scholarly journals CORM-2 can Attenuate Bleeding-mediated Inflammation by Increasing Phagocytic Capacity of Cerebral Microglial Cells in Neonatal Rat in Vitro

2021 ◽  
Author(s):  
Zhiying Chen ◽  
Huiyan Zhang ◽  
Jun Zhou ◽  
Xiaoqin Wu ◽  
Moxin Wu ◽  
...  
Keyword(s):  
Treated Group ◽  
Neonatal Rat ◽  
Neonatal Rats ◽  
Phagocytic Capacity ◽  
Inflammatory Reactions ◽  
Microglial Phagocytosis ◽  
Protein Expressions ◽  
Cell Groups ◽  
Cell Supernatant

Abstract Objective: This study aimed to explore the mechanism of CORM-2 on attenuating bleeding-related inflammation. Methods: Microglia were isolated from the neonatal rats (1-2days old) and identified by the CD11b/c anti-body, and some microglia were co-cultured with RBCs marked with PKH26 fluorescent dye, and then treated with CORM-2. That is, the microglia cells were divided into the microglia, microglia+ PKH26+RBCs and microglia + PKH26+ RBCs+CORM-2 cell-groups. Microglial phagocytosis to RBCs PKH26+ was observed under an inverted fluorescence microscope; moreover, the fluorescence intensity of microglia that phagocytized PKH26+RBCs was detected through immunofluorescence. HO-1, NF-κB p65, and IL-1β expressions were detected using RT-qPCR, western blotting, and immunofluorescence, respectively. The levels of carbon monoxide hemoglobin (HbCO) in the cell supernatant in each group were detected with ELISA.Results. After 1- day of co-culturing, the number of residual PKH26+RBCs in the Microglia+ PKH26+RBCs+CORM-2 group decreased remarkably than that in the Microglia+ PKH26+RBCs groups (18 × 106 vs. 14 × 106, p=0.02), which revealed that microglia phagocytosis was stronger in CORM-2 treated group. More over, compared with microglia + PKH26+RBCs group, the microglia+ PKH26+RBCs +CORM-2 group showed higher levels of HO-1 mRNA and protein expressions at the 3rd day and the 5th day after co-culturing. Further more, CORM-2 significantly inhibited the expressions of mRNA and proteins of NF‐κB p65 and IL-1 after 3 days of co-culturing, meanwhile, CORM-2 did not increase the level of HbCO in the cell supernatant.Conclusions CORM-2 can inhibit inflammatory reactions in bleeding setting in vitro by promoting microglial phagocytosis to RBCs and decrease IL-1β and NF-κB; the mechanism may involve HO-1/CO system.

scholarly journals Developmental Expression of the Peripheral-Type Benzodiazepine Receptor and the Advent of Steroidogenesis in Rat Adrenal Glands*

Endocrinology ◽  
1999 ◽  
Vol 140 (2) ◽  
pp. 859-864 ◽  
Author(s):  
Alexandra Zilz ◽  
Hua Li ◽  
Rosa Castello ◽  
Vassilios Papadopoulos ◽  
Eric P. Widmaier
Keyword(s):  
Ligand Binding ◽  
Binding Capacity ◽  
Benzodiazepine Receptor ◽  
Neonatal Rat ◽  
Developmental Expression ◽  
Neonatal Rats ◽  
Adrenal Cells ◽  
Star Protein ◽  
Peripheral Type

Abstract Although the precise mechanism whereby cholesterol is transported across the outer mitochondrial membrane is uncertain, a multimeric receptor complex termed the peripheral-type benzodiazepine receptor (PBR) appears essential for this process. We therefore predicted that adrenal cells at different developmental stages would express PBR coincidentally with the advent of steroidogenesis. Adrenals of neonatal rats demonstrate greatly reduced sensitivity to ACTH that gradually increases after the first 2 weeks of life. Thus, neonates have lower circulating corticosterone levels following exposure to stress. We examined mitochondrial PBR ligand binding activity, immunoreactive (ir) PBR content, and adrenal sensitivity to ACTH in vivo and in vitro. Ontogeny of both mitochondrial PBR ligand binding capacity and irPBR directly paralleled that of ACTH-inducible steroidogenesis in isolated rat adrenal cells and in rats injected with ACTH. In addition, neonatal PBR had approximately 2-fold higher affinity for PK11195, a synthetic ligand that binds with high affinity to PBR. No correlation was observed during neonatal life between ir-steroidogenic acute regulatory (StAR) protein content and steroidogenesis. These results are consistent with the hypothesis that PBR is an absolute prerequisite for adrenocortical steroidogenesis, and suggest that the stress hyporesponsive period of neonatal rats may result from decreased PBR expression. In addition, the higher affinity of neonatal PBR and the relatively high basal expression of StAR protein in neonatal adrenals may partly explain the high constitutive steroidogenesis characteristic of neonatal rat adrenal cells.

