Glucose Promotes the Production of Interleukine-1β and Cyclooxygenase-2 in Mesangial Cells via Enhanced (Pro)Renin Receptor Expression

Abstract (Pro)renin receptor (PRR) is present in renal glomeruli, and its expression is up-regulated in diabetes. Similarly, renal inflammation is increased in the presence of hyperglycemia. The linkage between PRR and renal inflammation is not well established. We hypothesized that glucose-induced up-regulation of PRR leads to increased production of the proinflammatory factors IL-1β and cyclooxygenase-2 (COX-2). Studies were conducted in rat mesangial cells (RMCs) exposed to 30 mmd-glucose for 2 wk followed by PRR small interfering RNA knockdown, IL-1 receptor blockade with IL-1 receptor antagonist or angiotensin II type 1 receptor blockade with valsartan. The results showed that d-glucose treatment up-regulates prorenin, renin, angiotensin II, PRR, IL-1β, and COX-2 mRNA and protein expression and increases phosphorylation of ERK1/2, c-Jun N-terminal kinase, c-Jun, and nuclear factor-κB (NF-κB) p65 (serine 276,468 and 536), respectively. PRR small interfering RNA attenuated PRR, IL-1β, and COX-2 mRNA and protein expressions and significantly decreased angiotensin II production and phosphorylation of ERK1/2 and NF-κB p65 associated with high glucose exposure. Similarly, IL-1 receptor antagonist significantly reduced COX-2 mRNA and protein expression induced by high glucose. COX-2 inhibition reduced high-glucose-induced PRR expression. We conclude that glucose induces the up-regulation of PRR and its ligands prorenin and renin, leading to increased IL-1β and COX-2 production via the angiotensin II-dependent pathway. It is also possible that PRR could enhance the production of these inflammatory cytokines through direct stimulation of ERK1/2-NF-κB signaling cascade.

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Metformin Regulating miR-34a Pathway to Inhibit Egr1 in Rat Mesangial Cells Cultured with High Glucose

International Journal of Endocrinology ◽

10.1155/2018/6462793 ◽

2018 ◽

Vol 2018 ◽

pp. 1-15 ◽

Cited By ~ 7

Author(s):

Can Wu ◽

Ningning Qin ◽

Huiwen Ren ◽

Min Yang ◽

Shuang Liu ◽

...

Keyword(s):

Western Blot ◽

Inflammatory Cytokines ◽

High Glucose ◽

Real Time Pcr ◽

Small Interfering Rna ◽

Mesangial Cells ◽

Inflammatory Factors ◽

Quantitative Real Time Pcr ◽

Interfering Rna ◽

Cells Cultured

Background. Activating AMPKα negatively regulates Egr1 to inhibit inflammatory cytokines in high glucose. miR-34a inhibition increases phosphorylated AMPKα through mediating SIRT1 to suppress the development of fatty liver. Aim of the Study. To clarify the function of Egr1 on the inflammation and fibrosis in high glucose-cultured MCs, as well as to explore the effects of metformin on miR-34a pathway and Egr1 expression. Methods. We transfected MCs with miR-34a inhibitor. And MCs were transfected with small interfering RNA for silencing Egr1 and SIRT1. Quantitative real-time PCR was used to assay the transcription levels of Egr1 mRNA and miR-34a. Western blot was used to test the protein. And ELISA was used to measure inflammatory factors. Results. High glucose upregulates Egr1 to aggravate the inflammation and fibrosis in MCs. miR-34a suppresses the activation of SIRT1/AMPKα and results in promoting Egr1 in high glucose-cultured MCs. Metformin attenuates high glucose-stimulated inflammation and fibrosis in MCs by regulating miR-34a-mediated SIRT1/AMPKα activity and the downstream Egr1 protein. Conclusion. We enriched the effects of miR-34a pathway regulating Egr1 in high glucose-cultured MCs. It provides a foundation for future researches considering Egr1 as a therapeutic target and a new direction for the clinical application of metformin in early DKD.

