scholarly journals Inhibition of lysyl oxidase-like 2 ameliorates folic acid-induced renal tubulointerstitial fibrosis

2020 ◽  
Author(s):  
Sung-Eun Choi ◽  
Nara Jeon ◽  
Hoon Young Choi ◽  
Hyeon Joo Jeong ◽  
Beom Jin Lim
Keyword(s):  
Folic Acid ◽  
Cancer Metastasis ◽  
Transforming Growth Factor ◽  
Lysyl Oxidase ◽  
Transforming Growth Factor Β ◽  
Treated Group ◽  
Tubulointerstitial Fibrosis ◽  
Renal Tubulointerstitial Fibrosis ◽  
Mesenchymal Transformation

AbstractTubulointerstitial fibrosis is characterized by accumulation of the extracellular matrix in the interstitium. Lysyl oxidase-like 2 (LOXL2), a member of the lysyl oxidase family, is known for promoting cancer metastasis, invasion, and stromal fibrosis in various organs. Our previous study demonstrated expression of LOXL2 in kidney podocytes and tubular epithelial cells, and the association between elevated LOXL2 and tubulointerstitial fibrosis. The present study evaluated the effect of LOXL2 inhibition using an inhibitory monoclonal antibody (AB0023) on tubulointerstitial fibrosis in a folic acid-induced tubulointerstitial fibrosis mouse model. We also evaluated the association of LOXL2 with epithelial-mesenchymal transformation related molecules in vitro using HK-2 cells. Our data demonstrate that AB0023 prevented the progression of tubulointerstitial fibrosis significantly, as determined by trichrome and picro-sirius red staining, as well as the total collagen assay. The mean expression of phosphorylated Smad2 and Smad4 was lower in the AB0023-treated group although it was not statistically significant. Following transforming growth factor-β (TGF-β) challenge, LOXL2-deficient HK-2 cells exhibited significantly lower expression of the mesenchymal markers vimentin and fibronectin than control HK-2 cells. In conclusion, LOXL2 inhibition ameliorates renal fibrosis through the TGF-β/Smad signalling pathway.

scholarly journals P1196AN INIHIBITOR OF KRUPPEL-LIKE FACTOR 5 SUPPRESSES PERITONEAL FIBROSIS IN MICE

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Kumiko Muta ◽  
Yuka Nakazawa ◽  
Yoko Obata ◽  
Hiro Inoue ◽  
Kenta Torigoe ◽  
...  
Keyword(s):  
Transforming Growth Factor ◽  
Inflammatory Responses ◽  
Retinoic Acid Receptor ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β ◽  
Treated Group ◽  
Peritoneal Fibrosis ◽  
Retinoic Acid Receptor Α ◽  
Klf5 Expression

Abstract Background and Aims We presented previously that Am80, a synthetic retinoic acid receptor α specific agonist, inhibited the expression of Krüppel-like transcription factor 5 (KLF5) and reduced peritoneal fibrosis in mice. Now, we examined further detail about the mechanism to inhibit peritoneal fibrosis. Method Peritoneal fibrosis was induced by intraperitoneal injection of chlorhexidine gluconate (CG) into peritoneal cavity of ICR mice. Am80 was administered orally for every day from the start of CG injection. After 3 weeks of treatment, peritoneal tissues were examined using serial sections by immunohistochemistry to identify what kind of cells expressed KLF5. We also examined the effect of Am80 to inhibit peritoneal fibrosis in vitro. Results While KLF5 was expressed in the thickened submesothelial area of CG injected mice, Am80 treatment reduced KLF5 expression and remarkably attenuated peritoneal thickening. The numbers of transforming growth factor β positive cells, α-smooth muscle actin (αSMA) or F4/80 positive cells were significantly decreased in Am80 treated group. KLF5 was expressed in αSMA, F4/80 or CD31 positive cells. Conclusion These results indicate the KLF5 might not only associate phenotypical differentiation from fibroblasts to myofibroblasts but also regulate inflammatory responses and angiogenesis in peritoneal fibrosis model. Am80 can suppress peritoneal fibrosis through inhibiting these mechanisms. In vitro experiments are ongoing.

