scholarly journals Proteomics and functional study reveal kallikrein-6 enhances communicating hydrocephalus

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Lei Yuan ◽  
Dongdong Zou ◽  
Xia Yang ◽  
Xin Chen ◽  
Youming Lu ◽  
...  
Keyword(s):  
Rat Model ◽  
Differentially Expressed ◽  
Functional Study ◽  
Control Group ◽  
Communicating Hydrocephalus ◽  
Loss Of Function ◽  
Neuronal Nuclei ◽  
Hoechst Staining ◽  
Interfering Rna ◽  
Kallikrein 6

Abstract Background Communicating hydrocephalus (CH) is a common neurological disorder caused by a blockage of cerebrospinal fluid. In this study, we aimed to explore the potential molecular mechanism underlying CH development. Methods Quantitative proteomic analysis was performed to screen the differentially expressed proteins (DEPs) between patients with and without CH. A CH rat model was verified by Hoechst staining, and the co-localization of the target protein and neuron was detected using immunofluorescence staining. Loss-of-function experiments were performed to examine the effect of KLK6 on the synapse structure. Results A total of 11 DEPs were identified, and kallikrein 6 (KLK6) expression was found to be significantly upregulated in patients with CH compared with that in patients without CH. The CH rat model was successfully constructed, and KLK6 was found to be co-localized with neuronal nuclei in brain tissue. The expression level of IL-1β, TNF-α, and KLK6 in the CH group was higher than that in the control group. After knockdown of KLK6 expression using small-interfering RNA (siRNA), the expression levels of synapsin-1 and PSD95 in neuronal cells were increased, and the length, number, and structure of synapses were significantly improved. Following siRNA interference KLK6 expression, 5681 differentially expressed genes (DEGs) were identified in transcriptome profile. The upregulated DEGs of Appl2, Nav2, and Nrn1 may be involved in the recovery of synaptic structures after the interference of KLK6 expression. Conclusions Collectively, KLK6 participates in the development of CH and might provide a new target for CH treatment.

scholarly journals Proteomic and Functional Analyses Reveal That Kallikrein 6 Enhances Communicating Hydrocephalus by Injuring Neuronal Synapses

2021 ◽  
Author(s):  
Lei Yuan ◽  
Dongdong Zou ◽  
Xia Yang ◽  
Xin Chen ◽  
Youming Lu ◽  
...  
Keyword(s):  
Rat Model ◽  
Differentially Expressed ◽  
Control Group ◽  
Communicating Hydrocephalus ◽  
Loss Of Function ◽  
Neuronal Nuclei ◽  
Neuronal Synapses ◽  
Hoechst Staining ◽  
Synaptic Structures ◽  
Kallikrein 6

Abstract Communicating hydrocephalus (CH) is a common neurological disorder caused by a blockage of cerebrospinal fluid. In this study, we aimed to explore the potential molecular mechanism underlying CH development. Quantitative proteomic analysis was performed to screen the differentially expressed proteins (DEPs) between patients with and without CH. A CH rat model was verified by Hoechst staining, and the co-localization of the target protein and neuron was detected using immunofluorescence staining. Loss-of-function experiments were performed to examine the effect of KLK6 on the synapse structure. A total of 11 DEPs were identified, and kallikrein 6 (KLK6) expression was found to be significantly upregulated in patients with CH compared with that in patients without CH. The CH rat model was successfully constructed, and KLK6 was found to be co-localized with neuronal nuclei in brain tissue. The expression level of KLK6 in the CH group was higher than that in the control group. After interference of KLK6 expression, the expression levels of synapsin-1 and PSD95 in neuronal cells were increased, and the length, number, and structure of synapses were significantly improved. The transcriptome profile (PRJNA719985) after interference of KLK6 expression was obtained, and 5,681 differentially expressed genes (DEGs) were identified. The upregulated DEGs of Appl2, Nav2, and Nrn1 may be involved in the recovery of synaptic structures after interference of KLK6 expression. Collectively, KLK6 participates in the development of CH and might provide a new target for CH treatment.

