scholarly journals Influence of high glucose in the expression of miRNAs and IGF1R signaling pathway in human myometrial explants

2021 ◽  
Author(s):  
Rodolfo R. Favaro ◽  
Diana M. Morales-Prieto ◽  
Jörg Herrmann ◽  
Jürgen Sonnemann ◽  
Ekkehard Schleussner ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Signaling Pathway ◽  
High Glucose ◽  
Human Myometrium ◽  
Mrna Levels ◽  
Group Data ◽  
Short Term Exposure ◽  
Low Glucose ◽  
Biochemical Analyses ◽  
Igf1r Signaling

Abstract Purpose Several roles are attributed to the myometrium including sperm and embryo transport, menstrual discharge, control of uterine blood flow, and labor. Although being a target of diabetes complications, the influence of high glucose on this compartment has been poorly investigated. Both miRNAs and IGF1R are associated with diabetic complications in different tissues. Herein, we examined the effects of high glucose on the expression of miRNAs and IGF1R signaling pathway in the human myometrium. Methods Human myometrial explants were cultivated for 48 h under either high or low glucose conditions. Thereafter, the conditioned medium was collected for biochemical analyses and the myometrial samples were processed for histological examination as well as miRNA and mRNA expression profiling by qPCR. Results Myometrial structure and morphology were well preserved after 48 h of cultivation in both high and low glucose conditions. Levels of lactate, creatinine, LDH and estrogen in the supernatant were similar between groups. An explorative screening by qPCR arrays revealed that 6 out of 754 investigated miRNAs were differentially expressed in the high glucose group. Data validation by single qPCR assays confirmed diminished expression of miR-215-5p and miR-296-5p, and also revealed reduced miR-497-3p levels. Accordingly, mRNA levels of IGF1R and its downstream mediators FOXO3 and PDCD4, which are potentially targeted by miR-497-3p, were elevated under high glucose conditions. In contrast, mRNA expression of IGF1, PTEN, and GLUT1 was unchanged. Conclusions The human myometrium responds to short-term exposure (48 h) to high glucose concentrations by regulating the expression of miRNAs, IGF1R and its downstream targets.

scholarly journals The Onset of Labor Alters Corticotropin-Releasing Hormone Type 1 Receptor Variant Expression in Human Myometrium: Putative Role of Interleukin-1β

Endocrinology ◽  
2007 ◽  
Vol 148 (7) ◽  
pp. 3205-3213 ◽  
Author(s):  
Danijela Markovic ◽  
Manu Vatish ◽  
Mei Gu ◽  
Donna Slater ◽  
Rob Newton ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Nuclear Translocation ◽  
Fold Increase ◽  
Nuclear Factor Κb ◽  
Prolonged Treatment ◽  
Human Myometrium ◽  
Mrna Levels ◽  
Myometrial Cells ◽  
Important Regulator ◽  
R1 Gene

CRH targets the human myometrium during pregnancy. The efficiency of CRH actions is determined by expression of functional receptors (CRH-R), which are dynamically regulated. Studies in myometrial tissue biopsies using quantitative RT-PCR demonstrated that the onset of labor, term or preterm, is associated with a significant 2- to 3-fold increase in CRH-R1 mRNA levels. Detailed analysis of myometrial CRH-R1 mRNA variants showed a decline of the pro-CRH-R1 mRNA encoding the CRH-R1β variant during labor and increased mRNA levels of CRH-R1d mRNA. Studies in myometrial cells identified IL-1β as an important regulator of myometrial CRH-R1 gene expression because prolonged treatment of myometrial cells with IL-1β (1 ng/ml) for 18 h induced expression of CRH-R1 mRNA levels by 1.5- to 2-fold but significantly attenuated CRH-R1β mRNA expression by 70%. In contrast, IL-1β had no effect on CRH-R1d mRNA expression. Studies using specific inhibitors suggest that ERK1/2, p38 MAPK, and downstream nuclear translocation of nuclear factor-κB mediate IL-1β effects on myometrial CRH-R1 gene. However, the increased CRH-R1 mRNA expression was associated with a dampening of the receptor efficacy to activate the adenylyl cyclase/cAMP signaling cascade. Thus, our findings suggest that IL-1β is an important regulator of CRH-R1 expression and functional activity, and this interaction might play a role in the transition of the uterus from quiescence to active contractions necessary for the onset of parturition.