Antagonistic Effects of Insulin and TGF-β3 during Chondrogenic Differentiation of Human BMSCs under a Minimal Amount of Factors

2016 ◽  
Vol 201 (2) ◽  
pp. 88-96 ◽  
Author(s):  
Emilio Satoshi Hara ◽  
Mitsuaki Ono ◽  
Yuya Yoshioka ◽  
Junji Ueda ◽  
Yuri Hazehara ◽  
...  
Keyword(s):  
Chondrogenic Differentiation ◽  
Transforming Growth Factor ◽  
Direct Interaction ◽  
Transforming Growth Factor Β ◽  
Treated Group ◽  
Minimal Amount ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Antagonistic Effects

Growth factors are crucial regulators of cell differentiation towards tissue and organ development. Insulin and transforming growth factor-β (TGF-β) have been used as the major factors for chondrogenesis in vitro, by activating the AKT and Smad signaling pathways. Previous reports demonstrated that AKT and Smad3 have a direct interaction that results in the inhibition of TGF-β-mediated cellular responses. However, the result of this interaction between AKT and Smad3 during the chondrogenesis of human bone marrow-derived stem/progenitor cells (hBMSCs) is unknown. In this study, we performed functional analyses by inducing hBMSCs into chondrogenesis with insulin, TGF-β3 or in combination, and found that TGF-β3, when applied concomitantly with insulin, significantly decreases an insulin-induced increase in mRNA levels of the master regulator of chondrogenesis, SOX9, as well as the regulators of the 2 major chondrocyte markers, ACAN and COL2A1. Similarly, the insulin/TGF-β3-treated group presented a significant decrease in the deposition of cartilage matrix as detected by safranin O staining of histological sections of hBMSC micromass cultures when compared to the group stimulated with insulin alone. Intracellular analysis revealed that insulin-induced activation of AKT suppressed Smad3 activation in a dose-dependent manner. Accordingly, insulin/TGF-β3 significantly decreased the TGF-β3-induced increase in mRNA levels of the direct downstream factor of TGF-β/Smad3, CCN2/CGTF, compared to the group stimulated with TGF-β3 alone. On the other hand, insulin/TGF-β3 stimulation did not suppress insulin-induced expression of the downstream targets TSC2 and DDIT4/REDD1. In summary, insulin and TGF-β3 have antagonistic effects when applied concomitantly, with a minimal number of factors. The application of an insulin/TGF-β3 combination without further supplementation should be used with caution in the chondrogenic differentiation of hBMSCs.

Effects of folic acid supplement on uterine prostaglandin metabolism and interleukin-2 expression on day 15 of gestation in white breed and crossbred Meishan sows

2004 ◽  
Vol 84 (1) ◽  
pp. 63-72 ◽  
Author(s):  
Frédéric Guay, J. Jacques Matte ◽  
Christiane L. Girard ◽  
Marie-France Palin, Alain Giguère ◽  
Jean-Paul Laforest
Keyword(s):  
Folic Acid ◽  
Transforming Growth Factor ◽  
Interleukin 2 ◽  
Transforming Growth Factor Β ◽  
Interferon Γ ◽  
Prostaglandin E ◽  
Cyclooxygenase 1 ◽  
White Breed ◽  
Uterine Environment

