Abstract Objective To investigate the influence of silencing PAX2 gene in vivo on epithelial-mesenchymal transition (EMT) of renal tubular cells in rats with renal interstitial fibrosis. Methods A total of 64 Wistar rats were anaesthetized, and unilateral ureteral obstruction (UUO) was performed to establish a rat model of renal interstitial fibrosis. The 64 rats were randomly divided into negative control and PAX2 gene silencing groups (n = 32 each). The rats in the control group were transfected with 200 μL NC-siRNA-in vivo jetPEITM solution. Those in the PAX2 gene silencing group were transfected with 200 μL PAX2-siRNA-in vivo jetPEITM solution. Each group was further divided into 4 subgroups based on the post-transfection time (3, 5, 7 and 14 days after transfection), with 8 rats in each subgroup. Renal tissue samples were harvested in each group. Real-time PCR and Western blot were used to measure the mRNA and protein expression of PAX2 in the renal cortex, as well as the mRNA and protein expression of E-cadherin and α-SMA. Results Compared with the control group, the PAX2 gene silencing group showed significantly lower mRNA and protein expression of PAX2 (P<0.05). In the two groups, the mRNA and protein expression levels of E-cadherin were gradually reduced over the time of obstruction, while those of α-SMA gradually increased. At 14 days after transfection, the PAX2 gene silencing group had significantly higher mRNA and protein expression of E-cadherin but lower mRNA and protein expression of α-SMA compared with the control group (P<0.05). Conclusions PAX2 gene silencing can significantly inhibit the process of EMT of renal tubular cells in rats with advanced fibrosis, suggesting that PAX2 gene silencing may have a therapeutic effect on renal interstitial fibrosis.
Hewitson TD. Fibrosis in the kidney: is a problem shared a problem halved?[J]. Fibrogenesis Tissue Repair, 2012, 5(Suppl 1): S14.
[2]
Picard N, Baum O, Vogetseder A, et al. Origin of renal myofibroblasts in the model of unilateral ureter obstruction in the rat[J]. Histochem Cell Biol, 2008, 130(1): 141-155.
[3]
Liu Y. New insights into epithelial-mesenchymal transition in kidney fibrosis[J]. J Am Soc Nephrol, 2010, 21(2): 212-222.
[4]
Kim MK, Maeng YI, Sung WJ, et al. The differential expression of TGF-β1, ILK and wnt signaling inducing epithelial to mesenchymal transition in human renal fibrogenesis: an immunohistochemical study[J]. Int J Clin Exp Pathol, 2013, 6(9): 1747-1758.
[5]
Xiong H, Hong J, Du W, et al. Roles of STAT3 and ZEB1 proteins in E-cadherin down-regulation and human colorectal cancer epithelial-mesenchymal transition[J]. J Biol Chem, 2012, 287(8): 5819-5832.
[6]
Satish L, Gallo PH, Baratz ME, et al. Reversal of TGF-β1 stimulation of α-smooth muscle actin and extracellular matrix components by cyclic AMP in Dupuytren's-derived fibroblasts[J]. BMC Musculoskelet Disord, 2011, 12: 113.
[7]
Li L, Wu Y, Zhang W. PAX2 re-expression in renal tubular epithelial cells and correlation with renal interstitial fibrosis of rats with obstructive nephropathy[J]. Ren Fail, 2010, 32(5): 603-611.
[8]
André ND, Silva VA, Ariza CB, et al. In vivo knockdown of CXCR4 using jetPEI/CXCR4 shRNA nanoparticles inhibits the pulmonary metastatic potential of B16 F10 melanoma cells[J]. Mol Med Rep, 2015, 12(6): 8320-8326.
[9]
Liu H, Wang Y, Wang M, et al. Fluorinated poly(propylenimine) dendrimers as gene vectors[J]. Biomaterials, 2014, 35(20): 5407-5413.
[10]
Li L, Wu Y, Wang C, et al. Inhibition of PAX2 gene expression by siRNA (polyethylenimine) in experimental model of obstructive nephropathy[J]. Ren Fail, 2012, 34(10): 1288-1296.
[11]
Lee TY, Chang CI, Lee D, et al. RNA interference-mediated simultaneous silencing of four genes using cross-shaped RNA[J]. Mol Cells, 2013, 35(4): 320-326.
[12]
Sharma R, Sanchez-Ferras O, Bouchard M. Pax genes in renal development, disease and regeneration[J]. Semin Cell Dev Biol, 2015, 44: 97-106.
[13]
Zhou TB. Signaling pathways of PAX2 and its role in renal interstitial fibrosis and glomerulosclerosis[J]. J Recept Signal Transduct Res, 2012, 32(6): 298-303.
[14]
Li L, Wu Y, Yang Y. Paired box 2 induces epithelial-mesenchymal transition in normal renal tubular epithelial cells of rats[J]. Mol Med Rep, 2013, 7(5): 1549-1554.
[15]
Yan X, Zhang Y, Zhang H, et al. Amphiphilic polyethylenimine (PEI) as highly efficient non-viral gene carrier[J]. Org Biomol Chem, 2014, 12(12): 1975-1982.
[16]
Liang W, Gong H, Yin D, et al. High-molecular-weight polyethyleneimine conjuncted pluronic for gene transfer agents[J]. Chem Pharm Bull (Tokyo), 2011, 59(9): 1094-1101.
[17]
Liu C, Liu F, Feng L, et al. The targeted co-delivery of DNA and doxorubicin to tumor cells via multifunctional PEI-PEG based nanoparticles[J]. Biomaterials, 2013, 34(10): 2547-2564.
[18]
Huang X, Hartley AV, Yin Y, et al. AAV2 production with optimized N/P ratio and PEI-mediated transfection results in low toxicity and high titer for in vitro and in vivo applications[J]. J Virol Methods, 2013, 193(2): 270-277.
[19]
Shao W, Paul A, Abbasi S, et al. A novel polyethyleneimine-coated adeno-associated virus-like particle formulation for efficient siRNA delivery in breast cancer therapy: preparation and in vitro analysis[J]. Int J Nanomedicine, 2012, 7: 1575-1586.
[20]
Thakur S, Viswanadhapalli S, Kopp JB, et al. Activation of AMP-activated protein kinase prevents TGF-β1-induced epithelial-mesenchymal transition and myofibroblast activation [J]. Am J Pathol, 2015, 185(8): 2168-2180.
[21]
Winbanks CE, Darby IA, Kelynack KJ, et al. Explanting is an ex vivo model of renal epithelial-mesenchymal transition[J]. J Biomed Biotechnol, 2011, 2011: 212819.