scholarly journals Molecular cloning of a novel secreted peptide, INM02, and regulation of its expression by glucose

2009 ◽  
Vol 202 (3) ◽  
pp. 355-364 ◽  
Author(s):  
Xuanchun Wang ◽  
Wei Gong ◽  
Yu Liu ◽  
Zhihong Yang ◽  
Wenbai Zhou ◽  
...  
Keyword(s):  
Pancreatic Islets ◽  
High Glucose ◽  
Northern Blot Analysis ◽  
Polyclonal Antibodies ◽  
Rat Tissues ◽  
Mrna Levels ◽  
Bladder Tissue ◽  
Reading Frame ◽  
Low Glucose

We report the identification of a novel secreted peptide, INM02. The mRNA transcript of human INM02 gene is about 3.0 kb. Its open-reading frame contains 762 bps and encodes a protein of 254 amino acids. Northern blot analysis demonstrates that INM02 mRNA is widely expressed in rat tissues, especially with abundant quantities in pancreatic islets, testis, and bladder tissue. We have expressed recombinant INM02 protein and generated rabbit anti-INM02 polyclonal antibodies. We show here that INM02 could be detectable in human serum by ELISA. We also present evidence that INM02 mRNA expression could be regulated by glucose. Experiments on both MIN6 cells and intact isolated islets demonstrate that INM02 mRNA levels are increased more than threefold by high glucose (25 mM) when compared with low glucose (5.5 mM). ELISA analysis shows that secretion of INM02 is significantly augmented by high glucose in vitro. It is speculated that as a novel secreted protein, INM02 is associated with functions of pancreatic islets, especially of β-cells.

scholarly journals The Role of Cyclic-ADP-Ribose-Signaling Pathway in Oxytocin-Induced Ca2+ Transients in Human Myometrium Cells

Endocrinology ◽  
2004 ◽  
Vol 145 (2) ◽  
pp. 881-889 ◽  
Author(s):  
Hosana Barata ◽  
Michael Thompson ◽  
Weronika Zielinska ◽  
Young S. Han ◽  
Carlos B. Mantilla ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
Uterine Contraction ◽  
Cyclase Activity ◽  
Human Myometrium ◽  
Mrna Levels ◽  
Myometrial Cells ◽  
Cd38 Expression ◽  
Cyclic Adp Ribose ◽  
Multiple Signaling

Abstract Human myometrial contraction plays a fundamental role in labor. Dysfunction of uterine contraction is an important cause of labor progression failure. Although the mechanisms controlling uterine contraction are not completely understood, intracellular Ca2+ mobilization plays an important role during uterine contraction. Several mechanisms of intracellular Ca2+ mobilization are present in smooth muscle, but in the human uterus, only 1,4,5-trisphosphate-induced Ca2+ release has been studied extensively. Ryanodine receptor channels are present in myometrium. We determined the role of the cyclic ADP-ribose (cADPR)-signaling pathway in oxytocin-induced intracellular Ca2+ [(Ca2+)i] transients in human myometrial cells. We found that oxytocin-induced Ca2+ transient is dependent on several sources of Ca2+, including extracellular Ca2+ and intracellular Ca2+ stores. In addition, we found that both the 1,4,5-trisphosphate- and the cADPR-induced Ca2+ releasing systems are important for the induction of [Ca2+]i transients by oxytocin in human myometrial cells. Furthermore, we investigated TNFα regulation of oxytocin-induced [Ca2+]i transients, CD38 cyclase activity, and CD38 expression in human myometrial cells. We found that oxytocin-induced [Ca2+]i transients were significantly increased by 50 ng/ml TNF. Similarly, CD38 mRNA levels, CD38 expression, and cyclase activity were increased by TNFα, thus increasing cADPR levels. We propose that a complex interaction between multiple signaling pathways is important for the development of intracellular Ca2+ transients induced by oxytocin and that TNFα may contribute for the myometrium preparation for labor by regulating the cADPR-signaling pathway. The observation that the cADPR-signaling pathway is important for the development of intracellular Ca2+ transients in human myometrial cells raises the possibility that this signaling pathway could serve as a target for the development of new therapeutic strategies for abnormal myometrial contraction observed during pregnancy.

