Human umbilical cord mesenchymal stem cells facilitate the up‐regulation of miR‐153‐3p, whereby attenuating MGO‐induced peritoneal fibrosis in rats

Abstract

MiRNAs contribute greatly to epithelial to mesenchymal transition (EMT) of peritoneal mesothelial cells (PMCs), which is a crucial step in peritoneal fibrosis (PF). In this study, we tried to profile whether miRNA expression differences exist after human umbilical cord mesenchymal stem cells (hUCMSCs) treatment in PF rats and investigate the possible role of miR‐153‐3p involved in anti‐EMT process. We randomly assigned 34 rats into three groups: control group (Group Control), MGO‐induced PF rats (Group MGO) and hUCMSCs‐treated rats (Group MGO + hUCMSCs). MiRNA microarrays and real‐time PCR analyses were conducted in three groups. α‐SMA, Snail1 and E‐cadherin expression were detected by Western blot. Luciferase reporter assays were used to detect the effects of miR‐153‐3p overexpression on Snai1 in rat peritoneal mesothelial cells (RPMCs). We identified differentially expressed miRNAs related to EMT, in which miR‐153‐3p demonstrated the greatest increase in Group MGO + hUCMSCs. Transient cotransfection of miR‐153‐3p mimics with luciferase expression plasmids resulted in a significant repression of Snai1 3′‐untranslated region luciferase activity in RPMCs. These studies suggest that miR‐153‐3p is a critical molecule in anti‐EMT effects of hUCMSCs in MGO‐induced PF rats. MiR‐153‐3p might exert its beneficial effect through directly targeting Snai1.

SIRT1-modified human umbilical cord mesenchymal stem cells ameliorate experimental peritoneal fibrosis by inhibiting the TGF-β/Smad3 pathway

“…SIRT1 overexpression reduces TGF-β-induced extracellular matrix expression and production; in contrast, SIRT1 knockdown promotes renal function damage and enhances renal fibrosis [25]. A similar protective effect of SIRT1 was reported by another group, the authors demonstrated that SIRT1 inhibits EMT by deacetylating Smad4 and suppresses the activation of the TGF-β pathway in human endothelial cells [26]. And a combination of stem cell therapies and biotechnology is one of the present promising fields in tissue damage and wound healing.…”

“…Passage 6 hUCMSCs (1 × 10 6 ) were stained with anti-CD34, CD45, CD90, and CD105 antibodies (1:500; BD Biosciences, San Diego, USA) for 30 min at room temperature. Flow cytometry (Gilson, Middleton, WI, USA) was used to analyze antibody binding and cell surface markers' expression as described previously [26].…”

“…Male Wistar rats with 180-220 g body weight were ordered from Shanghai SLAC laboratory Animal Co., Ltd. (Shanghai, China) and used to create the peritoneal fibrosis model by intraperitoneal injection 100 mL/kg of peritoneal dialysate fluid (PDF) with 20 mmol/L MGO, following the protocol established by Li et al [26]; 8 rats were injected vehicles as the health control. After 2 weeks of PDF daily injection, the rats were divided into three groups (n = 8 each group): one group of rats were intravenously injected 1 × 10 7 hUCMSCs, another group of rats were intravenously injected 1 × 10 7 SIRT1-modified hUCMSCs, and PBS was injected into the third group as positive control of peritoneal fibrosis.…”

“…The dialysate creatinine concentration (D), plasma creatinine concentration (P), PDF glucose concentration (D0), and dialysate fluid glucose concentration (D2) were measured by using the fully automatic biochemistry analyzer (Labomed, Inc., Los Angeles, USA). The ultrafiltration volume of each rat was measured and calculated as described previously [26]. The peritoneum of each rat was collected for the subsequent analyses.…”

MicroRNA-145 promotes the epithelial-mesenchymal transition in peritoneal dialysis-associated fibrosis by suppressing fibroblast growth factor 10

“…Moreover, both miR-199a-5p and miR-214-3p were found to target claudin-2 and E-cadherin (E-cad) mRNAs to promote high glucose-induced peritoneal fibrosis in PD (12). In addition, human umbilical cord mesenchymal stem cells facilitate the up-regulation of miR-153-3p, which is a critical molecule in attenuating methylglyoxal-induced peritoneal fibrosis in rats (13). Specifically, miR-21 was shown to promote the progression of peritoneal fibrosis through activating the TGF-␤/Smad signaling pathway in in vivo and in vitro experiments (14).…”