The effects of a dietary folic acid (B9) supplement on the uterine environment of nulliparous Yorkshire- Landrace (YL) and multiparous Landrace (LD) and multiparous Meishan-Landrace (ML) sows were investigated. Sows were randomly assigned to two treatments: 0 mg kg-1 and 15 mg kg-1 of B9. The supplements were given daily from the estrus preceding insemination up to slaughter on Day 15 of gestation. At slaughter, one uterine horn was used to collect the “uterine flush” and conceptuses in order to determine the uterine content of prostaglandin E2(PGE2) and F2α(PGF2α), estradiol-17β (E2) and transforming growth factor-β2 (TGF-β2), and conceptus expression of cytochrome P450aromatase (P450) and interferon γ mRNA (IFNγ). The other horn was used to determine endometrial expression of interleukin-2 (IL-2), cyclooxygenase-1 (COX1) and -2 (COX2) mRNA and to evaluate endometrial in vitro secretion of PGE2. The B9 supplement tended to decrease the uterine content of PGE2 (P < 0.08), and decrease endometrial expression of COX1 mRNA (P < 0.05). The in vitro secretion of PGE2 was reduced by B9 supplement only in YL sows (P < 0.05). The type of sow did not have any effect on the uterine content of PGE2 (P > 0.10). However, endometrial expression of COX2 mRNA was lower for YL than LD sows (P < 0.05), but there was no difference between ML and LD sows. Endometrial expression of COX1 mRNA was higher for ML than LD sows (P < 0.05). The B9 supplement tended to decrease uterine content of E2 and TGF-β2 in YL and LD sows (P < 0.07), but conceptus expression of P450 mRNA increased only for YL sows (P < 0.05). The uterine content of E2 was lower for YL than LD sows (P < 0.05). The B9 supplement decreased endometrial expression of IL-2 mRNA in LD and YL sows ( P < 0.05), but increased it in ML sows (P < 0.05). Conceptus expression of IFNγ mRNA was not affected either by B9 supplementation or the type of sow. In conclusion, the effect of B9 supplementation on the porcine uterine environment at Day 15 of gestation was influenced by sow type (genotype and/or parity). Key words: Sow, gestation, folic acid, prostaglandin, interleukin-2

Leptin is a coactivator of TGF-β in unilateral ureteral obstructive kidney disease

2007 ◽  
Vol 293 (4) ◽  
pp. F1355-F1362 ◽  
Author(s):  
Philipp Kümpers ◽  
Faikah Gueler ◽  
Song Rong ◽  
Michael Mengel ◽  
Irini Tossidou ◽  
...  
Keyword(s):  
Kidney Disease ◽  
Target Genes ◽  
Leptin Receptor ◽  
Transforming Growth Factor ◽  
Rat Kidney ◽  
Peptide Hormone ◽  
Tubulointerstitial Fibrosis ◽  
Mrna Levels ◽  
Renal Tubulointerstitial Fibrosis

Progressive tubulointerstitial fibrosis is the common end point leading to end-stage renal disease in experimental and clinical settings. Since the peptide hormone leptin is involved not only in the regulation of obesity but also in the regulation of inflammation and fibrosis, we tested the hypothesis whether leptin deficiency has an impact on tubulointerstitial fibrosis in mice. Leptin-deficient ( ob/ ob) and leptin receptor-deficient mice ( db/ db) were exposed to 14 days of unilateral ureteral obstruction (UUO). The degree of fibrosis and inflammation was compared with that in sham-operated mice by performing immunohistochemistry, quantitative PCR, and Western blotting. We found that tubulointerstitial fibrosis was significantly reduced in the obstructed kidneys of ob/ ob compared with db/ db mice or control mice. Detailed analysis of infiltrating inflammatory cells by immunohistochemistry revealed a significant reduction of CD4+ cells at 14 days after UUO in both ob/ ob and db/ db mice. In contrast, we could not detect significant differences in CD8+ cells and macrophage content. Transforming growth factor (TGF)-β mRNA levels, TGF-β-induced Smad-2/3 activation, and the upregulation of downstream target genes were significantly reduced in ob/ ob mice. In addition, we demonstrated that leptin could enhance TGF-β signaling in normal rat kidney fibroblasts in vitro. We conclude that leptin can serve as a cofactor of TGF-β activation and thus plays an important role in renal tubulointerstitial fibrosis. Therefore, selective blockade of the leptin axis might provide a therapeutic possibility to prevent or delay fibrotic kidney disease.

scholarly journals circHIPK3 Exacerbates Folic Acid-Induced Renal Tubulointerstitial Fibrosis by Sponging miR-30a