scholarly journals Resveratrol attenuates diabetes-associated cell centrosome amplification via inhibiting the PKCα-p38 to c-myc/c-jun pathway

2019 ◽  
Vol 52 (1) ◽  
pp. 72-83 ◽  
Author(s):  
Qigui Wu ◽  
Xiaoyu Chen ◽  
Qinju He ◽  
Lang Lang ◽  
Peng Xu ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Palmitic Acid ◽  
Signaling Pathway ◽  
High Glucose ◽  
Mononuclear Cells ◽  
Centrosome Amplification ◽  
Mrna Levels ◽  
Peripheral Blood Mononuclear ◽  
Protein Levels

Abstract Type 2 diabetes increases the risk for cancer. Centrosome amplification can initiate tumorigenesis. We have described that type 2 diabetes increases the centrosome amplification of peripheral blood mononuclear cells, with high glucose, insulin, and palmitic acid as the triggers, which suggests that centrosome amplification is a candidate biological mechanism linking diabetes to cancer. In this study, we aimed to further investigate the signaling pathways of the diabetes-associated centrosome amplification and to examine whether and how resveratrol inhibits the centrosome amplification. The results showed that treatment with high glucose, insulin, and palmitic acid, alone or in combination, could increase the protein levels of phospho-protein kinase C alpha (p-PKCα), phospho-p38 mitogen-activated protein kinases (p-p38), c-myc, and c-jun, as well as the mRNA levels of c-myc and c-jun. PKCα inhibitor could inhibit the treatment-induced increase in the protein levels of p-p38, c-myc, and c-jun. Inhibitor or siRNA of p38 was also able to inhibit the treatment-induced increase in the levels of p-p38, c-myc, and c-jun. Meanwhile, knockdown of c-myc or c-jun did not alter the treatment-induced increase in the phosphorylation of PKCα or p38. Importantly, inhibition of the phosphorylation of PKCα or p38 and knockdown of c-myc or c-jun could attenuate the centrosome amplification. In diabetic mice, the levels of p-PKCα, p-p38, c-myc, and c-jun were all increased in the colon tissues. Interestingly, resveratrol, but not metformin, was able to attenuate the treatment-induced increase in the levels of p-PKCα, p-p38, c-myc, and c-jun, as well as the centrosome amplification. In conclusion, our results suggest that PKCα-p38 to c-myc/c-jun is the signaling pathway of the diabetes-associated centrosome amplification, and resveratrol attenuates the centrosome amplification by inhibiting this signaling pathway.

Modulation of IGF1R Signaling Pathway by GIGYF1 in High Glucose-Induced SHSY-5Y Cells

2018 ◽  
Vol 37 (12) ◽  
pp. 1044-1054
Author(s):  
Ting Zhou ◽  
Yuefei Ma ◽  
Juan Tang ◽  
Fengqi Guo ◽  
Mingxia Dong ◽  
...  
Keyword(s):  
Signaling Pathway ◽  
High Glucose ◽  
Igf1r Signaling

scholarly journals KISS1 Suppresses Apoptosis and Stimulates the Synthesis of E2 in Porcine Ovarian Granulosa Cells

Animals ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 54 ◽  
Author(s):  
Xiaoping Xin ◽  
Zhonghui Li ◽  
Yuyi Zhong ◽  
Qingqing Li ◽  
Jiaying Wang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Mrna Expression ◽  
Signaling Pathway ◽  
Granulosa Cells ◽  
Cell Apoptosis ◽  
Pi3k Signaling ◽  
Mrna Levels ◽  
Ovarian Granulosa Cells ◽  
Estrogen Synthesis ◽  
Pi3k Signaling Pathway