scholarly journals Functional study of a N-terminal CPVT mutation RyR2R420Q in patient specific hiPSC-CMs model

EP Europace ◽  
2021 ◽  
Vol 23 (Supplement_3) ◽  
Author(s):  
L Yin ◽  
P Joanne ◽  
R Perrier ◽  
P Gerbaud ◽  
P Lechene ◽  
...  
Keyword(s):  
Induced Pluripotent Stem Cell ◽  
Calcium Transient ◽  
Polymorphic Ventricular Tachycardia ◽  
Functional Study ◽  
Patient Specific ◽  
Control Group ◽  
Structural Abnormality ◽  
Loss Of Function ◽  
Adrenergic Stimulation ◽  
Gain Of Function

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): ANR (Agence Nationale de la Rercherche) Catecholaminergic polymorphic ventricular tachycardia (CPVT) is a lethal genetic arrhythmia that manifests by syncope or sudden death in children and young adults under stress conditions without obvious cardiac structural abnormality. A novel CPVT mutation located in the RyR2 N terminal portion has been identified in a Spanish family (RyR2R420Q). According to the studies of RyR2 function in HEK293 cell line, this mutation presented gain of function at low cytosolic intracellular Ca2+ concentration ([Ca2+]i) and loss of function at high [Ca2+]i. Moreover, KI mice heterozygous for this mutation presented bradycardia and sino-atrial node (SAN) dysfunction. Here we generated induced pluripotent stem cell (hiPSC) from two brothers (one with mutation, the other without mutation as control) of this family and differentiated them into cardiomyocytes (hiPSC-CM). In order to verify that the differentiated cells were well cardiomyocytes, we did immunofluorescence labelling to detect the α-actinin expression and found that around 90% cells were α-actinin positive in both groups of hiPS-CMs. Then the calcium transient was studied by confocal microscopy and the action potential (AP) by micro-electrode technique. The characteristics of spontaneous AP of mutated cells were mostly similar to that of control cells, but more mutated cells presented proarrhythmic behaviors under adrenergic stimulation. hiPSC-CM are immature cardiomyocytes and contract spontaneously. In order to be able to analyze [Ca2+]i transient characteristics, we paced the cells at a constant rate of 1 Hz by field stimulation through two Pt electrodes. Sarcoplasmic reticulum (SR) Ca2+ load was estimated by rapid caffeine (10 mM) application. hiPSC-CMs from the RyR2R420Q carrier presented smaller SR Ca2+ load than those from the control person, whereas their fractional release (the [Ca2+]i transient normalized by the amount of Ca2+ stored in the SR) was higher than that in control group, indicating a gain-of-function mutation.  Even if SR Ca2+ load was smaller in RyR2R420Q cells, they often presented proarrhythmogenic behavior such as Ca2+ waves. The fact was further enhanced during β-adrenergic stimulation, pointing to this model as a valuable tool to study the CPVT disease in human cells.

scholarly journals Restorative potential of (−)-epicatechin in a rat model of Gulf War illness muscle atrophy and fatigue

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Israel Ramirez-Sanchez ◽  
Viridiana Navarrete-Yañez ◽  
Alejandra Garate-Carrillo ◽  
Modesto Lara-Hernandez ◽  
Judith Espinosa-Raya ◽  
...  
Keyword(s):  
Muscle Atrophy ◽  
Rat Model ◽  
Inflammatory Cytokine ◽  
Gulf War ◽  
Plasma Metabolites ◽  
Control Group ◽  
Loss Of Function ◽  
Gulf War Illness ◽  
Mediators Of Inflammation ◽  
Male Wistar Rats

