PAX2 regulates NRP-1 expression in renal tubular epithelial cellsto promote epithelial-to-mesenchymal transition and renal fibrosis

To observe the expression of neuropilin-1 (NRP-1) induced by paired-box gene 2 (PAX2) during the process of epithelial-to-mesenchymal transition (EMT) and renal fibrosis in unilateral ureteral obstruction (UUO) model in rats, and explore the mechanism of EMT induced by PAX2. Methods: The recombinant lentivirus expression vector for PAX2 was constructed and transfected into rat normal renal tubular epithelial cell line (NRK52E). The experimental cells were divided into three groups: transfection group, empty vector group, and normal group. E-cadherin and α-SMA were detected by western blot and real-time PCR. Expression of NRP-1 was detected by western blot, real-time PCR, and immunofluorescence. Sixty male Wistar rats were randomly divided into two groups: the sham-operation group (n=30) underwent left ureteral dissection, the UUO group (n=30) underwent left ureteral ligation. Post-operation on days 3, 7, 14, 21 and 28, 6 rats from each of the groups were sacrificed and the obstructed kidneys were dissected out. The histopathological changes were observed by hematoxylin-eosin and Masson staining. E-cadherin and α-SMA were detected by western blot and immunohistochemistry. Expression of NRP-1 and PAX2 were determined by western blot, immunohistochemistry, and real-time PCR. Results: Expression of NRP-1 mRNA and protein and α-SMA protein increased (P<0.05) while E-cadherin protein expression decreased (P<0.05) in the transfection group as compared to the empty vector group in vitro. In the UUO group, fibrosis was obvious, and there was decreased expression of E-cadherin protein (P<0.05) and increased expression of α-SMA protein and NRP-1 mRNA and protein (P<0.05) in comparison to the sham group. Conclusion: NRP-1 maybe mediate PAX2-induced EMT in renal tubular epithelial cells and renal fibrosis.

Download Full-text

Metformin Improves Epithelial-to-Mesenchymal Transition Induced by TGF-β1 in Renal Tubular Epithelial NRK-52E Cells via Inhibiting Egr-1

Journal of Diabetes Research ◽

10.1155/2018/1031367 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 9

Author(s):

Meiping Guan ◽

Wenqi Li ◽

Lingling Xu ◽

Yanmei Zeng ◽

Dan Wang ◽

...

Keyword(s):

Renal Fibrosis ◽

Epithelial To Mesenchymal Transition ◽

Transforming Growth Factor ◽

Mesenchymal Transition ◽

Early Growth Response ◽

Real Time Quantitative Pcr ◽

Novel Target ◽

Renal Tubular ◽

Tgf Β1 ◽

E Cadherin

The early growth response- (Egr-) 1 has been found to play a key role in organ fibrosis. Metformin has been shown to be effective in attenuating renal tubular epithelial-to-mesenchymal transition (EMT), which is involved in renal fibrosis. However, it is unknown whether metformin improves EMT via inhibiting Egr-1. In this study, rat renal tubular epithelial (NRK-52 E) cells, treated by transforming growth factor- (TGF-)β1 of 10 ng/ml with or without metformin of 1 mmol/l, were transfected by siEgr-1 or M61-Egr-1 plasmids to knock down or overexpress Egr-1, respectively. The gene and protein expressions of E-cadherin,α-SMA, fibronectin (FN), and Egr-1 were determined by real-time quantitative PCR and Western blotting, respectively. We observed that TGF-β1 significantly reduced E-cadherin expression and upregulated the expressions of FN,α-SMA, and Egr-1, which can be reversed by metformin. M61-Egr-1 transfection could exacerbate EMT, which can be reversed by metformin. Taken together, our data show that Egr-1 plays an important role in TGF-β1-induced EMT of renal tubular epithelial cells and metformin improves EMT while inhibiting Egr-1, which provides a potential novel target to combat renal fibrosis.

