Oxidative stress and cell death in the cerebral cortex as a long-term consequence of neonatal hypoglycemia

2016 ◽  
Vol 94 (9) ◽  
pp. 1015-1022 ◽  
Author(s):  
T.R. Anju ◽  
P.R. Akhilraj ◽  
C.S. Paulose
Keyword(s):  
Oxidative Stress ◽  
Gene Expression ◽  
Cell Death ◽  
Cerebral Cortex ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Neonatal Hypoglycemia ◽  
Old Rats

Neonatal hypoglycemia limits glucose supply to cells leading to long-term consequences in brain function. The present study evaluated antioxidant and cell death factors’ alterations in cerebral cortex of 1-month-old rats exposed to neonatal hypoglycemia. Gene expression studies by real-time PCR were carried out using gene-specific TaqMan probes. Fluorescent dyes were used for immunohistochemistry and nuclear staining and imaged by confocal microscope. Total antioxidant level and expression of antioxidant enzymes — superoxide dismutase (SOD) and gluthathione peroxide (GPx) — mRNA was significantly reduced along with high peroxide level in the cerebral cortex of 1-month-old rats exposed to neonatal hypoglycemia. Real-time PCR analysis showed an upregulation of Bax, caspase 3, and caspase 8 gene expression. Confocal imaging with TOPRO-3 staining and immunohistochemistry with caspase 3 antibody indicated cell death activation. The reduced free radical scavenging capability coupled with the expression of key factors involved in cell death pathway points to the possibility of oxidative stress in the cortex of 1-month-old rats exposed to neonatal hypoglycemia. The observed results indicate the effects of neonatal hypoglycemia in determining the antioxidant capability of cerebral cortex in a later stage of life.

scholarly journals Long Term Effects of Neonatal Hypoglycemia on Muscarinic Receptor Function in the Cerebellum

2017 ◽  
Vol 2 (1) ◽  
Keyword(s):  
Gene Expression ◽  
Signaling Pathways ◽  
Muscarinic Receptor ◽  
Receptor Subtype ◽  
Receptor Function ◽  
Long Term Effects ◽  
Neonatal Hypoglycemia ◽  
Neonatal Stress ◽  
Old Rats

Neonatal stress conditions like hypoglycemia cause brain damage by affecting various signaling pathways thereby causing long term effects on brain functions. A proper understanding of the signaling pathways affected by this stress will help to devise better neonatal care. The focus of the current study was to evaluate the effect of neonatal hypoglycemic insult on cerebellar metabotropic cholinergic receptor function in one month old rats. The receptor analysis of cholinergic muscarinic receptors were done by radioreceptor assays and gene expression was analysed using Real Time PCR. Neonatal hypoglycemia significantly reduced (p<0.001) the cerebellar muscarinic receptor density with a down regulation (p<0.001) of muscarinic M3 receptor subtype gene expression in one month old rats. Both muscarinic M1 and M2 receptor subtype expression were not significantly altered. The catabolic enzyme in acetyl choline metabolism- acetylcholine esterase – showed a significant (p<0.001) up regulation with no siginificant change in the anabolic enzyme – choline acetyl transferase, signifying a change in the turnover ratio. Targeting these pathways at different levels can be exploited to devise better treatment for neonatal stress management and also for diseases with impaired insulin secretion such as diabetes.

scholarly journals AN INVESTIGATION INTO THE EFFECT OF H9N2 INFLUENZA VIRUS ON APOPTOSIS OF TESTICULAR CELLS IN CHICKEN EMBRYO USING SYBR-GREEN REAL-TIME PCR

2020 ◽  
Vol 17 (34) ◽  
pp. 1003-1011
Author(s):  
Roshanak Bahrami NAZARABADI ◽  
Mohammad Javad MEHRABANPOUR ◽  
Mohammad Amin EDALATMANESH ◽  
Mehrdad SHARIATI
Keyword(s):  
Gene Expression ◽  
Cell Death ◽  
Influenza Virus ◽  
Real Time ◽  
Real Time Pcr ◽  
Testicular Tissue ◽  
H9n2 Influenza Virus ◽  
Testicular Cells ◽  
Molecular Studies ◽  
Apoptotic Genes

Influenza viruses can induce cell death in their host through apoptosis or necrosis.The H9N2 is a subtype of the Avian Influenza Virus (AIV) that can cause severe damage to reproductive organs of laying hens. The present study aimed to investigate the effect of the H9N2 influenza virus on apoptosis of testicular cells in chicken embryos. To this end, A/Chicken/Tehran/ZMT-101/99(H9N2) was inoculated to 210 embryonated fifteen-day-old eggs laid by SPF hens. Then, according to the experiment design, live embryos were dissected on the 19th and 21st days of fetal life and on the 25th day after birth for pathological and molecular studies and evaluation of gene expression in testicular tissue. Dissected tissues were stained with hematoxylin and eosin (HE) and studied under an optical microscope at 400x magnification. For molecular studies, viral RNA was extracted from testicular tissues, replicated by RT-PCR, and finally evaluated H9N2. Then, the genes expression involved in the testicular tissue cells apoptosis was evaluated through real-time PCR. Pathological studies indicated that H9N2 caused lesions in testicular tissue orchitis, seminiferous, and nephritis of the host. Molecular studies also showed that H9N2 replication in the host body increases BAX and Caspase 3 expression and reduces the expression of Bcl-2 and Bcl-XL. These changes in gene expression increased apoptosis and induced cell death in the host. In summary, the study findings suggested that H9N2 can increase the expression of pro-apoptotic genes and reduce the expression of anti-apoptotic genes, resulting in severe destruction of testicular tissue caused by cell apoptosis.

