Negative regulation of Smad1 pathway and collagen IV expression by store-operated Ca2+ entry in glomerular mesangial cells
Copyright © 2017 the American Physiological Society..
Collagen IV (Col IV) is a major component of expanded glomerular extracellular matrix in diabetic nephropathy and Smad1 is a key molecule regulating Col IV expression in mesangial cells (MCs). The present study was conducted to determine if Smad1 pathway and Col IV protein abundance were regulated by store-operated Ca2+ entry (SOCE). In cultured human MCs, pharmacological inhibition of SOCE significantly increased the total amount of Smad1 protein. Activation of SOCE blunted high-glucose-increased Smad1 protein content. Treatment of human MCs with ANG II at 1 µM for 15 min, high glucose for 3 days, or TGF-β1 at 5 ng/ml for 30 min increased the level of phosphorylated Smad1. However, the phosphorylation of Smad1 by those stimuli was significantly attenuated by activation of SOCE. Knocking down Smad1 reduced, but expressing Smad1 increased, the amount of Col IV protein. Furthermore, activation of SOCE significantly attenuated high-glucose-induced Col IV protein production, and blockade of SOCE substantially increased the abundance of Col IV. To further verify those in vitro findings, we downregulated SOCE specifically in MCs in mice using small-interfering RNA (siRNA) against Orai1 (the channel protein mediating SOCE) delivered by the targeted nanoparticle delivery system. Immunohistochemical examinations showed that expression of both Smad1 and Col IV proteins was significantly greater in the glomeruli with positively transfected Orai1 siRNA compared with the glomeruli from the mice without Orai1 siRNA treatment. Taken together, our results indicate that SOCE negatively regulates the Smad1 signaling pathway and inhibits Col IV protein production in MCs.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2017 |
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| Erschienen: |
2017 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:312 |
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| Enthalten in: |
American journal of physiology. Renal physiology - 312(2017), 6 vom: 01. Juni, Seite F1090-F1100 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Wu, Peiwen [VerfasserIn] |
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| Links: |
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| Anmerkungen: |
Date Completed 08.08.2017 Date Revised 08.05.2019 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1152/ajprenal.00642.2016 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM269883924 |
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| 100 | 1 | |a Wu, Peiwen |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Negative regulation of Smad1 pathway and collagen IV expression by store-operated Ca2+ entry in glomerular mesangial cells |
| 264 | 1 | |c 2017 | |
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| 500 | |a published: Print-Electronic | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a Copyright © 2017 the American Physiological Society. | ||
| 520 | |a Collagen IV (Col IV) is a major component of expanded glomerular extracellular matrix in diabetic nephropathy and Smad1 is a key molecule regulating Col IV expression in mesangial cells (MCs). The present study was conducted to determine if Smad1 pathway and Col IV protein abundance were regulated by store-operated Ca2+ entry (SOCE). In cultured human MCs, pharmacological inhibition of SOCE significantly increased the total amount of Smad1 protein. Activation of SOCE blunted high-glucose-increased Smad1 protein content. Treatment of human MCs with ANG II at 1 µM for 15 min, high glucose for 3 days, or TGF-β1 at 5 ng/ml for 30 min increased the level of phosphorylated Smad1. However, the phosphorylation of Smad1 by those stimuli was significantly attenuated by activation of SOCE. Knocking down Smad1 reduced, but expressing Smad1 increased, the amount of Col IV protein. Furthermore, activation of SOCE significantly attenuated high-glucose-induced Col IV protein production, and blockade of SOCE substantially increased the abundance of Col IV. To further verify those in vitro findings, we downregulated SOCE specifically in MCs in mice using small-interfering RNA (siRNA) against Orai1 (the channel protein mediating SOCE) delivered by the targeted nanoparticle delivery system. Immunohistochemical examinations showed that expression of both Smad1 and Col IV proteins was significantly greater in the glomeruli with positively transfected Orai1 siRNA compared with the glomeruli from the mice without Orai1 siRNA treatment. Taken together, our results indicate that SOCE negatively regulates the Smad1 signaling pathway and inhibits Col IV protein production in MCs | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Research Support, N.I.H., Extramural | |
| 650 | 4 | |a Research Support, Non-U.S. Gov't | |
| 650 | 4 | |a extracellular matrix | |
| 650 | 4 | |a store-operated calcium entry | |
| 650 | 7 | |a Calcium Channel Blockers |2 NLM | |
| 650 | 7 | |a Collagen Type IV |2 NLM | |
| 650 | 7 | |a ORAI1 Protein |2 NLM | |
| 650 | 7 | |a Orai1 protein, mouse |2 NLM | |
| 650 | 7 | |a SMAD1 protein, human |2 NLM | |
| 650 | 7 | |a Smad1 Protein |2 NLM | |
| 650 | 7 | |a Smad1 protein, mouse |2 NLM | |
| 650 | 7 | |a TGFB1 protein, human |2 NLM | |
| 650 | 7 | |a Transforming Growth Factor beta1 |2 NLM | |
| 650 | 7 | |a Angiotensin II |2 NLM | |
| 650 | 7 | |a 11128-99-7 |2 NLM | |
| 650 | 7 | |a Glucose |2 NLM | |
| 650 | 7 | |a IY9XDZ35W2 |2 NLM | |
| 700 | 1 | |a Ren, Yuezhong |e verfasserin |4 aut | |
| 700 | 1 | |a Ma, Yuhong |e verfasserin |4 aut | |
| 700 | 1 | |a Wang, Yanxia |e verfasserin |4 aut | |
| 700 | 1 | |a Jiang, Hui |e verfasserin |4 aut | |
| 700 | 1 | |a Chaudhari, Sarika |e verfasserin |4 aut | |
| 700 | 1 | |a Davis, Mark E |e verfasserin |4 aut | |
| 700 | 1 | |a Zuckerman, Jonathan E |e verfasserin |4 aut | |
| 700 | 1 | |a Ma, Rong |e verfasserin |4 aut | |
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