Download Full-text

Inhibitory Effect of Gliquidone on Epithelial-to-Mesenchymal-Transition of Renal Tubular Epithelial Cells Under Diabetic Nephropathy Mouse Model

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2022.2870 ◽

2022 ◽

Vol 12 (1) ◽

pp. 71-80

Author(s):

Ting Liu ◽

Jie Chen ◽

Yiying Ying ◽

Ling Shi ◽

Zhengyue Chen

Keyword(s):

Diabetic Nephropathy ◽

Epithelial Cells ◽

Body Mass ◽

Renal Fibrosis ◽

Epithelial To Mesenchymal Transition ◽

Inhibitory Effect ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Mesenchymal Transition ◽

Renal Tubular

This research aimed to study the inhibitory effect of Glurenorm (gliquidone) on epithelial-to-mesenchymal-transition (EMT) of renal tubular epithelial cells based on the diabetic nephropathy (DN) model. In this study, 30 specific pathogen-free (SPF) mice were selected to construct DN model and randomly rolled into groups A, B, and C, with 10 mice in each group. Low-dose, mediumdose, and high-dose Glurenorm were administered intragastrically. The results showed that the serum urea nitrogen content (7.23±0.39 mmol/L, 6.18±0.46 mmol/L) of control and C group was considerably inferior to A group (8.01±0.48 mmol/L), and the content of C group was greatly lower than controls (P < 0.05). The creatinine clearance rate (2.97±0.44 mL/min, 4.02±0.31 mL/min) of mice in control and C group was notably superior to A group (2.18±0.38 mL/min), and that of C group was obviously higher versus controls (P < 0.05). After 5 weeks of intragastric intervention by Glurenorm, the body mass of the mice in control and C group was evidently lower relative to A group, and that of C group was obviously higher versus controls (P < 0.05). Mice in control and C group were remarkably lower in body mass at the 7th week after Glurenorm intervention versus A group, and C group was relatively lower versus controls (P < 0.05). In short, EMT played an important role in promoting the occurrence and progression of renal fibrosis. Glurenorm can reduce the progression of renal fibrosis, inhibit EMT of renal tubular epithelial cells, and effectively protect kidney function.

Download Full-text

The miR-200 family regulates TGF-β1-induced renal tubular epithelial to mesenchymal transition through Smad pathway by targeting ZEB1 and ZEB2 expression

AJP Renal Physiology ◽

10.1152/ajprenal.00268.2011 ◽

2012 ◽

Vol 302 (3) ◽

pp. F369-F379 ◽

Cited By ~ 175

Author(s):

Mingxia Xiong ◽

Lei Jiang ◽

Yang Zhou ◽

Wenjing Qiu ◽

Li Fang ◽

...

Keyword(s):

Renal Fibrosis ◽

Epithelial To Mesenchymal Transition ◽

Expression Profiles ◽

Renal Tubules ◽

Dependent Manner ◽

Mesenchymal Transition ◽

Renal Tubular ◽

Novel Therapeutic Strategies ◽

Tgf Β1

Most chronic kidney injuries inevitably progress to irreversible renal fibrosis. Tubular epithelial-to-mesenchymal transition (EMT) is recognized to play pivotal roles in the process of renal fibrosis. However, a comprehensive understanding of the pathogenesis of renal scar formation and progression remains an urgent task for renal researchers. The endogenously produced microRNAs (miRNAs), proved to play important roles in gene regulation, probably regulate most genes involved in EMT. In this study, we applied microarray analysis to investigate the expression profiles of miRNA in murine interstitial fibrotic kidneys induced by unilateral ureteral obstruction (UUO). It was found that miR-200a and miR-141, two members of the miR-200 family, were downregulated at the early phase of UUO. In TGF-β1-induced tubular EMT in vitro, it was also found that the members of the miR-200 family were downregulated in a Smad signaling-dependent manner. It was demonstrated that the miR-200 family was responsible for protecting tubular epithelial cells from mesenchymal transition by target suppression of zinc finger E-box-binding homeobox (ZEB) 1 and ZEB2, which are E-cadherin transcriptional repressors. The results suggest that downregulation of the miR-200 family initiates the dedifferentiation of renal tubules and progression of renal fibrosis, which might provide important targets for novel therapeutic strategies.