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Impairment of hepatic nuclear factor-4α binding to the Stim1 promoter contributes to high glucose-induced upregulation of STIM1 expression in glomerular mesangial cells

AJP Renal Physiology ◽

10.1152/ajprenal.00563.2014 ◽

2015 ◽

Vol 308 (10) ◽

pp. F1135-F1145 ◽

Cited By ~ 6

Author(s):

Yanxia Wang ◽

Sarika Chaudhari ◽

Yuezhong Ren ◽

Rong Ma

Keyword(s):

Protein Expression ◽

High Glucose ◽

Mesangial Cells ◽

Therapeutic Option ◽

Binding Activity ◽

Nuclear Factor ◽

Glomerular Mesangial Cells ◽

Expression Levels ◽

293 Cells ◽

Interfering Rna

The present study was carried out to investigate if hepatic nuclear factor (HNF)4α contributed to the high glucose-induced increase in stromal interacting molecule (STIM)1 protein abundance in glomerular mesangial cells (MCs). Western blot and immunofluorescence experiments showed HNF4α expression in MCs. Knockdown of HNF4α using a small interfering RNA approach significantly increased mRNA expression levels of both STIM1 and Orai1 and protein expression levels of STIM1 in cultured human MCs. Consistently, overexpression of HNF4α reduced expressed STIM1 protein expression in human embryonic kidney-293 cells. Furthermore, high glucose treatment did not significantly change the abundance of HNF4α protein in MCs but significantly attenuated HNF4α binding activity to the Stim1 promoter. Moreover, knockdown of HNF4α significantly augmented store-operated Ca2+ entry, which is known to be gated by STIM1 and has recently been found to be antifibrotic in MCs. In agreement with those results, knockdown of HNF4α significantly attenuated the fibrotic response of high glucose. These results suggest that HNF4α negatively regulates STIM1 transcription in MCs. High glucose increases STIM1 expression levels by impairing HNF4α binding activity to the Stim1 promoter, which subsequently releases Stim1 transcription from HNF4α repression. Since the STIM1-gated store-operated Ca2+ entry pathway in MCs has an antifibrotic effect, inhibition of HNF4α in MCs might be a potential therapeutic option for diabetic kidney disease.

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Glibenclamide Induces Collagen IV Catabolism in High Glucose-Stimulated Mesangial Cells

Experimental Diabetes Research ◽

10.1155/2012/183535 ◽

2012 ◽

Vol 2012 ◽

pp. 1-9 ◽

Cited By ~ 2

Author(s):

Liping Zhu ◽

Pedro Cortes ◽

Clare Hassett ◽

David W. Taube ◽

Jerry Yee

Keyword(s):

Protein Expression ◽

High Glucose ◽

Mesangial Cells ◽

Collagen Iv ◽

Diabetic Rats ◽

Protein Accumulation ◽

Generating Capacity ◽

Mrna And Protein Expression ◽

Low Dosage ◽

Pai 1

We have shown the full prevention of mesangial expansion in insulin-deficient diabetic rats by treatment with clinically-relevant dosages of glibenclamide (Glib). Studies in mesangial cells (MCs) also demonstrated reduction in the high glucose (HG)-induced accumulation of collagens, proposing that this was due to increased catabolism. In the present study, we investigated the signaling pathways that may be implicated in Glib action. Rat primary MCs were exposed to HG for 8 weeks with or without Glib in therapeutic (0.01 μM) or supratherapeutic (1.0 μM) concentrations. We found that HG increased collagen IV protein accumulation and PAI-1 mRNA and protein expression, in association with decreased cAMP generating capacity and decreased PKA activity. Low Glib increased collagen IV mRNA but fully prevented collagen IV protein accumulation and PAI-1 overexpression while enhancing cAMP formation and PKA activity. MMP2 mRNA, protein expression and gelatinolytic activity were also enhanced. High Glib was, overall, ineffective. In conclusion, low dosage/concentration Glib prevents HG-induced collagen accumulation in MC by enhancing collagen catabolism in a cAMP-PKA-mediated PAI-1 inhibition.

