scholarly journals P0721PPAR-GAMMA ACTIVATION INHIBITS TGF-BETA INDUCED RENAL COMPLEMENT AND GALECTIN-3 EXPRESSION IN VIVO AND IN VITRO

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Helga Popovics ◽  
Krisztina Mikone ◽  
Miklos Mozes ◽  
JiYeon Kwon ◽  
Georg Hansmann ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Mrna Expression ◽  
Oral Treatment ◽  
Statistical Significance ◽  
Complement C3 ◽  
Pparγ Agonist ◽  
Galectin 3 ◽  
Pioglitazone Treatment ◽  
Tgf Β1

Abstract Background and Aims Reduced peroxisome proliferator-activated receptor-γ (PPARγ) activity has been observed in chronic kidney disease and fibrosis. We have recently shown that the PPARγ agonist pioglitazone dampens TGF-β induced renal pro-fibrotic transcription factors in vivo. Several studies have shown that renal complement expression is associated with fibrosis, yet the role of galectin-3 (Lgals3) remains controversial. However, the effect of PPARγ agonists on renal complement (C3, C4b) and Lgals3 expression has not been studied. We investigated how pioglitazone treatment affects TGF-β–driven renal inflammatory molecule expression in transgenic mice and in primary tubular epithelial cell (PTEC) culture. Method Ten weeks-old male C57Bl6 control (CTL) and TGF-β transgenic mice (with elevated circulating TGF-β1 level) were divided in two sets. The first set of mice received regular chow (CTL and TGFβ, n=4/group). The second set of mice were treated orally with pioglitazone (20mg/kg/day) for 5 weeks (CTL+Pio and TGFβ+Pio, n=5/group), when the kidneys were analyzed for mRNA and protein expression. PTEC were isolated from 4-weels old CTL mouse kidneys using graded sieving and characterized by immunoblot. Isolated cells were maintained in DMEM/F12 medium supplemented with 2% FBS and insulin/transferrin/selenium, and cells of passage 4-6 were used in triplicate for the experiments. Cells were treated for 24h with 10 ng/ml TGF-β and/or 5 µM pioglitazone, then mRNA expression levels were assessed. Statistical significance was verified using Mann-Whitney test. Results TGFβ kidneys depicted 13-fold and 4-fold overexpression of C3 and C4b mRNA (p<0.05), respectively, accompanied by a 1.7-fold Lgals3 expression (p<0.05). Oral treatment with the PPARγ agonist pioglitazone inhibited the TGF-β1 induced overexpression of C3, C4b, Lgals3, as well as IL-6 and CCL2 (MCP-1) mRNA (p<0.01). This was accompanied by reduced alfa-SMA (Actca2) and fibronectin mRNA expression. Renal Lgals3 showed significant correlation with alfa-SMA expression (p<0.01, see Figure). PTEC treated with TGF-β also depicted increased C3aR, alfa-SMA and collagen-1 mRNA expression by 1.7-fold, 2-fold and 2.1-fold, respectively (p<0.01), that were reduced to control levels by pioglitazone treatment (p<0.05). Conclusion Our data indicate that PPARγ activation exert strong anti-inflammatory effects in the renal tubular epithelial cells, implicating its possible therapeutic benefit by attenuating renal EMT and fibrosis in CKD patients.

scholarly journals The c-myc proto-oncogene regulates cardiac development in transgenic mice.

1990 ◽  
Vol 10 (7) ◽  
pp. 3709-3716 ◽  
Author(s):  
T Jackson ◽  
M F Allard ◽  
C M Sreenan ◽  
L K Doss ◽  
S P Bishop ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Mrna Expression ◽  
Cardiac Myocyte ◽  
Cellular Proliferation ◽  
Cardiac Development ◽  
Constitutive Expression ◽  
Transgenic Animals ◽  
Hypertrophic Growth ◽  
Myocyte Proliferation