Previous studies have strongly recommended that KISS-1 metastasis suppressor (KISS1) plays an essential gatekeeper of the initiation of reproductive maturation in mammals. However, KISS1 has been recently reported to highly express in ovarian granulosa cells (GCs). But the biological functionalities of KISS1 on cell apoptosis, cell cycle, and synthesis of estradiol-17β (E2) have not been explored in GCs. In this study, using porcine GCs as a cellular model, the overexpression plasmid of KISS1 was built to explore the biological effects of KISS1 on the PI3K signaling pathway, estrogen signaling pathway, cell apoptosis, cell cycle, and E2 secretion. We found that mRNA of KISS1 highly expressed in the ovary and significantly increased from immature to mature follicles in gilts. Overexpression of KISS1 could significantly increase the mRNA expression of PIK3CG, PIK3C1, and PDK1, and significantly decreased the mRNA levels of FOXO3, TSC2, and BAD of PI3K signaling pathway. Furthermore, results of the flow cytometry showed that overexpression of KISS1 significantly inhibited the apoptosis of GCs and decreased the percentage of GCs at G0/G1 phase of the cell cycle. Additionally, overexpression of KISS1 could increase the mRNA levels of Star, CYP17, 3B-HSD, 17B-HSD of estrogen synthesis signaling pathway, significantly increase the concentration of E2 in the supernatant of the cultured GCs, and up-regulate the mRNA expression levels of ESR1 and ESR2. These results suggested that KISS1 might suppress cell apoptosis through activating the PI3K signaling pathway and stimulate synthesis of E2 via boosting the estrogen synthesis signaling pathway. This study would be of great interests for exploring the biological functionalities of KISS1 in the folliculogenesis and sex steroid production of the ovaries in mammals.

scholarly journals Resveratrol inhibits high glucose-induced activation and cytokine production of isolated primary pancreatic stellate cells

2019 ◽  
Vol 8 (3) ◽  
pp. 35-47
Author(s):  
Zhen Zhou ◽  
Xiaodong Sun ◽  
Rao Yan ◽  
Jinfeng An ◽  
Xinjian Zhou ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
High Glucose ◽  
Stellate Cells ◽  
Collagen I ◽  
Pancreatic Stellate Cells ◽  
Mrna Levels ◽  
Tnf Α ◽  
Low Glucose ◽  
Glucose Treatment ◽  
Pharmacologically Active

Objective: Activation of pancreatic stellate cells (PSCs) is detrimental to pancreas function by promoting pancreatic fibrosis. Resveratrol is a natural and pharmacologically active compound. This study is to investigate the effect of resveratrol on the bilogical behavior of PSCs under high glucose condition.Methods: Isolated primary mouse PSCs were cultured in low glucose ( 5.5 mmol/L glucose, LG group ) medium, high glucose ( 25 mmol/L glucose, HG group ) medium and treated with  resveratrol ( 25 μmol/L or 50 μmol/L). Cell proliferation was examined using MTT assay. The expression of α-SMA and collagen I were determined using Western blotting. Alpha-SMA expression was also determined using immunocytochemistry staining. IL-1, IL-6, and TNF-α mRNA levels and secretion levels in media of PSCs were determined using qRT-PCR and ELISA respectively.Results: Cell Proliferation,  α-SMA and collagen I  expression levels, IL-1, IL-6, and TNF-α mRNA levels and secretion levels of PSCs were increased after high glucose treatment, compared with low glucose treatment. They were significantly decreased in PSCs treated with both high glucose and resveratrol, compared with high glucose treatment.Conclusion: Resveratrol inhibited high glucose induced PSCs proliferation, activation，cytokine expression and secretion in PSCs. Therefore, resveratrol can be potentially used in therapy of diseases such as type 2 diabetes mellitus (T2DM), pancreatitis and pancreatic cancer where PSCs is activated by high glucose.

scholarly journals Mechanical strain- and high glucose-induced alterations in mesangial cell collagen metabolism: role of TGF-beta.

1998 ◽  
Vol 9 (5) ◽  
pp. 827-836
Author(s):  
B L Riser ◽  
P Cortes ◽  
J Yee ◽  
A K Sharba ◽  
K Asano ◽  
...  
Keyword(s):  
High Glucose ◽  
Collagen Synthesis ◽  
Transforming Growth Factor ◽  
Mechanical Strain ◽  
Collagen Metabolism ◽  
Mrna Levels ◽  
Glucose Medium ◽  
Tgf Beta ◽  
Collagen Accumulation ◽  
Low Glucose