AbstractWe examined in a rat model of Gulf War illness (GWI), the potential of (−)-epicatechin (Epi) to reverse skeletal muscle (SkM) atrophy and dysfunction, decrease mediators of inflammation and normalize metabolic perturbations. Male Wistar rats (n = 15) were provided orally with pyridostigmine bromide (PB) 1.3 mg/kg/day, permethrin (PM) 0.13 mg/kg/day (skin), DEET 40 mg/kg/day (skin) and were physically restrained for 5 min/day for 3 weeks. A one-week period ensued to fully develop the GWI-like profile followed by 2 weeks of either Epi treatment at 1 mg/kg/day by gavage (n = 8) or water (n = 7) for controls. A normal, control group (n = 15) was given vehicle and not restrained. At 6 weeks, animals were subjected to treadmill and limb strength testing followed by euthanasia. SkM and blood sampling was used for histological, biochemical and plasma pro-inflammatory cytokine and metabolomics assessments. GWI animals developed an intoxication profile characterized SkM atrophy and loss of function accompanied by increases in modulators of muscle atrophy, degradation markers and plasma pro-inflammatory cytokine levels. Treatment of GWI animals with Epi yielded either a significant partial or full normalization of the above stated indicators relative to normal controls. Plasma metabolomics revealed that metabolites linked to inflammation and SkM waste pathways were dysregulated in the GWI group whereas Epi, attenuated such changes. In conclusion, in a rat model of GWI, Epi partially reverses detrimental changes in SkM structure including modulators of atrophy, inflammation and select plasma metabolites yielding improved function.

scholarly journals Restorative Effects of (-)-Epicatechin in a Rat Model of Gulf War Illness Muscle Atrophy and Fatigue

2021 ◽  
Author(s):  
Israel Ramirez-Sanchez ◽  
Viridiana Navarrrete ◽  
Alejandra Garate-Carrillo ◽  
Modesto Lara-Hernandez ◽  
Judith Espinosa-Raya ◽  
...  
Keyword(s):  
Muscle Atrophy ◽  
Rat Model ◽  
Inflammatory Cytokine ◽  
Gulf War ◽  
Plasma Metabolites ◽  
Control Group ◽  
Loss Of Function ◽  
Gulf War Illness ◽  
Mediators Of Inflammation ◽  
Male Wistar Rats

Abstract We examined in a rat model of Gulf War illness (GWI), the potential of (-)-epicatechin (Epi) to reverse skeletal muscle (SkM) atrophy and dysfunction, decrease mediators of inflammation and normalize metabolic perturbations. Male Wistar rats (n = 15) were provided orally with pyridostigmine bromide (PB) 1.3 mg/kg/day, permethrin (PM) 0.13 mg/kg/day (skin), DEET 40 mg/kg/day (skin) and were physically restrained for 5 min/day for 3 weeks. A one-week period ensued to fully develop the GWI-like profile followed by 2 weeks of either Epi treatment at 1 mg/kg/day by gavage (n = 8) or water (n = 7) for controls. A normal, control group (n = 15) was given vehicles and not restrained. At 6 weeks, animals were subjected to treadmill and limb strength testing followed by euthanasia. SkM and blood sampling was used for histological, biochemical and plasma pro-inflammatory cytokine and metabolomics assessments. GWI animals developed an intoxication profile characterized SkM atrophy and loss of function accompanied by increases in modulators of muscle atrophy, degradation markers and plasma pro-inflammatory cytokine levels. Treatment of GWI animals with Epi yielded either a significant partial or full normalization of the above stated indicators relative to normal controls. Plasma metabolomics revealed that metabolites linked to inflammation and SkM waste pathways were dysregulated in the GWI group whereas Epi, attenuated such changes. In conclusion, in a rat model of GWI, Epi reverses detrimental changes in SkM structure including modulators of atrophy, inflammation and select plasma metabolites yielding improved function.