During the maturation of the cardiac myocyte, a transition occurs from hyperplastic to hypertrophic growth. The factors that control this transition in the developing heart are unknown. Proto-oncogenes such as c-myc have been implicated in the regulation of cellular proliferation and differentiation, and in the heart the switch from myocyte proliferation to terminal differentiation is synchronous with a decrease in c-myc mRNA abundance. To determine whether c-myc can influence myocyte proliferation or differentiation, we examined the in vivo effect of increasing c-myc expression during embryogenesis and of preventing the decrease in c-myc mRNA expression that normally occurs during cardiac development. The model system used was a strain of transgenic mice exhibiting constitutive expression of c-myc mRNA in cardiac myocytes throughout development. In these transgenic mice, increased c-myc mRNA expression was found to be associated with both atrial and ventricular enlargement. This increase in cardiac mass was secondary to myocyte hyperplasia, with the transgenic hearts containing more than twice as many myocytes as did nontransgenic hearts. The results suggest that in the transgenic animals there is additional hyperplastic growth during fetal development. However, this additional proliferative growth is not reflected in abnormal myocyte maturation, as assessed by the expression of the cardiac and skeletal isoforms of alpha-actin. The results of this study indicate that constitutive expression of c-myc mRNA in the heart during development results in enhanced hyperplastic growth and suggest a regulatory role for this proto-oncogene in cardiac myogenesis.

The c-myc proto-oncogene regulates cardiac development in transgenic mice

1990 ◽  
Vol 10 (7) ◽  
pp. 3709-3716
Author(s):  
T Jackson ◽  
M F Allard ◽  
C M Sreenan ◽  
L K Doss ◽  
S P Bishop ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Mrna Expression ◽  
Cardiac Myocyte ◽  
Cellular Proliferation ◽  
Cardiac Development ◽  
Constitutive Expression ◽  
Transgenic Animals ◽  
Hypertrophic Growth ◽  
Myocyte Proliferation

During the maturation of the cardiac myocyte, a transition occurs from hyperplastic to hypertrophic growth. The factors that control this transition in the developing heart are unknown. Proto-oncogenes such as c-myc have been implicated in the regulation of cellular proliferation and differentiation, and in the heart the switch from myocyte proliferation to terminal differentiation is synchronous with a decrease in c-myc mRNA abundance. To determine whether c-myc can influence myocyte proliferation or differentiation, we examined the in vivo effect of increasing c-myc expression during embryogenesis and of preventing the decrease in c-myc mRNA expression that normally occurs during cardiac development. The model system used was a strain of transgenic mice exhibiting constitutive expression of c-myc mRNA in cardiac myocytes throughout development. In these transgenic mice, increased c-myc mRNA expression was found to be associated with both atrial and ventricular enlargement. This increase in cardiac mass was secondary to myocyte hyperplasia, with the transgenic hearts containing more than twice as many myocytes as did nontransgenic hearts. The results suggest that in the transgenic animals there is additional hyperplastic growth during fetal development. However, this additional proliferative growth is not reflected in abnormal myocyte maturation, as assessed by the expression of the cardiac and skeletal isoforms of alpha-actin. The results of this study indicate that constitutive expression of c-myc mRNA in the heart during development results in enhanced hyperplastic growth and suggest a regulatory role for this proto-oncogene in cardiac myogenesis.

scholarly journals Pyrrole-Imidazole Polyamide Targeting Transforming Growth Factor β1 Ameliorates Encapsulating Peritoneal Sclerosis

2012 ◽  
Vol 32 (4) ◽  
pp. 462-472 ◽  
Author(s):  
Kazuo Serie ◽  
Noboru Fukuda ◽  
Shigeki Nakai ◽  
Hiroyuki Matsuda ◽  
Takashi Maruyama ◽  
...  
Keyword(s):  
Growth Factor ◽  
Mrna Expression ◽  
Messenger Rna ◽  
Transforming Growth Factor ◽  
Fluorescein Isothiocyanate ◽  
Transforming Growth Factor Β1 ◽  
Vegf Mrna ◽  
Tgf Β1

ObjectiveEncapsulating peritoneal sclerosis (EPS) is a devastating fibrotic complication in patients treated with peritoneal dialysis (PD). Transforming growth factor β1 (TGF-β1) is a pivotal factor in the induction of EPS.MethodsTo develop pyrrole-imidazole (PI) polyamide, a novel gene silencer, targeted to the TGF-β1 promoter (Polyamide) for EPS, we examined the effects of Polyamide on messenger RNA (mRNA) expression of TGF-β 1, vascular endothelial growth factor (VEGF), and extracellular matrix (ECM) in mesothelial cells in vitro, and on the thickness of injured peritoneum evaluated by histology and high- resolution regional elasticity mapping in rats in vivo.ResultsPolyamide significantly lowered mRNA expression of TGF-β 1 and ECM in vitro. Polyamide labeled with fluorescein isothiocyanate was taken up into the injured peritoneum and was strongly localized in the nuclei of most cells. Polyamide 1 mg was injected intraperitoneally 1 or 3 times in rats receiving a daily intraperitoneal injection of chlorhexidine gluconate and ethanol (CHX) for 14 days. Polyamide significantly suppressed peritoneal thickening and the abundance of TGF-β 1 and fibronectin mRNA, but did not affect expression of VEGF mRNA in the injured peritoneum. Elasticity distribution mapping showed that average elasticity was significantly lower in Polyamide-treated rats than in rats treated solely with CHX.ConclusionsPolyamide suppressed the stiffness, ECM formation, and thickening of the injured peritoneum that occurs during EPS pathogenesis. These data suggest that PI polyamide targeted to the TGF-β 1 promoter will be a specific and feasible therapeutic strategy for patients with EPS.