scholarly journals Chlorobenzoxime inhibits respiratory syncytial virus infection in neonatal rats via up-regulation of IFN-γ in dendritic cells

2020 ◽  
Vol 19 (2) ◽  
pp. 239-246
Author(s):  
Junzhao Li ◽  
Yonghai Zhang ◽  
Hongmei Qiao ◽  
Yingji Jin ◽  
Jianmin Wang ◽  
...  
Keyword(s):  
Dendritic Cells ◽  
Respiratory Syncytial Virus ◽  
Neonatal Rat ◽  
Interleukin 4 ◽  
Neonatal Rats ◽  
Alveolar Cells ◽  
Rsv Infection ◽  
Ifn Γ ◽  
Syncytial Virus

Purpose: To investigate the effect of chlorobenzoxime on respiratory syncytial virus (RSV) infection in vitro in lung alveolar cells and in vivo in neonatal rats, as well as the mechanism of action involved. Methods: RSV infection in neonatal rats was induced via intranasal administration of 2 x 106PFU viral particles. Reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting were used for determination of changes in interleukin expression. Results: RSV infection in BEAS-2B cells caused significant reduction in viability and marked alteration in morphological appearance (p < 0.05). Exposure of RSV-infected BEAS-2B cells to chlorobenzoxime prevented viability reduction and changes in morphology, and led to reductions in RSV-mediated increases in levels of interleukin-6 and interleukin-8. Moreover, RSV infection significantly enhanced ROS levels in BEAS-2B cells, when compared to control cells (p < 0.05). Chlorobenzoxime at a concentration of 30 μM completely suppressed RSV-mediated generation of ROS in BEAS-2B cells. In neonatal rats, RSV-induced upregulation of interleukin-4, interleukin-13 and TNF-α, were suppressed in bronchoalveolar lavage fluid (BALF) and lung tissues by chlorobenzoxime. Moreover, the RSVmediated reduction in IFN-γ was maximally blocked by chlorobenzoxime at a dose of 10 mg/mL. Chlorobenzoxime enhanced the proportion of IFN-γ -producing cells in neonatal rat BALF. Conclusion: Chlorobenzoxime exhibits antiviral against RSV infection in neonatal rats via increase in dendritic cell population, leading to inhibition of cytokine production. Therefore, chlorobenzoxime is a potential therapeutic agent for RSV infection. Keywords: Respiratory syncytial virus, Cytokines, Dendritic cells, Lung aveolar cells, Morphology, Interleukins

Regulation of calcium homeostasis in the fetal and neonatal rat

1981 ◽  
Vol 240 (4) ◽  
pp. E367-E372 ◽  
Author(s):  
M. L. Thomas ◽  
C. S. Anast ◽  
L. R. Forte
Keyword(s):  
Parathyroid Hormone ◽  
Cyclic Amp ◽  
Phosphodiesterase Inhibitor ◽  
Mean Value ◽  
Neonatal Rat ◽  
Neonatal Rats ◽  
Newborn Rats ◽  
Circulating Levels ◽  
Immunoreactive Parathyroid Hormone

Circulating levels of calcium (Ca) and immunoreactive parathyroid hormone (IPTH), and the renal cyclic AMP responses to PTH, calcitonin (CT), and vasopressin (VP) were measured in fetal and neonatal rats. Serum Ca increased from a mean value of 9.1 mg/dl on the 19th day of gestation to 10.9 on day 20. Circulating IPTH decreased from 875 pg/ml to 213. Serum Ca declined rapidly after birth to a nadir of 7.6 by 3 h and IPTH increased to 2,006 pg/ml, indicating that fetal and newborn parathyroids are capable of responding appropriately to changes in circulating Ca. Renal responsiveness to hormones was assessed in vitro in the presence of methylisobutylxanthine, a phosphodiesterase inhibitor. The tissue cyclic AMP response to PTH and CT (15- to 18-fold over basal) was greatest at gestational days 18 and 19, progressively declined throughout the remainder of gestation, and remained low during the first 24 h after birth (6- to 7-fold). Renal cyclic AMP response to VP remained consistently low throughout this period. The depressed renal cyclic AMP response to PTH at the time of birth may contribute to the hypocalcemia found in newborn rats.