2022 ◽  
Vol 12 ◽  
Author(s):  
Yan Wu ◽  
Junjun Luan ◽  
Congcong Jiao ◽  
Shiwen Zhang ◽  
Cong Ma ◽  
...  
Keyword(s):  
Folic Acid ◽  
Renal Fibrosis ◽  
Transforming Growth Factor ◽  
Periodic Acid ◽  
Expression Patterns ◽  
Tubulointerstitial Fibrosis ◽  
Immunofluorescence Staining ◽  
Pathological Feature ◽  
Renal Tubulointerstitial Fibrosis ◽  
Tgf Β1

Renal tubulointerstitial fibrosis is a common pathological feature of progressive chronic kidney disease (CKD), and current treatment has limited efficacy. The circular RNA circHIPK3 is reported to participate in the pathogenesis of various human diseases. However, the role of circHIPK3 in renal fibrosis has not been examined. In this study, we aimed to determine whether and how circHIPK3 might participate in the pathogenesis of renal fibrosis. Mice received a peritoneal injection of folic acid (250 mg/kg). Of note, 30 days later, renal fibrosis was present on periodic acid–Schiff (PAS) and Masson staining, and mRNA and protein of profibrotic genes encoding fibronectin (FN) and collagen 1 (COL1) were increased. Renal circHIPK3 was upregulated, while miR-30a was downregulated, assessed by quantitative PCR (qPCR) and fluorescence in situ hybridization (FISH). The expression of transforming growth factor beta-1 (TGF-β1) was increased by qPCR analysis, immunoblotting, and immunofluorescence. Renal circHIPK3 negatively correlated with miR-30a, and kidney miR-30a negatively correlated with TGF-β1. Target Scan and miRanda algorithms predicted three perfect binding sites between circHIPK3 and miR-30a. We found that circHIPK3, miR-30a, and TGF-β1 colocalized in the cytoplasm of human tubular epithelial cells (HK-2 cells) on FISH and immunofluorescence staining. We transfected circHIPK3 and a scrambled RNA into HK-2 cells; miR-30a was downregulated, and the profibrotic genes such as TGF-β1, FN, and COL1 were upregulated and assessed by qPCR, immunoblotting, and immunofluorescence staining. Third, the upregulation of circHIPK3, downregulation of miR-30a, and overproduction of profibrotic FN and COL1 were also observed in HK-2 cells exposed to TGF-β1. Finally, renal biopsies from patients with chronic tubulointerstitial nephritis manifested similar expression patterns of circHIPK3, miR-30a, and profibrotic proteins, such as TGF-β1, FN, and COL1 as observed in the experimental model. A feed-forward cycle was observed among circHIPK3, miR-30a, and TGF-β1. Our results suggest that circHIPK3 may contribute to progressive renal fibrosis by sponging miR-30a. circHIPK3 may be a novel therapeutic target for slowing CKD progression.

scholarly journals Inhibition of p300/CBP-Associated Factor Attenuates Renal Tubulointerstitial Fibrosis through Modulation of NF-kB and Nrf2

2019 ◽  
Vol 20 (7) ◽  
pp. 1554 ◽  
Author(s):  
Sungjin Chung ◽  
Soojeong Kim ◽  
Mina Son ◽  
Minyoung Kim ◽  
Eun Koh ◽  
...  
Keyword(s):  
Transforming Growth Factor ◽  
Cell Damage ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β ◽  
Nuclear Factor Κb ◽  
Tubulointerstitial Fibrosis ◽  
Mrna Levels ◽  
Nuclear Factor ◽  
Associated Factor ◽  
Renal Tubulointerstitial Fibrosis

