scholarly journals Do uterine PTGS2, PGFS, and PTGFR expression play a role in canine uterine inertia?

2021 ◽  
Author(s):  
Lea Magdalena Rempel ◽  
Karina Tietgen Andresen Lillevang ◽  
Ann-Kirstine thor Straten ◽  
Sólrún Barbara Friðriksdóttir ◽  
Hanna Körber ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Mrna Expression ◽  
Common Cause ◽  
Physiological Relevance ◽  
Uterine Inertia ◽  
Circular Layer ◽  
The Difference ◽  
Uterine Glands ◽  
Luminal Epithelial Cells

AbstractThe aetiology of primary uterine inertia (PUI), which is the most common cause of canine dystocia, is still not elucidated. Prostaglandins (PGs) play a crucial role in parturition. We hypothesized that the expression of prostaglandin endoperoxidase synthase 2 (PTGS2), PGF2α synthase (PGFS), and corresponding receptor (PTGFR) is altered in PUI. We investigated PTGS2, PGFS, and PTGFR mRNA expression, and PTGS2 and PGFS protein expression in interplacental (IP) and uteroplacental sites (UP) in bitches with PUI, obstructive dystocia (OD), and prepartum (PC). PTGS2, PGFS, and PTGFR mRNA expression did not differ significantly between PUI and OD (IP/UP). PTGFR ratio in UP was higher in PC than in OD (p = 0.014). PTGS2 immunopositivity was noted in foetal trophoblasts, luminal and superficial glandular epithelial cells, smooth muscle cells of both myometrial layers, and weakly and sporadically in deep uterine glands. PGFS was localized in luminal epithelial cells and in the epithelium of superficial uterine glands. PTGS2 and PGFS staining was similar between PUI and OD, while PGFS protein expression differed between OD and PC (p = 0.0215). For PTGS2, the longitudinal myometrial layer of IP stained significantly stronger than the circular layer, independent of groups. These results do not support a role for PTGS2, PGFS, and PTGFR in PUI. Reduced PGFS expression in IP during parturition compared with PC and the overall lack of placental PGFS expression confirm that PGFS is not the main source of prepartal PGF2alpha increase. The difference in PTGS2 expression between IP myometrial layers warrants further investigation into its physiological relevance.

scholarly journals AGR3 Regulates Airway Epithelial Junctions in Patients with Frequent Exacerbations of COPD

2021 ◽  
Vol 12 ◽  
Author(s):  
Rui Ye ◽  
Cuihong Wang ◽  
Pengbo Sun ◽  
Shuang Bai ◽  
Li Zhao
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Mrna Expression ◽  
Western Blotting ◽  
Human Lung ◽  
Airway Epithelial Cells ◽  
Copd Exacerbation ◽  
Airway Epithelial ◽  
Copd Exacerbations ◽  
E Cadherin

Background: The mechanisms underlying differences in the susceptibility to chronic obstructive pulmonary disease (COPD) exacerbations between patients are not well understood. Recent studies have shown that the patients with frequent COPD exacerbations is related to specific protein expression in lung tissue. Anterior gradient 3 (AGR3) is expressed in airway epithelial cells in the lung and proteomic analysis revealed that its expression is decreased in patients with frequent COPD exacerbations. Moreover, the loss of epithelial integrity might facilitate trans-epithelial permeability of pathogens in such patients. This study was performed to determine that AGR3 protein play a role in COPD frequency exacerbators.Methods: Human lung tissues were collected from current-smoking patients (Control; n = 15) as well as patients with infrequent COPD exacerbations (IFCOPD; n = 18) and frequent COPD exacerbations (FCOPD; n = 8). While AGR3 protein expression was measured by immunohistochemistry and western blotting, AGR mRNA expression was determined by real time quantitative polymerase chain reaction (RT-qPCR). Furthermore, adherent junctions (AJs) and tight junctions (TJs) protein expression in human lung tissues were measured by immunohistochemistry. The effects of cigarette smoke extract (CSE) on AJ and TJ protein and mRNA expression in BEAS-2B cells were assessed by western blotting and RT-qPCR. In addition, the effect of AGR3 overexpression and knockdown on AJ and TJ protein expression was determined.Results: AGR3 was mainly expressed in the airway epithelium and AGR3-positive products were localized in the cytoplasm. Western blotting and RT-qPCR results showed that AGR3 protein (p = 0.009) and mRNA (p = 0.04) expression in the FCOPD group was significantly lower than that in the IFCOPD group. Moreover, E-cadherin, occludin, and zonula occludens-1 (ZO-1) expression was lower in the FCOPD group than in the IFCOPD group. The protein and mRNA expression of E-cadherin, occludin, and ZO-1 was decreased within 24 h post-CSE exposure. AGR3 overexpression rescued CSE-induced downregulation of E-cadherin, occludin, and ZO-1.Conclusion: Difference in AGR3 expression in the lung tissue might be correlated with increased susceptibility to COPD exacerbation. AGR3 can prevent CSE-induced downregulation of E-cadherin, occludin, and ZO-1 in airway epithelial cells. Loss of AGR3 might promote viral and bacterial infection and induce immune inflammation to increase COPD exacerbation.

