reimund ◽

...

Keyword(s):

Crohn’S Disease ◽

Crohn's Disease ◽

Protein Expression ◽

Mrna Expression ◽

Large Scale ◽

Mononuclear Cells ◽

Biologically Active ◽

Chronic Inflammatory Bowel Disease ◽

Peripheral Blood Mononuclear ◽

Mrna And Protein Expression

Crohn's disease (CD) is a multifactorial chronic inflammatory bowel disease of unknown cause. The aim of the present study was to explore if mRNA over-expression of SSTR5 and CCR7 found in CD patients could be correlated to respective protein expression. When compared to healthy donors, SSTR5 was over-expressed 417±71 times in CD peripheral blood mononuclear cells (PBMCs). Flow cytometry experiments showed no correlation between mRNA and protein expression for SSTR5 in PBMCs. In an attempt to find a reason of such a high mRNA expression, SSTR5 present on CD PBMCs were tested and found as biologically active as on healthy cells. In biopsies of CD intestinal tissue, SSTR5 was not over-expressed but CCR7, unchanged in PBMCs, was over-expressed by 10±3 times in the lamina propria. Confocal microscopy showed a good correlation of CCR7 mRNA and protein expression in CD intestinal biopsies. Our data emphasize flow and image cytometry as impossible to circumvent in complement to molecular biology so to avoid false interpretation on receptor expressions. Once confirmed by further large-scale studies, our preliminary results suggest a role for SSTR5 and CCR7 in CD pathogenesis.

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Parathyroid hormone stimulates endothelial expression of atherosclerotic parameters through protein kinase pathways

AJP Renal Physiology ◽

10.1152/ajprenal.00406.2006 ◽

2007 ◽

Vol 292 (4) ◽

pp. F1215-F1218 ◽

Cited By ~ 77

Author(s):

Gloria Rashid ◽

Jacques Bernheim ◽

Janice Green ◽

Sydney Benchetrit

Keyword(s):

Parathyroid Hormone ◽

Protein Kinase ◽

Protein Expression ◽

Mrna Expression ◽

Umbilical Vein ◽

Protein Levels ◽

Glycation End Products ◽

Mrna And Protein Expression ◽

Vascular Structures ◽

The Impact

Parathyroid hormone (PTH), the major systemic calcium-regulating hormone, has been linked to uremic vascular changes. Considering the possible deleterious action of PTH on vascular structures, it seemed logical to evaluate the impact of PTH on the receptor of advanced glycation end products (RAGE) and interleukin 6 (IL-6) mRNA and protein expression, taking into account that such parameters might be involved in the pathogenesis of vascular calcification, atherosclerosis, and/or arteriolosclerosis. Human umbilical vein cord endothelial cells (HUVEC) were stimulated for 24 h with 10−12–10−10 mol/l PTH. The mRNA expression of RAGE and IL-6 was established by reverse transcriptase/PCR techniques. RAGE protein levels were determined by Western blot and IL-6 secretion was measured by ELISA. The pathways by which PTH may have an effect on HUVEC functions were evaluated. PTH (10−11–10−10mol/l) significantly increased RAGE mRNA and protein expression. PTH also significantly increased IL-6 mRNA expression without changes at protein levels. The addition of protein kinase (PKC or PKA) inhibitors or nitric oxide (NO) synthase inhibitors significantly reduced the RAGE and IL-6 mRNA expression and the RAGE protein expression. PTH stimulates the mRNA expressions of RAGE and IL-6 and the protein expression of RAGE. These stimulatory effects are probably through PKC and PKA pathways and are also NO dependent. Such data may explain the possible impact of PTH on the atherosclerotic and arteriosclerotic progression.

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Increased galectin-3 levels are associated with abdominal aortic aneurysm progression and inhibition of galectin-3 decreases elastase-induced AAA development

Clinical Science ◽

10.1042/cs20171142 ◽

2017 ◽

Vol 131 (22) ◽

pp. 2707-2719 ◽

Cited By ~ 13

Author(s):

Carlos-Ernesto Fernandez-García ◽

Carlos Tarin ◽

Raquel Roldan-Montero ◽

Diego Martinez-Lopez ◽

Monica Torres-Fonseca ◽

...