scholarly journals Lactobacillus rhamnosusGG Suppresses MeningiticE. coliK1 Penetration across Human Intestinal Epithelial Cells In Vitro and Protects Neonatal Rats against Experimental Hematogenous Meningitis

2009 ◽  
Vol 2009 ◽  
pp. 1-7 ◽  
Author(s):  
Sheng-He Huang ◽  
Lina He ◽  
Yanhong Zhou ◽  
Chun-Hua Wu ◽  
Ambrose Jong
Keyword(s):  
Neonatal Sepsis ◽  
Lactobacillus Rhamnosus ◽  
Inhibitory Effect ◽  
Neonatal Rat ◽  
Control Group ◽  
Dependent Manner ◽  
Neonatal Rats ◽  
E Coli

The purpose of this study was to examine prophylactic efficacy of probiotics in neonatal sepsis and meningitis caused byE. coliK1. The potential inhibitory effect ofLactobacillus rhamnosusGG (LGG) on meningiticE. coliK1 infection was examined by using (i) in vitro inhibition assays with E44 (a CSF isolate from a newborn baby withE. colimeningitis), and (ii) the neonatal rat model ofE. colisepsis and meningitis. The in vitro studies demonstrated that LGG blocked E44 adhesion, invasion, and transcytosis in a dose-dependent manner. A significant reduction in the levels of pathogen colonization,E. colibacteremia, and meningitis was observed in the LGG-treated neonatal rats, as assessed by viable cultures, compared to the levels in the control group. In conclusion, probiotic LGG strongly suppresses meningiticE. colipathogens in vitro and in vivo. The results support the use of probiotic strains such as LGG for prophylaxis of neonatal sepsis and meningitis.

scholarly journals Nicotinic Agonist Inhibits Cardiomyocyte Apoptosis in CVB3-Induced Myocarditis via α3β4-nAChR/PI3K/Akt-Dependent Survivin Upregulation

2019 ◽  
Vol 2019 ◽  
pp. 1-13
Author(s):  
Ping Li ◽  
Yaoyao Yan ◽  
Youyang Shi ◽  
Bo Cheng ◽  
Yi Zhan ◽  
...  
Keyword(s):  
Caspase 3 ◽  
Viral Myocarditis ◽  
Cardiomyocyte Apoptosis ◽  
Neonatal Rat ◽  
Akt Pathway ◽  
Antiapoptotic Protein ◽  
Protein Expressions ◽  
Induced Apoptosis ◽  
Nicotinic Agonist

Background. Cardiomyocyte apoptosis is critical for the development of coxsackievirus B3- (CVB3-) induced myocarditis, which is a common cardiac disease that may result in heart failure or even sudden death. Previous studies have associated CVB3-induced apoptosis with the downregulation of antiapoptotic proteins. Here, attempts were made to examine whether nicotinic acetylcholine receptors (nAChRs), especially α3β4-nAChRs, were a novel therapeutic antiapoptotic target via the activation of survivin, a strong antiapoptotic protein, in viral myocarditis (VMC). Methods and Results. In the present study, we demonstrated that nAChRs, α3β4-nAChR subunits in particular, were present and upregulated in CVB3-infected neonatal rat cardiomyocytes (NRC) and H9c2 cells by RT-qPCR. The function of α3β4-nAChRs was next examined using its specific blocker α-CTX AuIB in vitro. The results of the TUNEL assay and western blot experiments showed that the block of α3β4-nAChRs abrogated nicotine-mediated protection of NRC from CVB3-induced apoptosis, and this effect displayed a substantial correlation with the protein expressions of pAkt, survivin, and Cleaved Caspase-3. Hence, the involvement of the PI3K/Akt pathway was further verified by LY294002, a selective inhibitor of PI3K. As a result, nicotine-mediated induction of pAkt and survivin was abolished by LY294002; meanwhile, apoptotic NRC were increased accompanied by an increase of Cleaved Caspase-3 expression. Regarding CVB3-infected BALB/c mice, the α-CTX AuIB- and LY294002-treated groups had a lower survival rate, deteriorative ventricular systolic function, and more severe inflammation than the nicotine-treated group and the modulation of pAkt, survivin, and Cleaved Caspase-3 protein expressions was similar to that in CVB3-infected NRC. In addition, we found that a nicotinic agonist reduced CVB3 replication in a dose-dependent manner in vitro, which indicates that nAChR activation may serve as a possible protection mechanism of CVB3-induced myocarditis. Conclusions. Our study demonstrated that α3β4-nAChR subunits are essential in the nicotine-mediated antiapoptotic effect of protecting cardiomyocytes from CVB3-induced apoptosis in vivo and in vitro. This protection correlated with the PI3K/Akt pathway and the inducement of the antiapoptotic protein survivin. A combination of these mechanisms serves as a novel protective response to treat viral myocarditis.

