Pentosan polysulfate ameliorates fibrosis and inflammation markers in SV40 MES13 cells by suppressing activation of PI3K/AKT pathway via miR-446a-3p

Abstract

Background Renal fibrosis is a common outcome of various renal damage, including diabetic nephropathy (DN), the leading cause of end-stage renal disease. Currently, there are no effective therapies for renal fibrosis. The present study aimed to determine whether pentosan polysulphate sodium (PPS), a FDA approved medication for interstitial cystitis, protects diabetic renal fibrosis. Methods Cell viability and apoptosis were evaluated in mouse mesangial cells (SV40-MES13) after incubating with the advanced glycation end products (AGEs), which play important roles in the pathogenesis of DN. Western blot and ELISA were performed to determine the expression of transforming growth factor-beta1 (TGF-β1) and fibronectin (FN), two biomarkers of renal fibrosis, as well as interleukin-6 (IL-6) and tumor necrosis factor alpha (TNFα), two biomarkers of inflammation. The miRNA-mRNA regulatory network involved in the phosphatidylinositol 3-kinase (PI3K)/Ser and Thr Kinase (AKT) signalling was investigated by miRNA deep sequencing and validated by RT-PCR and miRNA transfection. Results AGEs significantly inhibited cell proliferation and promoted cell apoptosis, which was associated with the overexpression of TGF-β1, FN, IL-6, and TNFα. PPS almost completely reversed AGEs-induced biomarkers of fibrosis and inflammation, and significantly altered the miRNA expression profile in AGEs-treated cells. Notably, the PI3K/AKT signalling was one of the most significantly enriched pathways targeted by PPS-related differentially expressed miRNAs. PPS significantly up-regulated miR-466a-3p, which was shown to target PIK3CA, and mediated the inhibitory effect of PPS on AGEs-induced activation of PI3K/AKT pathway. Conclusions The treatment of PPS protected against AGEs-induced toxicity in SV40 MES13 cells via miR-466a-3p-mediated inhibition of PI3K/AKT pathway.

Dual specificity phosphatase 22 suppresses mesangial cell hyperproliferation, fibrosis, inflammation and the MAPK signaling pathway in diabetic nephropathy

“…Cell culture. Considering that the SV40-MES13 cells were commonly used to establish the cellular DN model according to previous studies, the same cell line was chosen in the present study (14)(15)(16)(17). SV40-MES13 was obtained from National Collection of Authenticated Cell Cultures (Shanghai, China).…”

“…The SV40-MES13 cells were seeded in six-well plates (2x10 5 cells/well) and divided into HG and LG groups. In the HG group, SV40-MES13 cells were stimulated with 25 mM D-glucose (Shanghai Aladdin Biochemical Technology Co., Ltd.) for 48 h to establish cellular DN model (14)(15)(16)(17)(18), and transfected with DUSP22 overexpression plasmid (HG-oe-DUSP22) or control plasmid (HG-oe-NC) as aforementioned. In the LG group, SV40-MES13 cells were stimulated with 5.5 mM D-glucose supplemented with 19.5 mM D-mannitol (Sigma-Aldrich; Merck KGaA) for 48 h, and transfected with DUSP22 siRNA (LG-si-DUSP22) or siRNA control (LG-si-NC) as aforementioned.…”

Pentosan Polysulfate Affords Pleotropic Protection to Multiple Cells and Tissues

“…PPS stimulates proteoglycan synthesis by bovine and ovine chondrocytes cultured in the presence or absence of IL-1, and also stimulates HA production by cultured RA and OA synoviocytes. HA has important tissue-protective properties [ 45 , 62 , 63 , 64 , 65 , 66 ].…”

“…Procedures developed to reproducibly prepare PPS from beech wood hemicellulose overcome these difficulties and facilitate production of large quantities of PPS of well-defined purity. PPS is as efficient as heparin in potentiating the mitogenic activity of acidic fibroblast growth factor aFGF, acidic FGF) on human umbilical vein endothelial cells, it regulates cytokine and inflammatory mediator production [ 65 , 66 , 68 ] and it is an anti-tumor agent in a number of cancers [ 76 , 92 , 93 ]. PPS also protects against advanced glycation end-product-induced toxicity in diabetic nephropathy by inhibition of PI3K/AKT cell signaling [ 66 ].…”

Heidihuangwan alleviates renal fibrosis in rats with 5/6 nephrectomy by inhibiting autophagy

“…Fibronectin, collagen, and α-SMA increase, eventually leading to renal failure (Zhuang et al, 2019). Multiple cellular targets and molecular pathways are involved in these stages, and the cellular targets mainly include the intrinsic renal cells such as podocytes (Xuan et al, 2021), tubular epithelial cells (Hu et al, 2022a), and thylakoid cells (Xiao et al, 2022), especially tubular epithelial cells, as interstitial myofibroblasts are the main effector cells in renal fibrosis, a large proportion of which are formed through the occurrence of EMT in tubular epithelial cells in the affected kidney (Hu et al, 2021). Molecular pathways mainly include nuclear factor-kappaB (NF-κB) (Wang et al, 2018a), TGF-β1/Smad (Loboda et al, 2016), Notch, winglesstype MMTV integration site (Wnt), Hedgehog (Edeling et al, 2016), protein kinase C (PKC)/extracellular regulated protein kinases (ERK), and PI3K/Akt (Liu et al, 2017), etc.…”