Bioinformatics Analysis and Validation of Differentially Expressed MicroRNAs with Their Target Genes Involved in GLP-1RA Facilitated Osteogenesis

2020 ◽  
Vol 15 ◽  
Author(s):  
Na Wang ◽  
Yukun Li ◽  
Sijing Liu ◽  
Liu Gao ◽  
Chang Liu ◽  
...  
Keyword(s):  
High Throughput Sequencing ◽  
Target Genes ◽  
Bioinformatics Analysis ◽  
Differentially Expressed ◽  
Functional Study ◽  
Regulation Network ◽  
Glucagon Like Peptide 1 ◽  
G Protein Coupled ◽  
Lower Expression

Background: Recent studies revealed that the hypoglycemic hormone, glucagon-like peptide-1 (GLP-1), acted as an important modulator in osteogenesis of bone marrow derived mesenchymal stem cells (BMSCs). Objectives: The aim of this study was to identify the specific microRNA (miRNA) using bioinformatics analysis and validate the presence of differentially expressed microRNAs with their target genes after GLP-1 receptor agonist (GLP-1RA) administration involved in ostogenesis of BMSCs. Methods: MiRNAs were extracted from BMSCs after 5 days’ treatment and sent for high-throughput sequencing for differentially expressed (DE) miRNAs analyses. Then the expression of the DE miRNAs verified by the real-time RT-PCR analyses. Target genes were predicted, and highly enriched GOs and KEGG pathway analysis were conducted using bioinformatics analysis. For the functional study, two of the target genes, SRY (sex determining region Y)-box 5 (SOX5) and G protein-coupled receptor 84 (GPR84), were identified. Results: A total of 5 miRNAs (miRNA-509-5p, miRNA-547-3p, miRNA-201-3p, miRNA-201-5p, and miRNA-novel-272-mature) were identified differentially expressed among groups. The expression of miRNA-novel-272-mature were decreased during the osteogenic differentiation of BMSCs, and GLP-1RA further decreased its expression. MiRNA-novel-272-mature might interact with its target mRNAs to enhance osteogenesis. The lower expression of miRNA-novel-272-mature led to an increase in SOX5 and a decrease in GPR84 mRNA expression, respectively. Conclusions: Taken together, these results provide further insights to the pharmacological properties of GLP-1RA and expand our knowledge on the role of miRNAs-mRNAs regulation network in BMSCs’ differentiation.

scholarly journals Salidroside promoted osteogenic differentiation of adipose-derived stromal cells through Wnt/β-catenin signaling pathway

2021 ◽  
Vol 16 (1) ◽  
Author(s):  
Xiao-hua Li ◽  
Fu-ling Chen ◽  
Hong-lin Shen
Keyword(s):  
Western Blot ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Differentially Expressed Genes ◽  
Stromal Cells ◽  
Differentially Expressed ◽  
Calcium Deposition ◽  
Western Blot Assay ◽  
Adipose Derived Stromal Cells ◽  
Interfering Rna

Abstract Background Bone disease causes short-term or long-term physical pain and disability. It is necessary to explore new drug for bone-related disease. This study aimed to explore the role and mechanism of Salidroside in promoting osteogenic differentiation of adipose-derived stromal cells (ADSCs). Methods ADSCs were isolated and treated with different dose of Salidroside. Cell count kit-8 (CCK-8) assay was performed to assess the cell viability of ADSCs. Then, ALP and ARS staining were conducted to assess the early and late osteogenic capacity of ADSCs, respectively. Then, differentially expressed genes were obtained by R software. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the differentially expressed genes were further analyzed. The expression of OCN, COL1A1, RUNX2, WNT3A, and β-catenin were measured by real-time PCR and Western blot analysis. Last, β-catenin was silenced by small interfering RNA. Results Salidroside significantly increased the ADSCs viability at a dose-response manner. Moreover, Salidroside enhanced osteogenic capacity of ADSCs, which are identified by enhanced ALP activity and calcium deposition. A total of 543 differentially expressed genes were identified between normal and Salidroside-treated ADSCs. Among these differentially expressed genes, 345 genes were upregulated and 198 genes were downregulated. Differentially expressed genes enriched in the Wnt/β-catenin signaling pathway. Western blot assay indicated that Salidroside enhanced the WNT3A and β-catenin expression. Silencing β-catenin partially reversed the promotion effects of Salidroside. PCR and Western blot results further confirmed these results. Conclusion Salidroside promoted osteogenic differentiation of ADSCs through Wnt/β-catenin signaling pathway.