Cultured mesangial cells (MC) exposed to cyclic mechanical strain or high glucose levels increase their secretion of transforming growth factor-beta1 (TGF-beta1) and collagen, suggesting possible mechanisms for the development of diabetic renal sclerosis resulting from intraglomerular hypertension and/or hyperglycemia. This study examines whether glucose interacts with mechanical strain to influence collagen metabolism and whether this change is mediated by TGF-beta. Accordingly, rat MC were grown on flexible-bottom plates in 8 or 35 mM glucose media, subjected to 2 to 5 d of cyclic stretching, and assayed for TGF-beta1 mRNA, TGF-beta1 secretion, and the incorporation of 14C-proline into free or protein-associated hydroxyproline to assess the dynamics of collagen metabolism. Stretching or high glucose exposure increased TGF-beta1 secretion twofold and TGF-beta1 mRNA levels by 30 and 45%, respectively. However, the combination of these stimuli increased secretion greater than fivefold without further elevating mRNA. In 8 mM glucose medium, stretching significantly increased MC collagen synthesis and breakdown, but did not alter accumulation, whereas those stretched in 35 mM glucose markedly increased collagen accumulation. TGF-beta neutralization significantly reduced baseline collagen synthesis, breakdown, and accumulation in low glucose, but had no significant effect on the changes induced by stretch. In contrast, the same treatment of MC in high glucose medium greatly reduced stretch-induced synthesis and breakdown of collagen and totally abolished the increase in collagen accumulation. These results indicate that TGF-beta plays a positive regulatory role in MC collagen synthesis, breakdown, and accumulation. However, in low glucose there is no stretch-induced collagen accumulation, and the effect of TGF-beta is limited to basal collagen turnover. In high glucose media, TGF-beta is a critical mediator of stretch-induced collagen synthesis and catabolism, and, most importantly, its net accumulation. These data have important implications for the pathogenesis and treatment of diabetic glomerulosclerosis.

scholarly journals VDR Activation Reduces Proteinuria and High-Glucose-Induced Injury of Kidneys and Podocytes by Regulating Wnt Signaling Pathway

2017 ◽  
Vol 43 (1) ◽  
pp. 39-51 ◽  
Author(s):  
Jia Guo ◽  
Congqun Lu ◽  
Fangxing Zhang ◽  
Haixia Yu ◽  
Mengwen Zhou ◽  
...  
Keyword(s):  
Wnt Signaling ◽  
Signaling Pathway ◽  
High Glucose ◽  
Kidney Diseases ◽  
Wnt Signaling Pathway ◽  
Clinical Manifestations ◽  
Mrna Levels ◽  
Gsk 3Β ◽  
Stage Renal Disease ◽  
End Stage

Background: Diabetic nephropathy (DN) is a major cause of end-stage renal disease and proteinuria is one of the most prominent clinical manifestations. The expression of Vitamin D receptor (VDR) in patients with chronic kidney diseases was decreased, while VDR agonists could partially alleviate the proteinuria of DN in animal models. The present study was designed to determine the expression of VDR in renal tissues and its relationship with proteinuria the diabetic model db/db mice. Methods: The regulation effects of VDR on the Wnt signaling pathway were analyzed using RNA interference and VDR agonist paricalcitol. Results: With the increase in age of the db/db mice, the VDR protein and mRNA levels in renal tissues were decreased, proteinuria increased, and the protein and mRNA levels of GSK-3β of and β-catenin increased. Paricalcitol treatment resulted in the up-regulation of VDR and down-regulation of GSK-3β and β-catenin, indicating that VDR had a regulatory effect on the Wnt signaling pathway. Conclusion: VDR activation could reduce proteinuria of DN mice and alleviate high-glucose-induced injury of kidneys and podocytes by regulating the key molecules of Wnt signaling pathway.

scholarly journals Improvement of high-glucose and insulin resistance of chromium malate in 3T3-L1 adipocytes by glucose uptake and insulin sensitivity signaling pathways and its mechanism

RSC Advances ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 114-127 ◽  
Author(s):  
Weiwei Feng ◽  
Yongchao Liu ◽  
Fan Fei ◽  
Yao Chen ◽  
Yangyang Ding ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Insulin Sensitivity ◽  
Mrna Expression ◽  
Glucose Uptake ◽  
High Glucose ◽  
Mrna Levels ◽  
Related Protein ◽  
Model Group ◽  
Insulin Resistant ◽  
Chromium Malate

Chromium malate could increase the related protein and mRNA levels in 3T3-L1 adipocytes with insulin resistant. Pretreatment with the inhibitor completely/partially inhibited the GLUT-4 and Irs-1 proteins and mRNA expression compared to model group.

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