scholarly journals DOP87 SUCNR1 a novel key protagonist in fistula development

2020 ◽  
Vol 14 (Supplement_1) ◽  
pp. S126-S126
Author(s):  
C Bauset ◽  
L Gisbert-Ferrandiz ◽  
D Ortiz-Masia ◽  
S Coll ◽  
C Mamie ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Protein Expression ◽  
Mrna Expression ◽  
Intestinal Epithelial Cells ◽  
Fistula Formation ◽  
Fistula Tract ◽  
Intestinal Epithelial ◽  
Gene And Protein Expression ◽  
Emt Markers ◽  
E Cadherin

Abstract Background Intestinal fistula is a common complication in CD patients whose aetiology is still not well-characterised. It is associated with an exacerbated inflammation and epithelial-to-mesenchymal transition (EMT), a process which allows a switch from epithelial towards a fibrotic behaviour. We have recently reported that SUCNR1 mediates intestinal inflammation and fibrosis1 but its role in fistula has not yet been analysed. Therefore, we aim to analyse the role of SUCNR1 in EMT and in fistula formation. Methods Intestinal resections were obtained from CD and non-IBD patients. Fistula specimens were identified by the surgeons and collected from B3-CD patients. The expression of SUCNR1 and EMT markers was analysed by qPCR and the protein expression of SUCNR1 by immunohistochemistry. HT-29 cells were treated with succinate (0,0.1,0.5,1,5 mM) or TGF-β (5 ng/ml) during 48 h and transfected with SUCNR1 siRNA. Expression of EMT markers was analysed by qPCR and western blot. Intestinal fibrosis was induced in vivo using the heterotopic transplant model in WT and Sucnr1−/− mice and expression of EMT markers was analysed by qPCR and by confocal microscopy. Statistical analysis was performed with one-way ANOVA followed by Newman–Keuls test. Correlations were analysed with the Spearman’s coefficient. Results In intestinal resections from B3-CD patients, SUCNR1 mRNA expression was significantly increased when compared with B2-CD or non-IBD controls and it correlates positively with the mRNA expression of Snail1 and Snail2 and negatively with that of E-Cadherin. In the fistula tract, SUCNR1 is expressed in intestinal epithelial cells and lamina propria cells of the submucosa with a higher intensity in cells close to the fistula tract than in more distant areas. Succinate induced, in a dose–response manner, a significant increase in Vimentin, Snail1 and Snail2 expression and a reduction in E-cadherin expression. This effect was completely abolished when SUCNR1 was transiently knocked-down. WT-grafts 7 days after surgery exhibited: (a) an increase in gene and protein expression of SUCNR1, (b) an increase in the expression of Vimentin, Snail1 and Snail2 and a significant reduction in E-Cadherin compared with WT-grafts at day 0. Of interest, KO-grafts at day 7 failed to exhibited changes in the expression of Vimentin, Snail1, Snail2 or E-Cadherin. Conclusion SUCNR1 is increased specifically in the fistula tract of B3-CD patients. It mediates EMT in intestinal epithelial cells and in a murine model of fibrosis. Hence, SUCNR1 might be a potential pharmacological target for fistula treatment. Reference

PM2.5 Upregulates the Expression of MUC5AC via the EGFR-PI3K Pathway in Human Sinonasal Epithelial Cells

