Endothelial expression of fractalkine (CX3CL1) is induced by Toll-like receptor 3 signaling in cultured human glomerular endothelial cells

Hirono, Koji; Imaizumi, Tadaatsu; Aizawa, Tomomi; Watanabe, Shojiro; Tsugawa, Koji; Shiratori, Toshihiro; Kawaguchi, Shogo; Seya, Kazuhiko; Matsumiya, Tomoh; Ito, Etsuro; Tanaka, Hiroshi

doi:10.1080/14397595.2019.1682768

Cited by 25 publications

(28 citation statements)

References 31 publications

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“…Experiments using si-RNA confirmed that this response was mediated by signaling via the TLR3/IFN-β axis, which is consistent with the results of our previous studies using human MCs and GECs [4][5][6][7][8][9][10][11]. Concerning type I IFNs, our previous studies using cultured human MCs and GECs demonstrated that IFN-β, but not IFN-α, synthesized de novo following TLR3 activation, is crucial in succeeding regional inflammatory cascades [3][4][5][6][7][8][9][10][11][12]. On the other hand, we found poly IC-stimulated expression of IFN-α in podocytes remained negative in our experimental setting.…”

Section: Discussionsupporting

confidence: 89%

“…ELISA kits for IFN-α and IFN-β were purchased from PBL Assay Science (Piscataway, NJ, USA). siRNA against IFN-β is previously described [7][8][9][10][11][12]. Primer sequences are listed in Table 1.…”

Section: Reagentsmentioning

confidence: 99%

“…Since the expression of TLR3 in resident glomerular cells is confirmed in biopsy specimens from patients with CKD [4,5], regional antiviral host defenses via the activation of TLR3 signaling in the intrinsic glomerular cells have been postulated to be involved in the pathogenesis of some CKD types [4][5][6]. To date, considering the implications of TLR3 signaling in the pathogenesis of CKD, we treated cultured human mesangial cells (MCs) and glomerular endothelial cells (GECs) with polyinosinic-polycytidylic acid (poly IC), an authentic viral doublestranded RNA, and thereafter examined the expressions of representative proinflammatory chemokines and cytokines [6][7][8][9][10][11][12]. Notably, podocytes themselves also reportedly express TLR3 and its downstream signaling pathways, and these axes are presumed to be involved in the pathogenesis of certain CKD forms [13,14].…”

Section: Introductionmentioning

confidence: 99%

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Interleukin-6 via Toll-Like Receptor 3 Signaling Attenuates the Expression of Proinflammatory Chemokines in Human Podocytes

Umetsu

Watanabe

Imaizumi

et al. 2021

Kidney Blood Press Res

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Background: Although toll-like receptor 3 (TLR3) signaling is involved in the development of certain chronic kidney diseases, the specific molecular mechanisms underlying inflammatory reactions via activation of TLR3 signaling in human podocytes remain unclear. Interleukin (IL)-6 is a pleiotropic cytokine associated with innate and adaptive immune responses; however, little is known about the implication of IL-6 via the activation of regional TLR3 signaling in the inflammatory reactions in human podocytes. Methods: We treated immortalized human podocytes with polyinosinic-polycytidylic acid (poly IC), an authentic viral double-stranded RNA, and assessed the expression of IL-6, monocyte chemoattractant protein-1 (MCP-1), and C-C motif chemokine ligand 5 (CCL5) using quantitative real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. To further elucidate the poly IC-induced signaling pathway, we subjected the cells to RNA interference against IFN-β and IL-6. Results: We found that the activation of TLR3 induced expression of IL-6, MCP-1, CCL5, and IFN-β in human podocytes. RNA interference experiments revealed that IFN-β was involved in the poly IC-induced expression of IL-6, MCP-1, and CCL5. Interestingly, IL-6 knockdown markedly increased the poly IC-induced expression of MCP-1 and CCL5. Further, treatment of cells with IL-6 attenuated the expression of CCL5 and MCP-1 mRNA and proteins. Conclusion: IL-6 induced by TLR3 signaling negatively regulates the expression of representative TLR3 signaling-dependent proinflammatory chemokines in human podocytes.

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Section: Discussionsupporting

confidence: 89%

Section: Reagentsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Interleukin-6 via Toll-Like Receptor 3 Signaling Attenuates the Expression of Proinflammatory Chemokines in Human Podocytes

Umetsu

Watanabe

Imaizumi

et al. 2021

Kidney Blood Press Res

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“…Poly (I:C)/TLR3 triggers NF-κB activation and interleukin-6 (IL-6) production in astrocytes [ 16 ] and ROS-mediated NF-κB activation in macrophages [ 17 ]. Poly (I:C) induces CX3CL1 expression via activation of TLR3/NF-κB in glomerular endothelial cells [ 18 ]. These studies show the relationship between ROS and NF-κB activation or cytokine expression in cells exposed to TNF-α or poly (I:C).…”