scholarly journals Analysis of Circulating microRNA Signatures and Preeclampsia Development

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1003
Author(s):  
Margarita L. Martinez-Fierro ◽  
Idalia Garza-Veloz
Keyword(s):  
Pregnant Women ◽  
Expression Profiling ◽  
Gene Prediction ◽  
Microrna Expression ◽  
Differentially Expressed ◽  
Control Group ◽  
Case Group ◽  
Circulating Microrna ◽  
Activated T Cells ◽  
Serum Microrna

microRNAs are important regulators of cell processes and have been proposed as potential preeclampsia biomarkers. We evaluated serum microRNA expression profiling to identify microRNAs involved in preeclampsia development. Serum microRNA expression profiling was evaluated at 12, 16, and 20 weeks of gestation (WG), and at the time of preeclampsia diagnosis. Two groups were evaluated using TaqMan low-density array plates: a control group with 18 normotensive pregnant women and a case group with 16 patients who developed preeclampsia during the follow-up period. Fifty-three circulating microRNAs were differentially expressed between groups (p < 0.05). Compared with controls, hsa-miR-628-3p showed the highest relative quantity values (at 12 WG = 7.7 and at 20 WG = 3.45) and the hsa-miRs -151a-3p and -573 remained differentially expressed from 16 to 20 WG (p < 0.05). Signaling pathways including cancer-related, axon guidance, Neurotrophin, GnRH, VEGF, and B/T cell receptor, were most commonly altered. Further target gene prediction revealed that nuclear factor of activated T-cells 5 gene was included among the transcriptional targets of preeclampsia-modulated microRNAs. Specific microRNAs including hsa-miRs -628-3p, -151a-3p, and -573 were differentially expressed in serum of pregnant women before they developed preeclampsia compared with controls and their participation in the preeclampsia development should be considered.

scholarly journals Systemic Administration of G-CSF Accelerates Bone Regeneration and Modulates Mobilization of Progenitor Cells in a Rat Model of Distraction Osteogenesis

2021 ◽  
Vol 22 (7) ◽  
pp. 3505
Author(s):  
Flavy Roseren ◽  
Martine Pithioux ◽  
Stéphane Robert ◽  
Laure Balasse ◽  
Benjamin Guillet ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Progenitor Cells ◽  
Distraction Osteogenesis ◽  
Rat Model ◽  
Late Phase ◽  
Control Group ◽  
Bone Lengthening ◽  
Hematopoietic Stem ◽  
Osteoblast Activity ◽  
Positron Emission

Granulocyte colony-stimulating factor (G-CSF) was shown to promote bone regeneration and mobilization of vascular and osteogenic progenitor cells. In this study, we investigated the effects of a systemic low dose of G-CSF on both bone consolidation and mobilization of hematopoietic stem/progenitor cells (HSPCs), endothelial progenitor cells (EPCs) and mesenchymal stromal cells (MSCs) in a rat model of distraction osteogenesis (DO). Neovascularization and mineralization were longitudinally monitored using positron emission tomography and planar scintigraphy. Histological analysis was performed and the number of circulating HSPCs, EPCs and MSCs was studied by flow cytometry. Contrary to control group, in the early phase of consolidation, a bony bridge with lower osteoclast activity and a trend of an increase in osteoblast activity were observed in the distracted callus in the G-CSF group, whereas, at the late phase of consolidation, a significantly lower neovascularization was observed. While no difference was observed in the number of circulating EPCs between control and G-CSF groups, the number of MSCs was significantly lower at the end of the latency phase and that of HSPCs was significantly higher 4 days after the bone lengthening. Our results indicate that G-CSF accelerates bone regeneration and modulates mobilization of progenitor cells during DO.

scholarly journals CircHIPK3 regulates the autophagy and apoptosis of hypoxia/reoxygenation-stimulated cardiomyocytes via the miR-20b-5p/ATG7 axis

