Epithelial-mesenchymal Transition Of Peritoneal Mesothelial Cells Research Articles

Regulation of epithelial-mesenchymal transition via sonic hedgehog/glioma-associated oncogene homolog 1 signaling pathway in peritoneal mesothelial cells.

Sonic hedgehog (Shh) signaling regulating epithelial-mesenchymal transition (EMT) in cultured rat peritoneal mesothelial cells (PMCs) remains an under-investigated topic. The current study aimed to elucidate the role of Shh signaling in the regulation of EMT in PMCs to attenuate peritoneal injury, with the view of enhancing the efficacy of peritoneal dialysis (PD). PMCs were initially extracted from male Wistar rats using pancreatic enzyme digestion. The expression of Shh and glioma-associated oncogene homolog (Gli1) was quantitatively analyzed using the reverse-transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. Migration of PMCs was determined using Transwell assay. The expression of Shh, Gli1, and EMT markers including α-smooth muscle actin (α-SMA), fibronectin, collagen I, snail1, and E-cadherin was examined by RT-qPCR, western blot analysis, and immunofluorescence respectively. High glucose induction was identified to promote cell migration and increase the expression of Shh and Gli1 in a dose- and time-dependent manner in rat PMCs. Cyclopamine (CPN) was observed to block the Shh signaling induced by high glucose, accompanied by cell migration inhibition, decreased expression of α-SMA, fibronectin, collagen I and snail1 as well as increased expression of E-cadherin. Altogether, overexpression of Gli1 by transfected Gli1 plasmid promotes cell migration and upregulates α-SMA, fibronectin, Snail1, and collagen I expression, while downregulating E-cadherin expression. Shh/Gli1 signaling is important in mediating EMT in rat PMCs, which provides a potential novel therapeutic approach for clinical investigation on renal failure treatment.

UA promotes epithelial‑mesenchymal transition in peritoneal mesothelial cells.

Long‑term peritoneal dialysis is often limited or interrupted due to the development and progression of peritoneal fibrosis. Accumulating evidence suggests that epithelial‑mesenchymal transition (EMT) is a major component of peritoneal injury associated with peritoneal fibrosis in the end stage of renal disease; however, at present, the underlying mechanisms remain unclear. Thus, in the present study, uric acid (UA)‑induced EMT of peritoneal mesothelial cells was investigated by western‑blot and immunofluorescence staining. The results revealed that peritoneal mesothelial cells stimulated with UA underwent EMT, as demonstrated by the decreased expression of epithelial markers (E‑cadherin) and an increased expression of mesenchymal markers (α‑smooth muscle actin and vimentin). Additionally, it was reported that UA could facilitate the progression of EMT of peritoneal mesothelial cells via EMT transcription pathways, including transforming growth factor‑β1/mothers against decapentaplegic homolog 3 and P38/mitogen‑activated protein kinase by western‑blot and reverse transcription semi‑quantitative polymerase chain reaction. The results of the present study suggest that UA could promote EMT and may contribute to peritoneal chronic disease. Furthermore, the data obtained suggest that the levels of blood UA may account for the development of EMT; thus, lowering the levels of blood UA may be beneficial to inhibit the occurrence and development of peritoneal fibrosis.

Zinc supplementation inhibits the high glucose‑induced EMT of peritoneal mesothelial cells by activating the Nrf2 antioxidant pathway.

The high glucose (HG)-induced epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) serves an important role in peritoneal fibrosis (PF) during peritoneal dialysis. Our previous study reported that zinc (Zn) supplementation prevented the HG-induced EMT of rat PMCs in vitro. In the present study, the role of Zn in HG-induced EMT was investigated in vivo using a rat model of PF. Additionally, the molecular mechanisms underlying HG-induced EMT were studied in human PMCs (HPMCs). In the rat model of PF, HG treatment increased the glucose transfer capacity and decreased the ultrafiltration volume. Histopathological analysis revealed peritoneal thickening, increased expression of vimentin and decreased expression of E-cadherin. ZnSO4 significantly ameliorated the aforementioned changes, whereas Zn inhibition by clioquinol significantly aggravated the effects of HG on rats. The effects of Zn on HPMCs was assessed using western blot analysis, Transwell assays and flow cytometry. It was revealed that Zn also significantly suppressed the extent of the EMT, and reduced reactive oxygen species production and the migratory ability of HG-induced HPMCs, whereas Zn inhibition by N',N',N',N'-tetrakis (2-pyridylmethyl) ethylenediamine significantly potentiated the HG-induced EMT of HPMCs. HG-stimulated HPMCs exhibited increased expression of nuclear factor-like 2 (Nrf2) in the nucleus, and total cellular NAD(P)H quinone dehydrogenase 1 (NQO1) and heme oxygenase-1 (HO-1), the target proteins of the Nrf2 antioxidant pathway. Zn supplementation further promoted nuclear Nrf2 expression, and increased the expression of target proteins of the Nrf2 antioxidant pathway, whereas Zn depletion decreased nuclear Nrf2, NQO1 and HO-1 expression compared with the HG group. In conclusion, Zn supplementation was proposed to suppress the effects of HG on the EMT by stimulating the Nrf2 antioxidant pathway and subsequently reducing oxidative stress in PMCs.

Network-based integrated analysis of omics data reveal novel players of TGF-\u03b21-induced EMT in human peritoneal mesothelial cells

Long-term peritoneal dialysis is associated with progressive fibrosis of the peritoneum. Epithelial-mesenchymal transition (EMT) of mesothelial cells is an important mechanism involved in peritoneal fibrosis, and TGF-β1 is considered central in this process. However, targeting currently known TGF-β1-associated pathways has not proven effective to date. Therefore, there are still gaps in understanding the mechanisms underlying TGF-β1-associated EMT and peritoneal fibrosis. We conducted network-based integrated analysis of transcriptomic and proteomic data to systemically characterize the molecular signature of TGF-β1-stimulated human peritoneal mesothelial cells (HPMCs). To increase the power of the data, multiple expression datasets of TGF-β1-stimulated human cells were employed, and extended based on a human functional gene network. Dense network sub-modules enriched with differentially expressed genes by TGF-β1 stimulation were prioritized and genes of interest were selected for functional analysis in HPMCs. Through integrated analysis, ECM constituents and oxidative stress-related genes were shown to be the top-ranked genes as expected. Among top-ranked sub-modules, TNFAIP6, ZC3H12A, and NNT were validated in HPMCs to be involved in regulation of E-cadherin, ZO-1, fibronectin, and αSMA expression. The present data shows the validity of network-based integrated analysis in discovery of novel players in TGF-β1-induced EMT in peritoneal mesothelial cells, which may serve as new prognostic markers and therapeutic targets for peritoneal dialysis patients.

The Mechanism of Peritoneal Fibrosis in Peritoneal Dialysis

Peritoneal fibrosis (PF) is a common morphological change in peritoneal dialysis (PD) patients. With the progression of PF, peritoneal membrane function is impaired, which leads to ultrafiltration failure. Furthermore, PF is an essential precursor condition for the development of encapsulating peritoneal sclerosis (EPS), which is the most serious complication of PD. Epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs) plays a crucial role in PF. Transforming growth factor-?1 (TGF-?1) was thought to be the main regulator of EMT in PMCs. High glucose, hypertonicity, low pH, glucose degradation products and advanced glycation end-products in PD solution were suggested to induce TGF-?1 production. In addition, chronic inflammation mediated by infiltration of immune cells and peritoneal angiogenesis also play pivotal roles for the progression of PF.