scholarly journals In vivo silencing of amphiregulin by a novel effective Self-Assembled-Micelle inhibitory RNA ameliorates renal fibrosis via inhibition of EGFR signals

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Seung Seob Son ◽  
Soohyun Hwang ◽  
Jun Hong Park ◽  
Youngho Ko ◽  
Sung-Il Yun ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Adhesion Molecule ◽  
Renal Fibrosis ◽  
Smooth Muscle Actin ◽  
Intercellular Adhesion Molecule ◽  
Intercellular Adhesion Molecule 1 ◽  
Kidney Fibrosis ◽  
Self Assembled ◽  
Tgf Β1

AbstractAmphiregulin (AREG) is a transmembrane glycoprotein recently implicated in kidney fibrosis. Previously, we reported that the AREG-targeting Self-Assembled-Micelle inhibitory RNA (SAMiRNA-AREG) alleviated fibrosis by stably silencing the AREG gene, and reduced the side effects of conventional siRNA treatment of pulmonary fibrosis. However, the therapeutic effect of SAMiRNA-AREG in renal fibrosis has not been studied until now. We used two animal models of renal fibrosis generated by a unilateral ureteral obstruction (UUO) and an adenine diet (AD) to investigate whether SAMiRNA-AREG inhibited renal fibrosis. To investigate the delivery of SAMiRNA-AREG to the kidney, Cy5-labeled SAMiRNA-AREG was injected into UUO- and AD-induced renal fibrosis models. In both kidney disease models, SAMiRNA-AREG was delivered primarily to the damaged kidney. We also confirmed the protective effect of SAMiRNA-AREG in renal fibrosis models. SAMiRNA-AREG markedly decreased the UUO- and AD-induced AREG mRNA expression. Furthermore, the mRNA expression of fibrosis markers, including α-smooth muscle actin, fibronectin, α1(I) collagen, and α1(III) collagen in the UUO and AD-induced kidneys, was diminished in the SAMiRNA-AREG-treated mice. The transcription of inflammatory markers (tumor necrosis factor-α and monocyte chemoattractant protein-1) and adhesion markers (vascular cell adhesion molecule 1 and intercellular adhesion molecule 1) was attenuated. The hematoxylin and eosin, Masson’s trichrome, and immunohistochemical staining results showed that SAMiRNA-AREG decreased renal fibrosis, AREG expression, and epidermal growth factor receptor (EGFR) phosphorylation in the UUO- and AD-induced models. Moreover, we studied the effects of SAMiRNA-AREG in response to TGF-β1 in mouse and human proximal tubule cells, and mouse fibroblasts. TGF-β1-induced extracellular matrix production and myofibroblast differentiation were attenuated by SAMiRNA-AREG. Finally, we confirmed that upregulated AREG in the UUO or AD models was mainly localized in the distal tubules. In conclusion, SAMiRNA-AREG represents a novel siRNA therapeutic for renal fibrosis by suppressing EGFR signals.

scholarly journals Antiproliferative Action of Calcitonin on Lactotrophs of the Rat Anterior Pituitary Gland: Evidence for the Involvement of Transforming Growth Factor β1 in Calcitonin Action

Endocrinology ◽  
2003 ◽  
Vol 144 (5) ◽  
pp. 2164-2171 ◽  
Author(s):  
Yong Qing Wang ◽  
Ren Yuan ◽  
Ya-Ping Sun ◽  
Tae-Jin Lee ◽  
Girish V. Shah
Keyword(s):  
Cell Proliferation ◽  
Mrna Expression ◽  
Adult Female ◽  
Anterior Pituitary ◽  
Cell Populations ◽  
Female Rat ◽  
Stimulatory Action ◽  
Immunopositive Cell ◽  
Tgf Β1