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Aldosterone induces interleukin-18 through endothelin-1, angiotensin II, Rho/Rho-kinase, and PPARs in cardiomyocytes

AJP Heart and Circulatory Physiology ◽

10.1152/ajpheart.00148.2008 ◽

2008 ◽

Vol 295 (3) ◽

pp. H1279-H1287 ◽

Cited By ~ 31

Author(s):

Takashi Doi ◽

Tsuyoshi Sakoda ◽

Takafumi Akagami ◽

Toshio Naka ◽

Yoshitomo Mori ◽

...

Keyword(s):

Angiotensin Ii ◽

Protein Expression ◽

Receptor Antagonist ◽

Myocardial Dysfunction ◽

Rho Kinase ◽

Ang Ii ◽

Rat Cardiomyocytes ◽

Endothelin 1 ◽

Ppar Agonists ◽

Mrna And Protein Expression

Aldosterone (Aldo) is recognized as an important risk factor for cardiovascular diseases. IL-18 induces myocardial hypertrophy, loss of contractility of cardiomyocytes, and apoptosis leading myocardial dysfunction. However, so far, there have been few reports concerning the interaction between Aldo and IL-18. The present study examined the effects and mechanisms of Aldo on IL-18 expression and the roles of peroxisome proliferator-activated receptor (PPAR) agonists in rat cardiomyocytes. We used cultured rat neonatal cardiomyocytes stimulated with Aldo to measure IL-18 mRNA and protein expression, Rho-kinase, and NF-κB activity. We also investigated the effects of PPAR agonists on these actions. Aldo, endothelin-1 (ET-1), and angiotensin II (ANG II) increased IL-18 mRNA and protein expression. Mineralocorticoid receptor antagonists, endothelin A receptor antagonist, and ANG II receptor antagonist inhibited Aldo-induced IL-18 expression. Aldo induced ET-1 and ANG II production in cultured media. Moreover, Rho/Rho-kinase inhibitor and statin inhibited Aldo-induced IL-18 expression. On the other hand, Aldo upregulated the activities of Rho-kinase and NF-κB. PPAR agonists attenuated the Aldo-induced IL-18 expression and NF-κB activity but not the Rho-kinase activity. Our findings indicate that Aldo induces IL-18 expression through a mechanism that involves, at a minimum, ET-1 and ANG II acting via the Rho/Rho-kinase and PPAR/NF-κB pathway. The induction of IL-18 in cardiomyocytes by Aldo, ET-1, and ANG II might, therefore, cause a deterioration of the cardiac function in an autocrine and paracrine fashion. The inhibition of the IL-18 expression by PPAR agonists might be one of the mechanisms whereby the beneficial cardiovascular effects are exerted.

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Hypoxia increases KIAA1199/CEMIP expression and enhances cell migration in pancreatic cancer

Scientific Reports ◽

10.1038/s41598-021-97752-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Takuya Oba ◽

Norihiro Sato ◽

Yasuhiro Adachi ◽

Takao Amaike ◽

Yuzan Kudo ◽

...

Keyword(s):

Cell Migration ◽

Protein Expression ◽

Hypoxia Inducible Factor ◽

Ductal Adenocarcinoma ◽

Rt Pcr ◽

Transwell Migration Assay ◽

Migration Assay ◽

Hypoxic Conditions ◽

Mrna And Protein Expression ◽

Interfering Rna

AbstractPancreatic ductal adenocarcinoma (PDAC) is characterised by dense desmoplasia and hypoxic microenvironment. Our previous reports demonstrated that hyaluronan (HA), especially low-molecular-weight HA, provides a favourable microenvironment for PDAC progression. However, the effect of hypoxia on HA metabolism remains unknown. Using quantitative real-time RT-PCR and western blot analysis, we analysed the changes in the expression of HA-synthesizing enzymes (HAS2 and HAS3) and HA-degrading enzymes (HYAL1, KIAA1199/CEMIP) in PDAC cell lines under hypoxic conditions. Hypoxia increased the mRNA and protein expression of KIAA1199, whereas it decreased HYAL1 expression. The expression of HAS3 was increased and HAS2 remained unchanged in response to hypoxia. The effect of KIAA1199 on hypoxia-induced cell migration was determined using a transwell migration assay and small-interfering RNA (siRNA). Hypoxia enhanced the migratory ability of PDAC cells, which was inhibited by KIAA1199 knockdown. We also used immunohistochemistry to analyse the protein expression of hypoxia inducible factor (HIF) 1α and KIAA1199 in PDAC tissues. There was a significant immunohistochemically positive correlation between KIAA1199 and HIF1α. These findings suggest that hypoxia-induced KIAA1199 expression may contribute to enhanced motility in PDAC.