Download Full-text

Long Noncoding RNA Tug1 Promotes Angiotensin II–Induced Renal Fibrosis by Binding to Mineralocorticoid Receptor and Negatively Regulating MicroR-29b-3p

Hypertension ◽

10.1161/hypertensionaha.120.16395 ◽

2021 ◽

Author(s):

Juhong Zhang ◽

Yuqing Zhang ◽

Jing Gao ◽

Meihui Wang ◽

Xiaoting Li ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Real Time ◽

Long Noncoding Rna ◽

Mineralocorticoid Receptor ◽

Renal Fibrosis ◽

Noncoding Rna ◽

Epithelial To Mesenchymal Transition ◽

Chain Reaction ◽

Mesenchymal Transition ◽

Polymerase Chain

Inappropriately activation of renin-angiotensin-aldosterone system induced renal fibrosis is characterized by partial epithelial-to-mesenchymal transition. Previously, we have indicated that miR-29b-3p in inhibiting partial epithelial-to-mesenchymal transition by negatively regulating extracellular matrix gene expression in the kidney. Despite the critical role of miR-29b-3p in fibrosis, the molecular mechanisms by which miR-29b-3p is regulated under the condition of profibrotic stimuli are largely unknown. Our aim is to search for the long noncoding RNA that mediated sponge regulatory on miR-29b-3p, and whether the long noncoding RNA could be activated by renin-angiotensin-aldosterone system and its consequent effects on renal fibrosis. Bioinformatics analysis predicted that Tug1 might directly bound to miR-29b-3p and function as a competing endogenous RNA. Dual-luciferase reporter assay, fluorescence in situ hybridization, and real-time polymerase chain reaction were performed to indicate that Tug1 interact with miR-29b-3p in a sequence-specific manner. Decreased Tug1 led to an increase in extracellular matrix measured by Western blot, and this effect was enhanced by miR-29b-3p measured by real-time polymerase chain reaction and Western blot, suggesting cross-regulation between the RNAs. Bioinformatics analysis predicted that mineralocorticoid receptor might bind to long noncoding RNA Tug1 . Notably, mineralocorticoid receptor antagonism reduced Tug1 expression in the presence or absence of angiotensin II or aldosterone measured by real-time polymerase chain reaction, and RNA immunoprecipitation assays confirmed that the mineralocorticoid receptor directly bound to Tug1 . Finally, we confirmed that Tug1 expression was enhanced in fibrotic compared with nonfibrotic human renal biopsy samples using RNA-in situ hybridization. Our findings provide novel insights into the molecular mechanism of Ang II–induced renal fibrosis and identify the Tug1 –miR-29b-3p axis as an important target of MR activation.

Download Full-text

Runt-Related Transcription Factor 1 (RUNX1) Promotes TGF-β-Induced Renal Tubular Epithelial-to-Mesenchymal Transition (EMT) and Renal Fibrosis through the PI3K Subunit p110δ

EBioMedicine ◽

10.1016/j.ebiom.2018.04.023 ◽

2018 ◽

Vol 31 ◽

pp. 217-225 ◽

Cited By ~ 37

Author(s):

Tong Zhou ◽

Maocai Luo ◽

Wei Cai ◽

Siyuan Zhou ◽

Danying Feng ◽

...

Keyword(s):

Transcription Factor ◽

Renal Fibrosis ◽

Epithelial To Mesenchymal Transition ◽

Mesenchymal Transition ◽

Related Transcription Factor ◽

Renal Tubular ◽

Transcription Factor 1

Download Full-text

SRY-Box Transcription Factor 9 (SOX9) Affects the Proliferation, Invasion and Epithelial to Mesenchymal Transition (EMT) of Intrahepatic Cholangiocarcinoma by Regulating Transforming Growth Factor β (TGFβ)/Smad Signaling

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2772 ◽

2021 ◽

Vol 11 (10) ◽

pp. 1891-1899

Author(s):

Ling Dai ◽

Yuqing Lu ◽

Lu Jiang ◽

Liping Zhu ◽

Jing Zhang ◽

...