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Zhimei Qiu ◽  
Yan Wang ◽  
Weiwei Liu ◽  
Chaofu Li ◽  
Ranzun Zhao ◽  
...  
Keyword(s):  
Cell Injury ◽  
Ischemia Reperfusion ◽  
Circular Rna ◽  
Control Group ◽  
Loss Of Function ◽  
Cardiomyocyte Autophagy ◽  
Myocardial Ischemia Reperfusion ◽  
Injury Research ◽  
Hypoxia Reoxygenation

AbstractAutophagy and apoptosis are involved in myocardial ischemia/reperfusion (I/R) injury. Research indicates that circular RNA HIPK3 (circHIPK3) is crucial to cell autophagy and apoptosis in various cancer types. However, the role of circHIPK3 in the regulation of cardiomyocyte autophagy and apoptosis during I/R remains unknown. Our study aimed to examine the regulatory effect of circHIPK3 during myocardial I/R and investigate its mechanism in cardiomyocyte autophagy and apoptosis. Methods and results. The expression of circHIPK3 was upregulated during myocardial I/R injury and hypoxia/reoxygenation (H/R) injury of cardiomyocytes. To study the potential role of circHIPK3 in myocardial H/R injury, we performed gain-of-function and loss-of-function analyses of circHIPK3 in cardiomyocytes. Overexpression of circHIPK3 significantly promoted H/R-induced cardiomyocyte autophagy and cell injury (increased intracellular reactive oxygen species (ROS) and apoptosis) compared to those in the control group, while silencing of circHIPK3 showed the opposite effect. Further research found that circHIPK3 acted as an endogenous miR-20b-5p sponge to sequester and inhibit miR-20b-5p activity, resulting in increased ATG7 expression. In addition, miR-20b-5p inhibitors reversed the decrease in ATG7 induced by silencing circHIPK3. Conclusions. CircHIPK3 can accelerate cardiomyocyte autophagy and apoptosis during myocardial I/R injury through the miR-20b-5p/ATG7 axis. These data suggest that circHIPK3 may serve as a potential therapeutic target for I/R.

scholarly journals Target identification of hepatic fibrosis using Pien Tze Huang based on mRNA and lncRNA

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jinhang Zhu ◽  
Di Zhang ◽  
Ting Wang ◽  
Zhiliang Chen ◽  
Luan Chen ◽  
...  
Keyword(s):  
Liver Injury ◽  
Hepatic Fibrosis ◽  
Drug Targets ◽  
Enrichment Analysis ◽  
Inflammatory Cells ◽  
Differentially Expressed ◽  
Control Group ◽  
Traditional Chinese Herbal Medicine ◽  
Hepatic Diseases ◽  
Pien Tze Huang

AbstractHepatic fibrosis is a spontaneous wound-healing response triggered by chronic liver injury. Pien Tze Huang (PZH), a traditional Chinese herbal medicine, has been widely used to treat various hepatic diseases in Asia. We used a CCl4-induced mouse model to establish a PZH group of hepatic fibrosis mice treated with PZH and a control group of hepatic fibrosis mice without any treatment. We performed RNA-seq and mass spectrometry sequencing to investigate the mechanism of the PZH response in hepatic fibrosis and identified multiple differentially expressed transcripts (DETs) and proteins (DEPs) that may be drug targets of PZH. Liver functional indices, including serum albumin (ALB), alanine aminotransferase (ALT) and aspartate aminotransferase (AST), were significantly decreased in the PZH treatment group (P < 0.05) in the eighth week. Hematoxylin–eosin (HE), Masson and Sirius red staining demonstrated that PZH significantly inhibited infiltration of inflammatory cells and collagen deposition. A total of 928 transcripts and 138 proteins were differentially expressed in PZH-treated mice compared to the control group. Gene Ontology (GO) enrichment analysis suggested that PZH may alleviate liver injury and fibrosis by enhancing the immune process. Taken together, our results revealed that multiple DETs and DEPs may serve as drug targets of PZH in hepatic fibrosis patient in future clinical practice.

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