Effect of Mir-663 on the Differentiation of Bone Marrow Mesenchymal Stem Cells in Inflammatory Environment Through Regulating Oxidative Stress and Transforming Growth Factor β1

2019 ◽  
Vol 9 (11) ◽  
pp. 1600-1606
Author(s):  
Xiuhong Sun ◽  
Yujie Liu ◽  
Shujuan Zheng
Keyword(s):  
Oxidative Stress ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β1 ◽  
Control Group ◽  
Sod Activity ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

BMSCs are beneficial for the treatment of Osteoarthritis (OA). Mir-663 involves in various diseases. However, the role of Mir-663 in the differentiation of BMSCs in an inflammatory environment remains unclear. Rat BMSCs were isolated and divided into control group, and inflammation group which was treated with 1 μg/ml lipopolysaccharide (LPS), Mir-663 group and Mir-663 siRNA groups which was respectively transfected with Mir-663 plasmid and Mir-663 siRNA into LPS-treated BMSCs followed by analysis of the survival rate of BMSC by MTT, Caspase 3 activity, ALP activity, expression of osteogenic genes Runx2 and OP by Real time PCR, ROS content and SOD activity, TGF-β1 level by Real time PCR and ELISA, TNF- and IL-1 secretion by ELISA. LPS treatment significantly increased Mir-663 expression, inhibited cell proliferation, increased Caspase 3 activity, and inhibited ALP activity. Meanwhile it also significantly decreased Runx2 and OP expression, increased ROS content, decreased SOD activity and TGF-β1 expression as well as elevated TNF-α and IL-1β secretion compared to control (P < 0.05); Mir-663 plasmid transfection can further promote the above changes (P < 0.05); whereas, Mir-663 siRNA could reverse the above changes (P < 0.05). Mir-663 expression was increased in BMSCs in inflammatory environment. Down-regulation of Mir663 expression can regulate oxidative stress, up-regulate TGF-β1 level, inhibit the secretion of TNF-α and IL-1β, and promote the proliferation and osteogenic differentiation of BMSCs in inflammatory environment.

Cortical cholinergic dysregulation as a long-term consequence of neonatal hypoglycemia

2015 ◽  
Vol 93 (1) ◽  
pp. 47-53 ◽  
Author(s):  
T.R. Anju ◽  
C.S. Paulose
Keyword(s):  
Cerebral Cortex ◽  
Life Stress ◽  
Energy Demand ◽  
High Energy ◽  
Acetylcholine Esterase ◽  
Long Term Effects ◽  
Neonatal Hypoglycemia ◽  
Acetylcholine Metabolism ◽  
Old Rats

Neonatal hypoglycemia limits the glucose supply to cells, affecting the function of brain due to its high energy demand. This can cause long-term consequences in brain function, leading to memory and cognitive deficits. The present study evaluated the cholinergic functional regulation in cerebral cortex of one month old rats exposed to neonatal hypoglycemia to understand the long-term effects of early life stress. Receptor binding and gene expression studies were done in the cerebral cortex to analyze the changes in total muscarinicreceptors, muscarinic M1, M2, M3 receptors, and the enzymes involved in acetylcholine metabolism, cholineacetyl transferase and acetylcholine esterase. Neonatal hypoglycemia decreased total muscarinic receptors (p < 0.001) with reduced muscarinic M1, M2, and M3 receptor genes (p < 0.001) in one month old rats. The reduction in acetylcholine metabolism is indicated by the downregulated cholineacetyl transferase, upregulated acetylcholine esterase, and decreased vesicular acetylcholine transporter expression. These alterations in cholinergic function in one month old rat brain indicates the longterm consequences of neonatal hypoglycemia in cortical function, which can contribute to the onset of many disease conditions in later stages of life.