Calcitonin-like pituitary peptide, which is synthesized and secreted by gonadotrophs of the rat anterior pituitary (AP) gland, is a potent inhibitor of prolactin biosynthesis and lactotroph cell proliferation. Because TGF-β1 is an autocrine inhibitor of lactotroph cell proliferation, we investigated a possibility that calcitonin (CT) interacts with TGF-β1 to inhibit lactotroph cell proliferation. The actions of CT on GGH3 cell proliferation were examined in the absence or presence of anti-TGF-β1 serum. Subsequent experiments tested the effects of CT on TGF-β1 mRNA abundance as well as TGF-β1 synthesis. The studies also tested whether the stimulatory action of CT on TGF-β1 mRNA expression involves stabilization of TGF-β1 mRNA. Finally, the experiments investigated in vivo actions of CT on TGF-β1 synthesis in the AP gland. This was accomplished by studying the changes induced by iv administered CT in TGF-β1-immunopositive cell populations of adult female rat AP glands. The results have shown that the inhibitory action of CT on proliferation of GGH3 cells was attenuated by rabbit anti-TGF-β1 serum. Moreover, CT stimulated TGF-β1 mRNA expression, as well as TGF-β1 synthesis, in a dose-dependent fashion. Stimulatory action of CT on TGF-β1 expression may be posttranscriptional, because it significantly increased TGF-β1 mRNA stability. When administered in vivo, CT significantly increased TGF-β1-immunopositive cell populations of adult female rat AP gland. Colocalization studies for prolactin and TGF-β1 suggest that CT increased TGF-β1 synthesis in lactotrophs, and possibly in nonlactotroph cell populations. These results suggest that antiproliferative action of CT on lactotrophs may, at least in part, be mediated by CT-induced TGF-β1 expression.

The Enhancing Effect of -Cyclodextrin Inclusion on -Tocotrienol-dependent Negative Growth Control of Mesothelioma Cells in a Xenograft Model

2011 ◽  
Vol 1 (12) ◽  
pp. 508
Author(s):  
T. Yano ◽  
A. Sato ◽  
M. Sekine ◽  
K. Kashiwagi ◽  
M. Ota
Keyword(s):  
Tumor Growth ◽  
Anticancer Activity ◽  
Malignant Mesothelioma ◽  
Oral Treatment ◽  
Statistical Significance ◽  
Treatment Options ◽  
Growth Control ◽  
Xenograft Model ◽  
Mrna Levels

Background: Malignant mesothelioma is an aggressive cancer with no effective treatment options. Of phytochemicals, tocotrienol (T3), a member of vitamin E, is one of the most potent anti-mesothelioma agents, but the effectiveness in vivo is quite limited, due to its low bioavailability. In this study, we investigated if the oral treatment of -T3 inclusion with -cyclodextrin (CD) could improve the bioavailability and anticancer activity of the T3.Findings: Using nude mice bearing MSTO-211H cells (a human malignant mesothelioma cell line), the effect of -T3 inclusion with -CD on -T3 level in tumor tissues, tumor growth, and its related mRNA levels were examined. The difference of tumor growth between the two groups had no statistical significance, but the latter showed a lower tendency compared with the former. In linked with this observation, the level of vascular endothelial growth factor mRNA required for in vivo tumor growth in -T3 inclusion with -CD group was lower than that in -T3 group, on the contrary, the level of -T3 level showed an opposite tendency.Conclusion: Our study demonstrated that the bioavailability of -T3 was improved by an oral administration of a novel -T3 inclusion complex with CD. Furthermore, the improvement of the bioavailability contributed to the increase of anticancer activity of -T3 in vivo.Key words: Anti-cancer agent, bioavailability, cyclodextrin, mesothelioma, tocotrienol.

scholarly journals Protection of CAPE-pNO2 Against Chronic Myocardial Ischemia by the TGF-Β1/Galectin-3 Pathway in Vivo and in Vitro

2021 ◽  
Author(s):  
Qin Wan ◽  
Liwen Zhang ◽  
Yanmin Han ◽  
Zhubo Li ◽  
Boheng Li
Keyword(s):  
Myocardial Ischemia ◽  
Caffeic Acid ◽  
Transforming Growth Factor ◽  
Morphological Changes ◽  
H9c2 Cell ◽  
Galectin 3 ◽  
Tgf Β1 ◽  
Chronic Myocardial Ischemia

