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Negative regulation of Smad1 pathway and collagen IV expression by store-operated Ca^(2+) entry in glomerular mesangial cells

Wu, Peiwen and Ren, Yuezhong and Ma, Yuhong and Wang, Yanxia and Jiang, Hui and Chaudhari, Sarika and Davis, Mark E. and Zuckerman, Jonathan E. and Ma, Rong (2017) Negative regulation of Smad1 pathway and collagen IV expression by store-operated Ca^(2+) entry in glomerular mesangial cells. American Journal of Physiology - Renal Physiology, 312 (6). F1090-F1100. ISSN 1931-857X. doi:10.1152/ajprenal.00642.2016.

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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 Ca^(2+) 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.

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Davis, Mark E.0000-0001-8294-1477
Additional Information:© 2017 American Physiology Society. Submitted 2 December 2016; accepted in final form 9 March 2017. Published 4 June 2017. We thank the Harry S. Moss Heart Trust for supporting this study and Dr. Xia Lin at the Baylor College of Medicine for providing Flag-tagged Smad1 expression plasmid. This work was supported by National Institute of Diabetes and Digestive and Kidney Diseases Grant 5RO1-DK-079968-01 (to R. Ma), by a Grant-in-Aid from the American Heart Association Southwestern Affiliate (16GRNT27780043, to R. Ma), by the National Natural Science Foundation of China (81400805, to P. Wu), and by the Natural Science Foundation of Fujian Province (2016J01449, to P. Wu). Author Contributions: P.W. and R.M. conceived and designed research; P.W., Y.R., Y.M., Y.W., H.J., and S.C. performed experiments; P.W., Y.R., Y.M., and R.M. analyzed data; P.W., Y.R., and R.M. interpreted results of experiments; P.W., Y.R., and R.M. prepared figures; P.W. and Y.R. drafted manuscript; P.W., Y.R., Y.M., Y.W., H.J., S.C., and R.M. approved final version of manuscript; M.E.D., J.E.Z., and R.M. edited and revised manuscript. No conflicts of interest, financial or otherwise, are declared by the authors.
Funding AgencyGrant Number
Harry S. Moss Heart TrustUNSPECIFIED
American Heart Association16GRNT27780043
National Science Foundation of China81400805
Natural Science Foundation of Fujian Province2016J01449
Subject Keywords:mesangial cell. Store-operated Ca2+ entry, collagen IV extracellular matrix. Smad1
Issue or Number:6
Record Number:CaltechAUTHORS:20170320-081110533
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Official Citation:Negative regulation of Smad1 pathway and collagen IV expression by store-operated Ca2+ entry in glomerular mesangial cells Peiwen Wu, Yuezhong Ren, Yuhong Ma, Yanxia Wang, Hui Jiang, Sarika Chaudhari, Mark E. Davis, Jonathan E. Zuckerman, Rong Ma American Journal of Physiology - Renal Physiology Jun 2017, 312 (6) F1090-F1100; DOI: 10.1152/ajprenal.00642.2016
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:75225
Deposited By: Ruth Sustaita
Deposited On:20 Mar 2017 15:47
Last Modified:15 Nov 2021 16:31

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