Keyword(s):

Abdominal Aortic Aneurysm ◽

Aortic Aneurysm ◽

Protein Expression ◽

Potential Role ◽

Control Group ◽

Mrna Levels ◽

Abdominal Aortic ◽

Stat3 Phosphorylation ◽

Galectin 3 ◽

Mrna And Protein Expression

Abdominal aortic aneurysm (AAA) evolution is unpredictable and no specific treatment exists for AAA, except surgery to prevent aortic rupture. Galectin-3 has been previously associated with CVD, but its potential role in AAA has not been addressed. Galectin-3 levels were increased in the plasma of AAA patients (n=225) compared with the control group (n=100). In addition, galectin-3 concentrations were associated with the need for surgical repair, independently of potential confounding factors. Galectin-3 mRNA and protein expression were increased in human AAA samples compared with healthy aortas. Experimental AAA in mice was induced via aortic elastase perfusion. Mice were treated intravenously with the galectin-3 inhibitor modified citrus pectin (MCP, 10 mg/kg, every other day) or saline. Similar to humans, galectin-3 serum and aortic mRNA levels were also increased in elastase-induced AAA mice compared with control mice. Mice treated with MCP showed decreased aortic dilation, as well as elastin degradation, vascular smooth muscle cell (VSMC) loss, and macrophage content at day 14 postelastase perfusion compared with control mice. The underlying mechanism(s) of the protective effect of MCP was associated with a decrease in galectin-3 and cytokine (mainly CCL5) mRNA and protein expression. Interestingly, galectin-3 induced CCL5 expression by a mechanism involving STAT3 activation in VSMC. Accordingly, MCP treatment decreased STAT3 phosphorylation in elastase-induced AAA. In conclusion, increased galectin-3 levels are associated with AAA progression, while galectin-3 inhibition decreased experimental AAA development. Our data suggest the potential role of galectin-3 as a therapeutic target in AAA.

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Expression of Pulmonary Surfactant-Associated Protein B in Neonatal Respiratory Distress Syndrome

Journal of Medical Biochemistry ◽

10.2478/jomb-2013-0005 ◽

2013 ◽

Vol 32 (2) ◽

pp. 146-151

Author(s):

Xiaojuan Yin ◽

Yan Wang ◽

Lu Xie ◽

Xiangyong Kong ◽

Chunzhi Wang ◽

...

Keyword(s):

Protein Expression ◽

Respiratory Distress Syndrome ◽

Respiratory Distress ◽

Mrna Expression ◽

Pulmonary Surfactant ◽

Distress Syndrome ◽

Neonatal Respiratory Distress Syndrome ◽

Control Group ◽

Neonatal Respiratory Distress ◽

Protein B

Summary Background: The aim of this study was to investigate the role of pulmonary surfactant-associated protein B (SP-B) expression in the pathogenesis of neonatal respiratory distress syndrome (RDS) via detecting the protein and mRNA expression of SP-B. Methods: A total of 60 unrelated neonates who died of RDS were chosen as the RDS group and then subgrouped into ≤32 weeks group, 32∼37 weeks group and ≥37 weeks group (n=20). Sixty neonates who died of other diseases were enrolled as controls and subdivided into 3 matched groups based on the gestational age. Western blot assay and RT-PCR were employed. Results: In the RDS group, SP-B protein expression was reduced or deficient in 8 neonates of which 6 had no SP-B protein expression. In the control group, only 1 had reduced SP-B protein expression. The reduced or deficient SP-B protein expression in 9 neonates of both groups was noted in the ≥37 weeks group. In the RDS group, the SP-B mRNA expression was significantly lower than that in the control group. In the ≤37 weeks group, SP-B mRNA expression was comparable between the RDS group and control group. In the 32∼37 weeks group, the SP-B mRNA expression in the RDS group was significantly reduced when compared with the control group. In the ≥37 weeks group, the SP-B mRNA expression in the RDS group was dramatically lower than that in the control group. Conclusions: Alteration of SP-B expression is present at transcriptional and translational levels. Reduction of SP-B mRNA and protein expression is involved in the pathogenesis of RDS.

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SUZ12 Participates in PNH Cloning Proliferation By Regulating Histone H3K27me3 Methylation Level

Blood ◽

10.1182/blood-2021-148546 ◽

2021 ◽

Vol 138 (Supplement 1) ◽

pp. 1106-1106

Author(s):

Rong Fu ◽

Yingying Chen ◽

Zonghong Shao ◽

Hui Liu ◽

Lijie Zeng ◽

...