Identification of adeno-associated viral vectors suitable for intestinal gene delivery and modulation of experimental colitis

2012 ◽  
Vol 302 (3) ◽  
pp. G296-G308 ◽  
Author(s):  
Steven Polyak ◽  
Annette Mach ◽  
Stacy Porvasnik ◽  
Lisa Dixon ◽  
Thomas Conlon ◽  
...  
Keyword(s):  
Gene Expression ◽  
Small Intestine ◽  
Gene Transfer ◽  
Real Time ◽  
Real Time Pcr ◽  
Transgene Expression ◽  
Intestinal Inflammation ◽  
Experimental Colitis ◽  
Mucosal Inflammation

Effective gene transfer with sustained gene expression is an important adjunct to the study of intestinal inflammation and future therapy in inflammatory bowel disease. Recombinant adeno-associated virus (AAV) vectors are ideal for gene transfer and long-term transgene expression. The purpose of our study was to identify optimal AAV pseudotypes for transduction of the epithelium in the small intestine and colon, which could be used for studies in experimental colitis. The tropism and transduction efficiencies of AAV pseudotypes 1–10 were examined in murine small intestine and colon 8 wk after administration by real-time PCR and immunohistochemistry. The clinical and histopathological effects of IL-10-mediated intestinal transduction delivered by AAVrh10 were examined in the murine IL-10−/− enterocolitis model. Serum IL-10 levels and IL-10 expression were followed by ELISA and real-time PCR, respectively. AAV pseudotypes 4, 7, 8, 9, and 10 demonstrated optimal intestinal transduction. Transgene expression was sustained 8 wk after administration and was frequently observed in enteroendocrine cells. Long-term IL-10 gene expression and serum IL-10 levels were observed following AAV transduction in an IL-10−/− model of enterocolitis. Animals treated with AAVrh10-IL-10 had lower disease activity index scores, higher colon weight-to-length ratios, and lower microscopic inflammation scores. This study identifies novel AAV pseudotypes with small intestine and colon tropism and sustained transgene expression capable of modulating mucosal inflammation in a murine model of enterocolitis.

Respon Morfologis dan Ekspresi Gen Aquaporin pada Padi IR 64 yang Mengalami Cekaman Kekeringan pada Fase Reproduktif (Morphological Responses and Aquaporin Gene Expression in Rice IR64 under Drought Stress at the Reproductive Stage)

2018 ◽  
Vol 7 (2) ◽  
Author(s):  
Made Pharmawati ◽  
Ni Nyoman Wirasiti ◽  
Luh Putu Wrasiati
Keyword(s):  
Gene Expression ◽  
Drought Stress ◽  
Real Time ◽  
Real Time Pcr ◽  
Control Plant ◽  
Reproductive Stage ◽  
Limiting Factors ◽  
Rice Grain ◽  
Aquaporin Gene ◽  
Peg Treatment

Abstrak Cekaman kekeringan merupakan faktor pembatas penting bagi pertumbuhan dan produktivitas tanaman termasuk padi.      Penelitian ini bertujuan menganalisis respon padi IR64 terhadap cekaman kekeringan dengan pemberian polietilen glikol (PEG) pada fase reproduktif.  Penelitian juga bertujuan menganalisis ekspresi gen aquaporin akibat cekaman kekeringan.  Bibit padi ditanam dalam pot dan perlakuan PEG dengan konsentrasi 108g/L (-0.25MPa) dan 178g/L (-0.52 MPa) diberikan saat munculnya panikula. Perlakuan diberikan selama 2 minggu, kemudian tanaman disiram kembali.  Ekspresi gen diamati pada akhir perlakuan dengan semi kuantitatif real time PCR.  Ekstraksi RNA menggunakan RNeasy plant mini kit, sedangkan sintesis cDNA menggunakan Transcriptor First Strand cDNA Kit.  Hasil penelitian menunjukkan bahwa jumlah malai dan berat total malai berkurang akibat cekaman kekeringan.  Persentase gabah kosong mencapai 84,6% pada perlakuan PEG-0,52 MPa, sedangkan pada perlakuan PEG -0,25 MPa persentase gabah kosong sebesar 67,8%.  Pada kontrol persentase gabah kosong adalah 10,3%.  Ekspresi gen OsPIP2;7 sedikit menurun pada perlakuan PEG -0,52 MPa.Kata kunci: ekspresi gen, IR64, kekeringan, padi, PEG  Abstract Drought stress is one of the limiting factors of plant growth and productivity including rice.  The aim of this study was to analyze responses of IR64 rice to polyethylene glycol (PEG)-induced-drought stress at the reproductive stage.  This study also aimed to analyze the expression of aquaporin under drought stress.  Rice seedlings were grown in pot system and PEG treatment at concentration of -0.25MPa (108g/L) and -0.52 MPa (178g/L) were given when the panicles arose.  Treatments were conducted for 2 weeks, after that the plants were rewatered.  Gene expression was evaluated at the end of PEG treatment using semi quantitative real time PCR. RNA was extracted using RNeasy plant mini kit, while cDNA synthesis was done using Transcriptor First Strand cDNA Kit.  The results showed that the number and weight of rice ear were less in plant treated with PEG than in control.  The percentage of empty rice grain reached 84.6% at PEG -0.52 MPa, while at PEG -0.25 MPa the percentage of empty grain was 67.8%.  In control plant, the percentage of empty grain was 10.3%.  Drought stress did not alter the expression of OsPIP2;7.  Keywords: drought, gene expression, IR64, PEG, rice

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