Nuclear Factor κB Mediates Suppression of Canonical Transient Receptor Potential 6 Expression by Reactive Oxygen Species and Protein Kinase C in Kidney Cells

Wang, Yanxia; Ding, Min; Chaudhari, Sarika; Ding, Yanfeng; Yuan, Joseph P.; Stankowska, Dorota; He, Shuning; Krishnamoorthy, Raghu R; Cunningham, J. Thomas; Ma, Rong

doi:10.1074/jbc.m112.410357

Cited by 37 publications

(30 citation statements)

References 66 publications

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“…Human MCs were purchased from Lonza (Walkersville, MD) and cultured as described in our previous studies (1,51). Cells were growth arrested with 0.5% FBS medium during treatments.…”

Section: Methodsmentioning

confidence: 99%

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Impairment of hepatic nuclear factor-4α binding to theStim1promoter contributes to high glucose-induced upregulation of STIM1 expression in glomerular mesangial cells

Wang

Chaudhari

Ren

et al. 2015

American Journal of Physiology-Renal Physiology

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Wang Y, Chaudhari S, Ren Y, Ma R. Impairment of hepatic nuclear factor 4␣ binding to the Stim1 promoter contributes to high glucoseinduced upregulation of STIM1 expression in glomerular mesangial cells. Am J Physiol Renal Physiol 308: F1135-F1145, 2015. First published March 18, 2015 doi:10.1152/ajprenal.00563.2014.-The present study was carried out to investigate if hepatic nuclear factor (HNF)4␣ 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 HNF4␣ expression in MCs. Knockdown of HNF4␣ 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 HNF4␣ reduced expressed STIM1 protein expression in human embryonic kidney-293 cells. Furthermore, high glucose treatment did not significantly change the abundance of HNF4␣ protein in MCs but significantly attenuated HNF4␣ binding activity to the Stim1 promoter. Moreover, knockdown of HNF4␣ significantly augmented store-operated Ca 2ϩ 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 HNF4␣ significantly attenuated the fibrotic response of high glucose. These results suggest that HNF4␣ negatively regulates STIM1 transcription in MCs. High glucose increases STIM1 expression levels by impairing HNF4␣ binding activity to the Stim1 promoter, which subsequently releases Stim1 transcription from HNF4␣ repression. Since the STIM1-gated storeoperated Ca 2ϩ entry pathway in MCs has an antifibrotic effect, inhibition of HNF4␣ in MCs might be a potential therapeutic option for diabetic kidney disease. hepatic nuclear factor 4␣; stromal interacting molecule 1; storeoperated Ca 2ϩ entry; mesangial cells; high glucose; diabetic nephropathy STORE-OPERATED Ca 2ϩ entry (SOCE) via store-operated Ca 2ϩ channels (SOCs) is a ubiquitous signaling mechanism in both nonexcitable and excitable cells. This Ca 2ϩ signaling regulates diverse cellular functions ranging from cell proliferation and gene expression to cell contraction and secretion (35). Although SOCE was originally described over two decades ago, its molecular mediators were unknown until recently. By high-throughput RNA inhibition screening, two protein families, stromal interacting molecule (STIM) (27, 36) and Orai (13, 48, 59), were identified as required components of SOCE. STIM1 is a single-pass transmembrane protein located primarily in the endoplasmic reticulum (ER) membrane and functions as an ER Ca 2ϩ sensor to sense the ER luminal Ca 2ϩ concentration. Orai1 is a small plasma membrane protein and is believed to be the pore-forming unit of SOCs. Upon depletion of ER Ca 2ϩ , STIM1 aggregates and translocates to ER-plasma membrane junctions, where it physically associates and subsequently activates Orai1 and causes Ca 2ϩ entry into the cytosol (...

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“…Human MCs were purchased from Lonza (Walkersville, MD) and cultured as described in our previous studies (1,51). Cells were growth arrested with 0.5% FBS medium during treatments.…”

Section: Methodsmentioning

confidence: 99%

“…Western blot analysis was performed as described in our previous publications (1,51). Briefly, whole cell lysates or nuclear extracts were fractionated by 10% SDS-PAGE, transferred to polyvinylidene difluoride membranes, and probed with primary STIM1 or actin or lamin A/C or HNF4␣ antibody.…”

Section: Methodsmentioning

confidence: 99%

Impairment of hepatic nuclear factor-4α binding to theStim1promoter contributes to high glucose-induced upregulation of STIM1 expression in glomerular mesangial cells

Wang

Chaudhari

Ren

et al. 2015

American Journal of Physiology-Renal Physiology

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“…Immunofluorescence staining was performed 2 days after transfection using the protocol described in our previous publication. 81 The cells were incubated with fibronectin primary antibody at 1:50 in PBS plus 10% donkey serum and 0.2% Triton X-100 at 4°C overnight. The secondary antibody was goat antirabbit IgG conjugated with Alexa Fluor 568 (Invitrogen) at a concentration of 1:500 for 1 hour at room temperature.…”