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Protein Expression ◽

Peripheral Blood ◽

Methylation Level ◽

Cell Model ◽

Gene Mutations ◽

Control Group ◽

Mrna And Protein Expression ◽

And Control

Abstract Paroxysmal nocturnal hemoglobinuria (PNH) is a disease of hematopoietic stem cell membrane defects due to acquired PIG-Amutation. Our previous study found some secondary gene mutations in PNH patients by WES. However, it is not clear exactly which mutations are associated with the disease. So, 97 target genes were selected as a target gene panel and tested in 23 PNH patients by DNA sequencing of specific target regions. We found that all PNH patients had other gene mutations except PIG-Amutations, including TTN, NCOR2, CPS1, MUC4, SUZ12, LFNG, CELSR2, JAK2, SETBP1 and KMT2D (Figure1A). Through harmful analysis, KEGG enrichment, GO enrichment analysis and protein interaction analysis, we screened out the secondary mutant gene SUZ12 that may be involved in the cloning proliferation of PNH. We detected the mRNA and protein expression levels of SUZ12 and H3K27me3 methylation in PNH patients and health volunteers, the results showed that the mRNA and protein expression levels of SUZ12 and H3K27me3 methylation in peripheral blood CD59 -cells of PNH patients were higher than those in CD59 + cells of PNH patients and healthy controls (Figure1B). The relative expression level of SUZ12 in peripheral blood CD59 -cells of PNH patients was correlated with (r=0.4162, p=0.0385), CD59 -erythrocyte ratio (r=0.4636, p=0.0196), CD59 -monocyte ratio (r=0.4052, p=0.0495), Flaer -monocyte ratio (r=0.6769, p=0.0004) and Flaer -granulocytic ratio (r=0.6146, p=0.0018), indicating that SUZ12 may be involved in abnormal PNH cloning and proliferation by regulating H3K27me3. To verify the role of SUZ12 in the proliferation of PNH cloning, we used CRISPR/Cas9 to knockdown PIG-A expression in THP-1 cells to construct A PNH cell model, the expression level of PIG-A protein in the cell model was significantly decreased, and the proportion of CD59 - cells accounted was stable at 95%. Then lentivirus transfection was used to knockdown the expression of SUZ12 in PNH cell model. The results showed when the SUZ12 expression was knockdown, the methylation level of histone H3K27me3 was decreased, the cell proliferation activity was decreased, apoptosis was increased, and the cell cycle was arrested at G0/G1 phase. The proportion of CD59 + cells increased gradually from 3 weeks after transfection, and significantly increased at 4 weeks after transfection, while no changes were observed in the empty virus group and control group (Figure1C). Four weeks after lentivirus transfection, the expression of PIG-A protein recovered in SUZ12 knockdown group compared with empty virus group and control group (Figure1D). In conclusion, SUZ12 mutation leads to the overexpression of SUZ12, which can affect cell proliferation, apoptosis and cell cycle by regulating the methylation level of histone H3K27me3, thereby promoting the proliferation of PNH abnormal cloning and participating in the pathogenesis of PNH. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

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PO-298 Effect of exercise and Siyeshen upon hippocampal cytochrome c and caspase-3 of diabetic rats

Exercise Biochemistry Review ◽

10.14428/ebr.v1i5.13453 ◽

2018 ◽

Vol 1 (5) ◽

Author(s):

Dongmei Wang ◽

Jinming Zhang ◽

Haibin Liu ◽

Rongmei Wang

Keyword(s):

Cytochrome C ◽

Protein Expression ◽

Caspase 3 ◽

Diabetic Control ◽

Exercise Group ◽

Diabetic Rats ◽

Control Group ◽

Diabetic Control Group ◽

Mrna And Protein Expression ◽

Cyt C

Objective To observe the effects of aerobic exercise and Siyeshen water extract on cytochrome c (Cyt c) and caspase-3 in hippocampus of diabetic rats and to explore the possible mechanism of improving diabetes. Methods Healthy male Wister rats fed with high fat and high sugar and combined with streptozotocin to establish type II diabetes model. They were randomly divided into 4 groups: diabetic control group, exercise group, Siyeshen group and exercise+Siyeshen group, and another normal control group, with 6 rats in each group. After aerobic exercise (15m/min, 5°slope, 60min, every other day) or/and Siyeshen (200mg/kg) gastrointestinal administration for 8w, the expression of Cyt c and caspase-3 in hippocampus of each group were detected by immunoblotting, and mRNA expressions were detected by RT-PCR. Results Compared with the normal control group, the mRNA and protein expressions of Cyt c and caspase-3 in the hippocampus of the diabetic control group were significantly increased (P<0.05). Compared with the diabetic control group, the blood glucose level of exercise group and exercise+ Siyeshen group decreased (P<0.05), the mRNA and protein expression of Cyt c and caspase-3 decreased significantly (P<0.05), but there were no significant changes in the mRNA and protein expression of Cyt c and caspase-3 between Siyeshen group and diabetic control group (P﹥0.05). Conclusions Exercise and exercise combined with Siyeshen can inhibit cytochrome c release and reduce caspase-3 protein expression, which may be related to the improvement of blood glucose levels in diabetic rats.

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