scholarly journals Protective effect of midazolam against convulsion in neonatal rats via down-regulation of LC3 and Beclin-1 expression

2020 ◽  
Vol 19 (3) ◽  
pp. 557-563
Author(s):  
Juan Shi ◽  
Shuzhong Jiang ◽  
Qinhua Wang ◽  
Jiajia Hua ◽  
Feifan Xu ◽  
...  
Keyword(s):  
Lactate Dehydrogenase ◽  
Caspase 3 ◽  
Mitochondrial Pathway ◽  
Beclin 1 ◽  
Neonatal Rats ◽  
Caspase 9 ◽  
Protein Expressions ◽  
Brain Water ◽  
Inhibitor Treatment

Purpose: To investigate the effect of midazolam on growth of neurocytes in vitro and in neonatal rats. Methods: Neurocyte proliferation and activity of lactate dehydrogenase were assessed by MTT and lactate dehydrogenase assays, respectively. Western blotting was used to determine the effect of midazolam on LC3, Bax, p62 and Beclin-1 protein expressions. Results: The suppression of neurocyte proliferation byconvulsion was alleviated significantly (p < 0.05) by midazolum treatment. Exposure of convulsion model of neurocytes to midazolum suppressed LC3, Bax, p62 and Beclin-1 protein expression. Midazolum exposure of convulsion model of neurocytes suppressed LDH, caspase-3, caspase-8 and caspase-9 activities. The 3-MA (autophagy inhibitor) treatment also significantly (p < 0.05) promoted neurocyte viability after convulsion induction. In convulsion-induced neurocytes, 3-MA exposure suppressed expression of caspase-3/8/9, LC3, Bax, Beclin-1 and p62, while application of midazolum treatment to the rats with convulsion markedly decreased brain water content and neurocyte apoptosis (p < 0.05). Treatment with midazolum inhibited LC3, p62 and Beclin-1 expression in the rat model of convulsion. Conclusion: Midazolum promotes neurocyte proliferation and inhibits edema development via downregulation of autophagy. Therefore, midazolum can potentially be used for the treatment of convulsion, but further studies need to be carried out first. Keywords: Convulsion, Neurocytes, Caspase, Autophagy, Mitochondrial pathway

Sulfide-induced perturbations of the neuronal mechanisms controlling breathing in rats

1995 ◽  
Vol 78 (2) ◽  
pp. 433-440 ◽  
Author(s):  
J. J. Greer ◽  
R. J. Reiffenstein ◽  
A. F. Almeida ◽  
J. E. Carter
Keyword(s):  
Respiratory Rhythm ◽  
Intraperitoneal Administration ◽  
Experimental Models ◽  
Neonatal Rat ◽  
Ventrolateral Medulla ◽  
Neonatal Rats ◽  
Breathing Patterns ◽  
Adult Rats

The effects of sulfide on neonatal rat respiration were studied. Two in vitro experimental models were utilized: the isolated brain stem-spinal cord preparation and the medullary slice preparation containing respiratory rhythm-generating regions from neonatal rats. Plethysmographic measurements of the effects of sulfide on the breathing patterns of unanesthetized neonatal rats were also made to compare the sensitivities of neonatal and adult rats to sulfide toxicity. In vitro, sulfide acted at sites within the ventrolateral medulla to depress the frequency of respiratory rhythmic discharge by approximately 50–60%. However, the neuronal network underlying respiratory rhythmogenesis continued to function in the presence of concentrations of sulfide far beyond those deemed to be lethal in vivo. Intraperitoneal administration of sulfide caused a dose-dependent decrease in the frequency and amplitude of breathing of neonatal rats of all ages (0–19 days postnatal), although the sensitivity to sulfide increased with age. We hypothesize that the rapid suppression of breathing caused by sulfide is due to changes in neuronal excitability within respiratory rhythm-generating centers rather than, as previously hypothesized, to perturbations of cellular oxidative metabolism.