p300/CBP-associated factor (PCAF), a histone acetyltransferase, is involved in many cellular processes such as differentiation, proliferation, apoptosis, and reaction to cell damage by modulating the activities of several genes and proteins through the acetylation of either the histones or transcription factors. Here, we examined a pathogenic role of PCAF and its potential as a novel therapeutic target in the progression of renal tubulointerstitial fibrosis induced by non-diabetic unilateral ureteral obstruction (UUO) in male C57BL/6 mice. Administration of garcinol, a PCAF inhibitor, reversed a UUO-induced increase in the renal expression of total PCAF and histone 3 lysine 9 acetylation and reduced positive areas of trichrome and α-smooth muscle actin and collagen content. Treatment with garcinol also decreased mRNA levels of transforming growth factor-β, matrix metalloproteinase (MMP)-2, MMP-9, and fibronectin. Furthermore, garcinol suppressed nuclear factor-κB (NF-κB) and pro-inflammatory cytokines such as tumor necrosis factor-α and IL-6, whereas it preserved the nuclear expression of nuclear factor erythroid-derived 2-like factor 2 (Nrf2) and levels of Nrf2-dependent antioxidants including heme oxygense-1, catalase, superoxide dismutase 1, and NAD(P)H:quinone oxidoreductase 1. These results suggest that the inhibition of inordinately enhanced PCAF could mitigate renal fibrosis by redressing aberrant balance between inflammatory signaling and antioxidant response through the modulation of NF-κB and Nrf2.

scholarly journals Type of MRI contrast, tissue gadolinium, and fibrosis

2014 ◽  
Vol 307 (7) ◽  
pp. F844-F855 ◽  
Author(s):  
Catherine Do ◽  
Jeffrey L. Barnes ◽  
Chunyan Tan ◽  
Brent Wagner
Keyword(s):  
Contrast Agents ◽  
Nephrogenic Systemic Fibrosis ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Transforming Growth Factor Β ◽  
Treated Group ◽  
Mri Contrast ◽  
Cultured Fibroblasts ◽  
Low K

It has been presupposed that the thermodynamic stability constant ( Ktherm) of gadolinium-based MRI chelates relate to the risk of precipitating nephrogenic systemic fibrosis. The present study compared low- Ktherm gadodiamide with high- Ktherm gadoteridol in cultured fibroblasts and rats with uninephrectomies. Gadolinium content was assessed using scanning electron microscopy equipped with energy-dispersive X-ray spectroscopy in paraffin-embedded tissues. In vitro, fibroblasts demonstrated dose-dependent fibronectin generation, transforming growth factor-β production, and expression of activated myofibroblast stress fiber protein α-smooth muscle actin. There were negligible differences with respect to toxicity or proliferation between the two contrast agents. In the rodent model, gadodiamide treatment led to greater skin fibrosis and dermal cellularity than gadoteridol. In the kidney, both contrast agents led to proximal tubule vacuolization and increased fibronectin accumulation. Despite large detectable gadolinium signals in the spleen, skin, muscle, and liver from the gadodiamide-treated group, contrast-induced fibrosis appeared to be limited to the skin and kidney. These findings support the hypothesis that low- Ktherm chelates have a greater propensity to elicit nephrogenic systemic fibrosis and demonstrate that certain tissues are resistant to these effects.

scholarly journals Interleukin-1β Promotes Epithelial-Derived Alveolar Elastogenesis via αvβ6 Integrin-Dependent TGF-β Activation

2015 ◽  
Vol 36 (6) ◽  
pp. 2198-2216 ◽  
Author(s):  
Jiarong Wang ◽  
Lei Bao ◽  
Benli Yu ◽  
Zhaoyun Liu ◽  
Wenli Han ◽  
...  
Keyword(s):  
Transforming Growth Factor ◽  
Pulmonary Inflammation ◽  
Transforming Growth Factor Β ◽  
Pathological Feature ◽  
Discernible Effect ◽  
Morphometric Analyses ◽  
Lung Epithelial ◽  
Mesenchymal Transformation ◽  
Tgf Β1