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Kallikrein gene ‘knock-down’ by small interfering RNA transfection induces a profibrotic phenotype in rat mesangial cells

Journal of Hypertension ◽

10.1097/hjh.0b013e3282f0ca68 ◽

2008 ◽

Vol 26 (1) ◽

pp. 93-101 ◽

Cited By ~ 7

Author(s):

Izabella ZA Pawluczyk ◽

Eddie KC Tan ◽

David Lodwick ◽

Kevin PG Harris

Keyword(s):

Small Interfering Rna ◽

Mesangial Cells ◽

Knock Down ◽

Rna Transfection ◽

Interfering Rna

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Inhibitory Effect of Sambucus sieboldiana var. pendula (Nakai) Extract on the mRNA and Protein Expression of iNOS and COX-2 in Raw 264.7 Cells

Microbiology and Biotechnology Letters ◽

10.4014/mbl.1704.04002 ◽

2017 ◽

Vol 45 (2) ◽

pp. 178-183 ◽

Cited By ~ 1

Author(s):

jin-Young Lee ◽

Dan-Hee Yoo ◽

Jung-Woo Chae

Keyword(s):

Protein Expression ◽

Inhibitory Effect ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Mrna And Protein Expression ◽

Cox 2

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Protein Kinase C-ε Induces Caveolin-Dependent Internalization of Vascular Adenosine 5′-Triphosphate–Sensitive K + Channels

Hypertension ◽

10.1161/hypertensionaha.108.110817 ◽

2008 ◽

Vol 52 (3) ◽

pp. 499-506 ◽

Cited By ~ 32

Author(s):

Jundong Jiao ◽

Vivek Garg ◽

Baofeng Yang ◽

Terry S. Elton ◽

Keli Hu

Keyword(s):

Smooth Muscle ◽

Protein Kinase C ◽

Angiotensin Ii ◽

Protein Kinase ◽

Cell Surface ◽

Small Interfering Rna ◽

Caveolin 1 ◽

Kinase C ◽

Interfering Rna ◽

Pkc Activation

Vascular ATP-sensitive K + (K ATP ) channels are critical regulators of arterial tone and, thus, blood flow in response to local metabolic needs. They are important targets for clinically used drugs to treat hypertensive emergency and angina. It is known that protein kinase C (PKC) activation inhibits K ATP channels in vascular smooth muscles. However, the mechanism by which PKC inhibits the channel remains unknown. Here we report that caveolin-dependent internalization is involved in PKC-ε–mediated inhibition of vascular K ATP channels (Kir6.1 and SUR2B) by phorbol 12-myristate 13-acetate or angiotensin II in human embryonic kidney 293 cells and immortalized human saphenous vein vascular smooth muscle cells. We showed that Kir6.1 substantially overlapped with caveolin-1 at the cell surface. Cholesterol depletion with methyl-β-cyclodextrin significantly reduced, whereas overexpression of caveolin-1 largely enhanced, PKC-induced inhibition of Kir6.1/SUR2B currents. Importantly, we demonstrated that activation of PKC-ε caused internalization of K ATP channels, the effect that was blocked by depletion of cholesterol with methyl-β-cyclodextrin, expression of dominant-negative dynamin mutant K44E, or knockdown of caveolin-1 with small interfering RNA. Moreover, patch-clamp studies revealed that PKC-ε–mediated inhibition of the K ATP current induced by PMA or angiotensin II was reduced by a dynamin mutant, as well as small interfering RNA targeting caveolin-1. The reduction in the number of plasma membrane K ATP channels by PKC activation was further confirmed by cell surface biotinylation. These studies identify a novel mechanism by which the levels of vascular K ATP channels could be rapidly downregulated by internalization. This finding provides a novel mechanistic insight into how K ATP channels are regulated in vascular smooth muscle cells.

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