scholarly journals VIP activates primordial follicles of rat through ERK-mTOR pathway in tissue culture

Reproduction ◽  
2019 ◽  
Vol 157 (5) ◽  
pp. 475-484 ◽  
Author(s):  
Song Li ◽  
Qi Fan ◽  
Yanqiu Xie ◽  
Haiyan Lin ◽  
Qi Qiu ◽  
...  
Keyword(s):  
Primordial Follicle ◽  
Reproductive Technologies ◽  
Treated Group ◽  
Signalling Pathway ◽  
Neonatal Rat ◽  
Granular Cells ◽  
Primordial Follicles ◽  
Mtor Signalling ◽  
Mtor Signalling Pathway

In vitro activation of primordial follicles is becoming more essential in assisted reproductive technologies. Vasoactive intestinal peptide (VIP) is one of the members of the neurotrophin family which has demonstrated to have an impact on follicle development in recent years. This study aims to investigate the effect of VIP on the activation of primordial follicles in neonatal rat in an in vitro culture system and to determine the relevant molecular mechanism of their activation. Ovaries of 4-day-old rats were examined for the expression of VIP receptors and were cultured in mediums containing VIP with or without inhibitors of the ERK–mTOR signalling pathway. They were then collected for histological analysis or measurement of the molecular expression of this pathway. The receptors of VIP were found in granular cells and oocytes of primordial and early-growing follicles in neonatal ovary. The ratio of growing follicle increased in the presence VIP at different concentrations, with the highest level of increase being observed in the 10−7 mol/L VIP-treated group. The ratio of PCNA-positive granular cells was also increased, while that of the apoptotic oocytes were decreased, and protein analysis showed increased phosphorylation of ERK1/2, mTOR and RPS6 in the VIP-treated group. However, the effect of VIP on the activation of primordial follicle became insignificant with the addition of MEK inhibitor (U0126) or mTORC1 inhibitor (rapamycin). This study indicated that VIP could activate neonatal rat primordial follicle through the ERK-mTOR signalling pathway, suggesting a strategy for in vitro primordial follicle recruitment.

scholarly journals Simvastatin Attenuates Neurogenetic Damage and Improves Neurocongnitive Deficits Induced by Isoflurane in Neonatal Rats

2018 ◽  
Vol 46 (2) ◽  
pp. 618-632 ◽  
Author(s):  
Ning Wang ◽  
Yang Lu ◽  
Kui Wang ◽  
Wei-song Li ◽  
Pan Lu ◽  
...  
Keyword(s):  
Cognitive Deficits ◽  
Caspase 3 ◽  
Morris Water Maze Test ◽  
Neonatal Rats ◽  
Neurocognitive Deficits ◽  
Wide Range ◽  
Protein Expressions ◽  
Gsk 3Β

Background/Aims: Isoflurane inhibited neurogenesis and induced subsequent neurocognitive deficits in developing brain. Simvastatin exerts neuroprotection in a wide range of brain injury models. In the present study, we investigated whether simvastatin could attenuate neurogenetic inhibition and cognitive deficits induced by isoflurane exposure in neonatal rats. Methods: Sprague-Dawley rats at postnatal day (PND) 7 and neural stem cells (NSCs) were treated with either gas mixture, isoflurane, or simvastatin 60 min prior to isoflurane exposure, respectively. The rats were decapitated at PND 8 and PND 10 for detection of neurogenesis in the subventricular zone (SVZ) and subgranular zone (SGZ) of the hippocampus by immunostaining. NSC proliferation, viability and apoptosis were assessed by immunohistochemistry, CCK-8 and TUNEL, respectively. The protein expressions of caspase-3, p-Akt and p-GSK-3β both in vivo and vitro were assessed by western blotting. Cognitive functions were assessed by Morris Water Maze test and context fear conditioning test at the adult. Results: Isoflurane exposure inhibited neurogenesis in the SVZ and SGZ, decreased NSC proliferation and viability, promoted NSC apoptosis and led to late cognitive deficits. Furthermore, isoflurane increased caspase-3 expression and decreased protein expressions of p-Akt and p-GSK-3β both in vivo and in vitro. Pretreatment with simvastatin attenuated isoflurane-elicited changes in NSCs and cognitive function. Co-treatment with LY294002 reversed the effect of simvastatin on NSCs in vitro. Conclusion: We for the first time showed that simvastatin, by upregulating Akt/GSK-3β signaling pathway, alleviated isoflurane-induced neurogenetic damage and neurocognitive deficits in developing rat brain.

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