2021 ◽  
pp. 1-14
Author(s):  
Jian Jiao ◽  
Puqi Hu ◽  
Ying Li ◽  
Chao Cai ◽  
Xiangdong Wang ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epithelial Cells ◽  
Protein Expression ◽  
Mrna Expression ◽  
Pi3k Pathway ◽  
Cell Cycle Protein ◽  
Human Nasal Epithelial Cells ◽  
Mrna And Protein Expression ◽  
Pm2.5 Exposure ◽  
Β Tubulin

<b><i>Background:</i></b> Fine particulate matter (PM) (PM with an aerodynamic diameter &#x3c;2.5 μm, PM2.5) exposure contributes to respiratory disease development and exacerbation. <b><i>Objective:</i></b> We sought to investigate the effect of PM2.5 exposure on mucociliary function in primary human nasal epithelial cells (HNECs) and the underlying mechanism. <b><i>Methods:</i></b> HNECs derived from control subjects and patients with chronic rhinosinusitis with nasal polyps were established as air-liquid interface cultures. Confluent cultures were exposed to 100 or 200 μg/mL PM2.5 for 24 h and assessed for expression of specific mucociliary-associated factors, the percentage of β-tubulin IV-positive and MUC5AC-positive cells, expression of epidermal growth factor receptor (EGFR) ligand and activation of phosphoinositide 3-kinase (PI3K)-AKT/ERK. In addition, cultures pretreated for 30 min with AG1478 (an EGFR inhibitor) or LY294002 (a PI3K inhibitor) following PM2.5 exposure were assessed for MUC5AC mRNA and protein expression. <b><i>Results:</i></b> PM2.5 exposure at 100 or 200 μg/mL for 24 h did not affect geminin coiled-coil domain containing, multiciliate differentiation and DNA synthesis associated cell cycle protein, FOXJ1, or DNAI2 mRNA expression or the percentage of β-tubulin IV-positive cells. However, 200 μg/mL PM2.5 exposure significantly increased mRNA expression of SAM-pointed domain-containing ETS transcription factor and MUC5AC and the percentage of MUC5AC-positive cells. PM2.5 also increased expression of EGFR ligands, including heparin-binding EGF-like growth factor and amphiregulin. Furthermore, PM2.5induced activation of PI3K, AKT, and ERK, and pretreatment of HNECs with AG1478 or LY294002 attenuated PM2.5-induced MUC5AC mRNA and protein expression. <b><i>Conclusions and Clinical Relevance:</i></b> This study demonstrates that short-term PM2.5 exposure increases MUC5AC expression in HNECs. Furthermore, this study shows that PM2.5-induced MUC5AC expression is likely mediated through the EGFR-PI3K pathway.

scholarly journals Dysregulation of protein kinase C in adult depression and suicide: evidence from postmortem brain studies

2021 ◽  
Author(s):  
Ghanshyam N Pandey ◽  
Anuradha Sharma ◽  
Hooriyah S Rizavi ◽  
Xinguo Ren
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Protein Expression ◽  
Mrna Expression ◽  
Postmortem Brain ◽  
Cortical Region ◽  
Cytosol Fraction ◽  
Kinase C ◽  
Pkc Isozymes

Abstract Background Several lines of evidence suggest the abnormalities of protein kinase C (PKC) signaling system in mood disorders and suicide based primarily on the studies of PKC and its isozymes in the platelets and postmortem brain of depressed and suicidal subjects. In this study we examined the role of PKC isozymes in depression and suicide. Methods We determined the protein and mRNA expression of various PKC isozymes in the prefrontal cortical region [Brodmann area 9 (BA9)] in 24 normal control (NC) subjects, 24 depressed suicide (DS) subjects and 12 depressed non-suicide (DNS) subjects. The levels of mRNA in the prefrontal cortex (PFC) were determined by qRT-PCR and the protein expression was determined by Western blotting. Results We observed a significant decrease in mRNA expression of PKCα, PKCβI, PKCδ and PKCε and decreased protein expression either in the membrane or the cytosol fraction of PKC isozymes - PKCα, PKCβI, PKCβII and PKCδ in DS and DNS subjects compared with NC subjects. Conclusions The current study provides detailed evidence of specific dysregulation of certain PKC isozymes in the postmortem brain of DS and DNS subjects and further supports earlier evidence for the role of PKC in the platelets and brain of adult and teenage depressed and suicidal population. This comprehensive study may lead to further knowledge of the involvement of PKC in the pathophysiology of depression and suicide.

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