Keyword(s):

Cell Migration ◽

Western Blot ◽

Real Time ◽

Intrahepatic Cholangiocarcinoma ◽

Real Time Pcr ◽

Migration And Invasion ◽

Sox9 Expression ◽

Smad Signaling ◽

Tumor Tissues ◽

E Cadherin

Intrahepatic cholangiocarcinoma (ICC) develops rapidly with a high malignancy. SOX9 expression is increased in several tumors. However, its expression and role in intrahepatic cholangiocarcinoma have not yet been elucidated. Real time PCR and Western blot were done to assess SOX9 expression in tumor tissues and adjacent tissues of ICC. ICC cell line QBC939 cells were separated into control group, SOX9 overexpression group and SOX9 siRNA group followed by analysis of cell survival by MTT assay, cell migration by cell scratch assay, cell invasion by transwell chamber, E-cadherin and Vimentin level by western blot, TGFβ/Smad signaling protein level by real time PCR. SOX9 level in tumor tissues was significantly increased compared to adjacent tissues (P < 0.05) and it was associated with TNM stage, tissue type and metastasis, and survival time (P < 0.05). Transfection of pcDNA3.1-SOX9 upregulated SOX9, promoted cell proliferation, migration and invasion, downregulated E-cadherin, upregulated Vimentin, TGF-β1 and Smad4 (P < 0.05). SOX9 siRNA transfection into QBC939 cells could significantly reverse the above mentioned changes (P < 0.05). SOX9 level is increased in intrahepatic cholangiocarcinoma and targeting SOX9 can inhibit cell migration and invasion, and EMT via regulating TGFβ/Smad signaling.

Download Full-text

Pentraxin 3 Activates JNK Signaling and Regulates the Epithelial-To-Mesenchymal Transition in Renal Fibrosis

Cellular Physiology and Biochemistry ◽

10.1159/000453159 ◽

2016 ◽

Vol 40 (5) ◽

pp. 1029-1038 ◽

Cited By ~ 14

Author(s):

Tung-Wei Hung ◽

Jen-Pi Tsai ◽

Shin-Huey Lin ◽

Chien-Hsing Lee ◽

Yi-Hsien Hsieh ◽

...

Keyword(s):

Cell Migration ◽

Renal Fibrosis ◽

Epithelial To Mesenchymal Transition ◽

Pentraxin 3 ◽

Mesenchymal Transition ◽

Affect Cell Viability ◽

Stage Renal Disease ◽

End Stage ◽

E Cadherin

Background/Aims: Tubulointerstitial fibrosis can lead to end-stage renal disease. Pentraxin 3 (PTX3) is an acute phase protein produced by resident and innate immunity cells. We investigated the effect of PTX3 on cultured human proximal tubular epithelial (HK-2) cells and a rat unilateral ureteral obstruction (UUO) model of renal fibrosis. Methods: Gain-of-function experiments were used to examine the effect of recombinant human PTX3 (Rh-PTX3) on HK-2 cells. Cell proliferation (MTT assay) and in vitro cell migration were measured. The levels of PTX3, p-JNK, and EMT markers were measured using immunohistochemistry, RT-PCR, and western blotting in UUO rats and HK-2 cells. Results: HK-2 cells treated with Rh PTX3 did not affect cell viability, but significantly increased cell migration. Moreover, Rh-PTX3 increased the expression of snail, slug, N-cadherin, and vimentin, decreased the expression of E-cadherin, and increased the phosphorylation of JNK. SP600126 (a specific JNK inhibitor) enhanced the effects of Rh-PTX3. Rats with UUO exhibited time-dependent increased levels of PTX3, p-JNK, and vimentin, and decreased expression of E-cadherin. Conclusions: Our results suggest that PTX3 induces cell migration via upregulation of EMT in a JNK-dependent mechanism, and highlight the role of PTX3 in the pathogenesis renal fibrosis.

Download Full-text