Section: Immunofluorescence Cytochemistrymentioning

confidence: 99%

Store–Operated Ca2+ Channels in Mesangial Cells Inhibit Matrix Protein Expression

Wang

Davis

et al. 2015

Journal of the American Society of Nephrology

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Accumulation of extracellular matrix derived from glomerular mesangial cells is an early feature of diabetic nephropathy. Ca 2+ signals mediated by store-operated Ca 2+ channels regulate protein production in a variety of cell types. The aim of this study was to determine the effect of store-operated Ca 2+ channels in mesangial cells on extracellular matrix protein expression. In cultured human mesangial cells, activation of store-operated Ca 2+ channels by thapsigargin significantly decreased fibronectin protein expression and collagen IV mRNA expression in a dose-dependent manner. Conversely, inhibition of the channels by 2-aminoethyl diphenylborinate significantly increased the expression of fibronectin and collagen IV. Similarly, overexpression of stromal interacting molecule 1 reduced, but knockdown of calcium releaseactivated calcium channel protein 1 (Orai1) increased fibronectin protein expression. Furthermore, 2-aminoethyl diphenylborinate significantly augmented angiotensin II-induced fibronectin protein expression, whereas thapsigargin abrogated high glucose-and TGF-b1-stimulated matrix protein expression. In vivo knockdown of Orai1 in mesangial cells of mice using a targeted nanoparticle siRNA delivery system resulted in increased expression of glomerular fibronectin and collagen IV, and mice showed significant mesangial expansion compared with controls. Similarly, in vivo knockdown of stromal interacting molecule 1 in mesangial cells by recombinant adeno-associated virus-encoded shRNA markedly increased collagen IV protein expression in renal cortex and caused mesangial expansion in rats. These results suggest that store-operated Ca 2+ channels in mesangial cells negatively regulate extracellular matrix protein expression in the kidney, which may serve as an endogenous renoprotective mechanism in diabetes.

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“…Although the mechanism for the celltype-specific effects of ROS is not known, one possibility is that ROS regulation of TRPC6 is cell context dependent. In MCs, the ROS/PKC or Akt/NF-jB pathway, which downregulates TRPC6 protein expression (211,193), is predominant. However, this inhibitory pathway may not be very active in podocytes, instead the Ang II signaling pathway, which upregulates TRPC6 expression (9, 169, 231, 142), may play a major role.…”

Section: Discussionmentioning

confidence: 99%

Canonical Transient Receptor Potential 6 Channel: A New Target of Reactive Oxygen Species in Renal Physiology and Pathology

Chaudhari

2016

Antioxidants & Redox Signaling

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Significance: Regulation of Ca 2+ signaling cascade by reactive oxygen species (ROS) is becoming increasingly evident and this regulation represents a key mechanism for control of many fundamental cellular functions. Canonical transient receptor potential (TRPC) 6, a member of Ca 2+ -conductive channel in the TRPC family, is widely expressed in kidney cells, including glomerular mesangial cells, podocytes, tubular epithelial cells, and vascular myocytes in renal microvasculature. Both overproduction of ROS and dysfunction of TRPC6 channel are involved in renal injury in animal models and human subjects. Although regulation of TRPC channel function by ROS has been well described in other tissues and cell types, such as vascular smooth muscle, this important cell regulatory mechanism has not been fully reviewed in kidney cells. Recent Advances: Accumulating evidence has shown that TRPC6 is a redox-sensitive channel, and modulation of TRPC6 Ca 2+ signaling by altering TRPC6 protein expression or TRPC6 channel activity in kidney cells is a downstream mechanism by which ROS induce renal damage. Critical Issues: This review highlights how recent studies analyzing function and expression of TRPC6 channels in the kidney and their response to ROS improve our mechanistic understanding of oxidative stress-related kidney diseases. Future Directions: Although it is evident that ROS regulate TRPC6-mediated Ca 2+ signaling in several types of kidney cells, further study is needed to identify the underlying molecular mechanism. We hope that the newly identified ROS/TRPC6 pathway will pave the way to new, promising therapeutic strategies to target kidney diseases such as diabetic nephropathy. Antioxid. Redox Signal. 25, 732-748.

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Nuclear Factor κB Mediates Suppression of Canonical Transient Receptor Potential 6 Expression by Reactive Oxygen Species and Protein Kinase C in Kidney Cells

Cited by 37 publications

References 66 publications

Impairment of hepatic nuclear factor-4α binding to theStim1promoter contributes to high glucose-induced upregulation of STIM1 expression in glomerular mesangial cells

Impairment of hepatic nuclear factor-4α binding to theStim1promoter contributes to high glucose-induced upregulation of STIM1 expression in glomerular mesangial cells

Store–Operated Ca2+ Channels in Mesangial Cells Inhibit Matrix Protein Expression

Canonical Transient Receptor Potential 6 Channel: A New Target of Reactive Oxygen Species in Renal Physiology and Pathology

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