miR‑195 promotes LPS‑mediated intestinal epithelial cell apoptosis via targeting SIRT1/eIF2a

Abstract

A microarray analysis of an animal model with experimental sepsis induced by caecal ligation and puncture revealed that the level of microRNA-195 (miR-195) was upregulated. However, to the best of our knowledge, the role of miR-195 in sepsis remains unknown. The present study investigated the effect of miR-195 on apoptosis in sepsis and investigated the underlying mechanism. The level of miR-195 was measured in human intestinal epithelial cells following exposure to lipopolysaccharide (LPS). cell viability and apoptosis were detected using cell counting kit-8 and flow cytometry assays. The expression levels of apoptosis-associated proteins were determined using western blot analysis. In addition, a dual-luciferase reporter assay was employed to verify the association between miR-195 and sirtuin 1 (SIRT1). Furthermore, the SIRT1 inhibitor EX527 was applied to further confirm the regulatory network of miR-195/SIRT1 in LPS-induced apoptosis. It was demonstrated that LPS significantly inhibited cell viability and promoted cell apoptosis in NcM460 cells in a dose-dependent manner. In addition, miR-195 was significantly upregulated following LPS treatment. The present results revealed that silencing miR-195 prevented apoptosis and alleviated cell injury in LPS-induced NcM460 cells. Further investigation demonstrated that miR-195 bound directly to and negatively regulated SIRT1. Inhibition of SIRT1 reversed the protective effects of miR-195-silencing on the apoptosis and viability of NcM460 cells. Furthermore, silencing miR-195 prevented endoplasmic reticulum (ER) stress-induced apoptosis via a downregulation of SIRT1 and its downstream effectors, including activating transcription factor 4, c/EBP homologous protein, glucose-regulated protein 78 and growth arrest and dNA-damage protein 34, as well as the phosphorylation of eukaryotic translation initiation factor 2A. In conclusion, the present study revealed a novel mechanism by which miR-195 regulates SIRT1-mediated downstream effectors in ER stress-induced apoptosis in sepsis.

Effect of miR-195 inhibition on human skeletal muscle-derived stem/progenitor cells

“…The overexpression of miR-195-5p can inhibit the YAP-mediated Wnt/β-catenin signaling pathway and promote cell apoptosis, so it may be a potential therapeutic target for cancer. Zhao et al 's results [24] revealed that silencing of miR-195 prevented apoptosis and alleviated cell injury in LPS-induced NCM460 cells [25]. In our study, we observed slight (but statistically important) differences between the levels of apoptosis in the WT SkMDS/PCs population vs. SkMDS/PCs treated with miR-195 inhibitor (Figure 4), which were in favor of the "native" wild-type cells.…”

The Effects of Different Histone Deacetylase Inhibitors on Hypoxia-induced Intestinal Epithelial Barrier Dysfunction and a Preliminary Exploration of the Mechanisms

“…Inhibition of HDAC promotes acetylation at speci c lysine residues on histones and other nuclear and cytoplasmic proteins, which in turn modulates pivotal cellular functions such as chromosome remodeling, gene transcription, and cell proliferation 45,46 . It has previously been reported that HDAC inhibitors such as MS-275 25 , MC-1568 26 , Tub-A 24 , EX-527 27,28 , SAHA [29][30][31] , and VPA 23,26,32 can protect intestinal epithelial cells, improve intestinal tight junctions, support intestinal structure and function, and prevent bacterial translocation.…”

Sulforaphane protects intestinal epithelial cells against lipopolysaccharide-induced injury by activating the AMPK/SIRT1/PGC-1ɑ pathway

“…These epithelial cells are sensitive to inflammatory processes and to ROS levels that exceed the total antioxidant capacity [ 30 , 31 ]. LPS-induced injury to the intestinal tissue is also associated with increased apoptosis [ 32 ]. Consistent with our results, sulforaphane has been shown to protect the gastrointestinal mucosa from oxidative injury and inflammation induced by H. pylori and non-steroidal anti-inflammatory drugs [ 33 ].…”