Abstract Background Although it is known that caffeic acid phenethyl ester (CAPE) and its derivatives could ameliorate acute myocardial injury, their effects on chronic myocardial ischemia (CMI) were not reported. This study aimed to investigate the potential effect of caffeic acid p-nitro phenethyl ester (CAPE-pNO2, a derivative of CAPE) on CMI and underlying mechanisms. Methods SD rats were subjected to high-fat-cholesterol-diet (HFCD) and vitamin D3, and the H9c2 cells were treated with LPS to establish CMI model, followed by the respective treatment with saline, CAPE or CAPE-pNO2.Results In vivo, CAPE-pNO2 could reduce serum lipid levels, and improve impaired cardiac function and morphological changes. Data of related assays indicated that CAPE-pNO2 down-regulated the expression of transforming growth factor-β1 (TGF-β1) and galectin-3 (Gal-3). Besides, CAPE-pNO2 decreased collagen deposition, the number of apoptotic cardiomyocytes and some related downstream proteins of Gal-3 in the CMI rats. Interestingly, the effects of CAPE-pNO2 on TGF-β1, Gal-3 and other proteins expression in lung were consistent with that in heart. In vitro, CAPE-pNO2 could attenuate the fibrosis, apoptosis and inflammation by activating TGF-β1/Gal-3 pathway in LPS-induced H9c2 cell. However, CAPE-pNO2-mediated cardioprotection can be eliminated when treated with modified citrus pectin (MCP, an inhibitor of Gal-3). And in comparison, CAPE-pNO2 presented stronger effects than CAPE.Conclusion This study indicates that CAPE-pNO2 may ameliorate CMI by suppressing fibrosis, inflammation and apoptosis via the TGF-β1/Gal-3 pathway in vivo and in vitro.

Abstract 372: Regulation of Ski and Scleraxis in the Infarcted Rat Heart

2015 ◽  
Vol 117 (suppl_1) ◽  
Author(s):  
Krista L Filomeno ◽  
Sunil G Rattan ◽  
Sheri Bage ◽  
Matthew Zeglinski ◽  
Michael P Czubryt ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Wound Healing ◽  
Mrna Expression ◽  
Cardiac Fibrosis ◽  
Cardiac Fibroblasts ◽  
Scar Tissue ◽  
Sprague Dawley ◽  
Scar Area ◽  
Tgf Β1

Introduction: Coronary heart disease is causal to myocardial infarction (MI) and cardiac fibrosis. Upon ischemic myocardial injury, resident cardiac fibroblasts phenoconvert to myofibroblasts and synthesize large amounts of fibrillar collagens to produce scar tissue. Although the myofibroblast numbers are reduced in the infarct scar following the completion of wound healing, a sub-population of cells persist in the wounded area, leading to maladaptive chronic remodeling of the scar area and eventually the non-infarcted myocardium. Ski has been identified as a repressor of the TGF-β1 signaling pathway, attenuating the myofibroblast phenotype and its functional properties. Scleraxis has been implicated in canonical TGF-β1 signaling to promote collagen1α2 expression. We investigated how Ski and Scleraxis contribute to physiological and pathological wound healing in vivo. Methods: The study was carried out using 64 male Sprague-Dawley rats. The left anterior descending (LAD) coronary artery was ligated to induce a myocardial infarction. Control (sham) operated animals underwent surgery without ligation of the LAD artery. Animals were sacrificed at 2, 4, and 8 weeks post-MI and tissue collected for Western blot and qPCR studies. Results: Scleraxis mRNA expression remained at baseline at 2 and 8 weeks post-MI, but was significantly increased 4 weeks post-MI. Scleraxis protein expression was down-regulated within the scar area of infarcted hearts when compared to control samples 2 and 4 weeks post-MI. Ski mRNA expression was up-regulated within the scar area of infarcted hearts 2, 4 and 8 weeks after infarction. Conclusions: Scleraxis protein is down-regulated in myofibroblasts of the infarct scar in the chronic stages of myocardial infarction, corresponding to the maturation of the scar. At these stages of wound healing, we have previously published that Ski is up-regulated in the cytosol of these same cells. We suggest reciprocal feedback in the expression of these two proteins exists in myofibroblasts in the infarct scar. We hope to learn more about the Ski/Scleraxis feedback loop in pathological wound healing to identify novel therapeutic targets.

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