Background/Aims: IL-1β creates persistent pulmonary inflammation accompanied by elevated transforming growth factor β (TGF-β levels and is associated with abnormal elastogenesis, which is observed in bronchopulmonary dysplasia (BPD). Although progress has been made in this field, the mechanisms underlying this process remain only partially understood. Methods: We assessed aberrant elastin localization-associated signaling in mouse pups exposed to 85% O2 treated with either IL-1Ra or 1D11, using morphometric analyses, quantitative RT-PCR, immunostaining, and ELISA. We also evaluated the derivation of elastin-producing cells using dual marker tracking. The regulatory mechanisms of IL-1β were investigated in vitro in lung epithelial and mesenchymal cells. Results: Elevated levels of IL-1β, αvβ6 and TGF-β1 were each associated with aberrant elastin production in O2-exposed lungs. IL-1Ra abolished TGF-β1 activation and αvβ6 upregulation, which occurred as a result of exposure to hyperoxia, whereas 1D11 had no discernible effect on the expression of either αvβ6 or IL-1β even following O2-exposure, suggesting that IL-1β was initially induced. Additionally, double staining revealed the presence of epithelium-derived elastin-producing cells, which was confirmed via in vitro IL-1β stress-induced epithelial-mesenchymal transformation (EMT) morphological and molecular marker changes, which may explain the altered lung elastin deposition and defective septation observed in BPD. Conclusions: These data support the hypothesis that IL-1β was initially induced by hyperoxia; αvβ6 subsequently interacted with and activated TGF-β1, acting as an epithelial/mesenchymal signaling molecule that contributed to excessive alveolar elastogenesis, the primary pathological feature of BPD.

scholarly journals Pharmacological inhibition of soluble epoxide hydrolase prevents renal interstitial fibrogenesis in obstructive nephropathy

2015 ◽  
Vol 308 (2) ◽  
pp. F131-F139 ◽  
Author(s):  
Jinu Kim ◽  
Sang Pil Yoon ◽  
Myron L. Toews ◽  
John D. Imig ◽  
Sung Hee Hwang ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epoxide Hydrolase ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Soluble Epoxide Hydrolase ◽  
Tubulointerstitial Fibrosis ◽  
Tubular Injury ◽  
Pharmacological Inhibition ◽  
Renal Tubulointerstitial Fibrosis ◽  
Anti Inflammatory

Treating chronic kidney disease (CKD) has been challenging because of its pathogenic complexity. Epoxyeicosatrienoic acids (EETs) are cytochrome P-450-dependent derivatives of arachidonic acid with antihypertensive, anti-inflammatory, and profibrinolytic functions. We recently reported that genetic ablation of soluble epoxide hydrolase (sEH), an enzyme that converts EETs to less active dihydroxyeicosatrienoic acids, prevents renal tubulointerstitial fibrosis and inflammation in experimental mouse models of CKD. Here, we tested the hypothesis that pharmacological inhibition of sEH after unilateral ureteral obstruction (UUO) would attenuate tubulointerstitial fibrosis and inflammation in mouse kidneys and may provide a novel approach to manage the progression of CKD. Inhibition of sEH enhanced levels of EET regioisomers and abolished tubulointerstitial fibrosis, as demonstrated by reduced collagen deposition and myofibroblast formation after UUO. The inflammatory response was also attenuated, as demonstrated by decreased influx of neutrophils and macrophages and decreased expression of inflammatory cytokines keratinocyte chemoattractant, macrophage inflammatory protein-2, monocyte chemotactic protein-1, TNF-α, and ICAM-1 in kidneys after UUO. UUO upregulated transforming growth factor-β1/Smad3 signaling and induced NF-κB activation, oxidative stress, tubular injury, and apoptosis; in contrast, it downregulated antifibrotic factors, including peroxisome proliferator-activated receptor (PPAR) isoforms, especially PPAR-γ. sEH inhibition mitigated the aforementioned malevolent effects in UUO kidneys. These data demonstrate that pharmacological inhibition of sEH promotes anti-inflammatory and fibroprotective effects in UUO kidneys by preventing tubular injury, downregulation of NF-κB, transforming growth factor-β1/Smad3, and inflammatory signaling pathways, and activation of PPAR isoforms. Our data suggest the potential use of sEH inhibitors in treating fibrogenesis in the UUO model of CKD.

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