Section: Introductionmentioning

confidence: 99%

Docosahexaenoic Acid Inhibits Cytokine Expression by Reducing Reactive Oxygen Species in Pancreatic Stellate Cells

2021

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Pancreatic stellate cells (PSCs) are activated by inflammatory stimuli, such as TNF-α or viral infection. Activated PSCs play a crucial role in the development of chronic pancreatitis. Polyinosinic-polycytidylic acid (poly (I:C)) is structurally similar to double-stranded RNA and mimics viral infection. Docosahexaenoic acid (DHA) exhibits anti-inflammatory activity. It inhibited fibrotic mediators and reduced NF-κB activity in the pancreas of mice with chronic pancreatitis. The present study aimed to investigate whether DHA could suppress cytokine expression in PSCs isolated from rats. Cells were pre-treated with DHA or the antioxidant N-acetylcysteine (NAC) and stimulated with TNF-α or poly (I:C). Treatment with TNF-α or poly (I:C) increased the expression of monocyte chemoattractant protein 1 (MCP-1) and chemokine C-X3-C motif ligand 1 (CX3CL1), which are known chemoattractants, and enhanced intracellular and mitochondrial reactive oxygen species (ROS) production and NF-κB activity, but reduced mitochondrial membrane potential (MMP). Increased intracellular and mitochondrial ROS accumulation, cytokine expression, MMP disruption, and NF-κB activation were all prevented by DHA in TNF-α-or poly (I:C)-treated PSCs. NAC suppressed TNF-α-or poly (I:C)-induced expression of MCP-1 and CX3CL1. In conclusion, DHA inhibits poly (I:C)-or TNF-α-induced cytokine expression and NF-κB activation by reducing intracellular and mitochondrial ROS in PSCs. Consumption of DHA-rich foods may be beneficial in preventing chronic pancreatitis by inhibiting cytokine expression in PSCs.

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“…More studies are therefore required to fully characterize the influence of laminar flow on GEnC, for example by studying the effect of a disturbed glomerular blood flow on the inflammatory phenotype of GEnC. GEnCs cultured under static conditions are often used as an experimental tool in research on the glomerular filtration barrier [34][35][36]. Based on the findings in the current study, culturing GEnC under laminar flow for 7 days might improve the potential of GEnC to mimic glomerular physiology in vitro and eventually a better understanding of the glomerular filtration barrier.…”

Section: Plos Onementioning

confidence: 99%

Laminar flow substantially affects the morphology and functional phenotype of glomerular endothelial cells

2021

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Shear stress induced by laminar blood flow has a profound effect on the morphology and functional phenotype of macrovascular endothelial cells. The influence of laminar flow on the glomerular microvascular endothelium, however, remains largely elusive. The glomerular endothelium, including its glycocalyx, is a crucial part of the glomerular filtration barrier, which is involved in blood filtration. We therefore investigated the influence of laminar flow-induced shear stress on the glomerular endothelium. Conditionally immortalized mouse glomerular endothelial cells were cultured for 7 days under a laminar flow of 5 dyn/cm2 to mimic the glomerular blood flow. The cells were subsequently analysed for changes in morphology, expression of shear stress-responsive genes, nitric oxide production, glycocalyx composition, expression of anti-oxidant genes and the inflammatory response. Culture under laminar flow resulted in cytoskeletal rearrangement and cell alignment compared to static conditions. Moreover, production of nitric oxide was increased and the expression of the main functional component of the glycocalyx, Heparan Sulfate, was enhanced in response to shear stress. Furthermore, glomerular endothelial cells demonstrated a quiescent phenotype under flow, characterized by a decreased expression of the pro-inflammatory gene ICAM-1 and increased expression of the anti-oxidant enzymes HO-1 and NQO1. Upon exposure to the inflammatory stimulus TNFα, however, glomerular endothelial cells cultured under laminar flow showed an enhanced inflammatory response. In conclusion, laminar flow extensively affects the morphology and functional phenotype of glomerular endothelial cells in culture. Furthermore, glomerular endothelial cells respond differently to shear stress compared to macrovascular endothelium. To improve the translation of future in vitro studies with glomerular endothelial cells to the in vivo situation, it appears therefore crucial to culture glomerular endothelial cells under physiological flow conditions.

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Endothelial expression of fractalkine (CX3CL1) is induced by Toll-like receptor 3 signaling in cultured human glomerular endothelial cells

Cited by 25 publications

References 31 publications

Interleukin-6 via Toll-Like Receptor 3 Signaling Attenuates the Expression of Proinflammatory Chemokines in Human Podocytes

Interleukin-6 via Toll-Like Receptor 3 Signaling Attenuates the Expression of Proinflammatory Chemokines in Human Podocytes

Docosahexaenoic Acid Inhibits Cytokine Expression by Reducing Reactive Oxygen Species in Pancreatic Stellate Cells

Laminar flow substantially affects the morphology and functional phenotype of glomerular endothelial cells

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