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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.

SUBMITTER: Yuan T

PROVIDER: S-EPMC6984803 | biostudies-literature | 2020 Feb

REPOSITORIES: biostudies-literature

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

Yuan Ting T Zhang Li L Yao Shuo S Deng Shuang-Ya SY Liu Ji-Qiang JQ

International journal of molecular medicine 20191218 2

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). Cel ...[more]

PMID: 31894250

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Project description:Inflammatory bowel disease (IBD), which consists of Crohn's disease (CD) and ulcerative colitis (UC), is a chronic, inflammatory disorder of the gastro-intestinal tract with unknown etiology. Current evidence suggests that intestinal epithelial cells (IECs) is prominently linked to the pathogenesis of IBD. Therefore, maintaining the intact of epithelium has potential roles in improving pathophysiology and clinical outcomes of IBD. MicroRNAs (miRNAs) act as post-transcriptional gene regulators and regulate many biological processes, including embryonal development, cell differentiation, apoptosis and proliferation. In this study, we found that miR-200b decreased significantly in inflamed mucosa of IBD, especially for UC, when compared with their adjacent normal tissue. Simultaneously, we also found that the genes of E-cadherin and cyclin D1 were reduced significantly and correlated positively to the miR-200b. In addition, the upregulation of transforming growth factor-beta 1 (TGF-?1) was inversely correlated to the miR-200b in IBD. To investigate the possible roles of miR-200b in IECs maintaining, we used TGF-?1 to induce epithelial-mesenchymal transition (EMT) in IEC-6 initially. After sustained over-expressing miR-200b in IEC-6, the EMT was inhibited significantly that was characterized by downregulation of vimentin and upregulation of E-cadherin. Furthermore, we found that miR-200b enhanced E-cadherin expression through targeting of ZEB1, which encode transcriptional repressors of E-cadherin. SMAD2 was found to act as a target of miR-200b with direct evidence that miR-200b binding to the 3' UTR of SAMD2 and the ability of miR-200b to repress SMAD2 protein expression. With SMAD2 depletion, the expression of vimentin decreased correspondingly, which suggested miR-200b might reduce vimentin through regulating the SMAD2. With endogenous over-expression of miR-200b, the proliferation of IEC-6 cells increased significantly by increasing S-phase entry and promoting expression of the protein cyclin D1. Summarily, our study suggested a potential role for mir-200b in maintaining intact of intestinal epithelium through inhibiting EMT and promoting proliferation of IECs.

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MiR?195 promotes LPS?mediated intestinal epithelial cell apoptosis via targeting SIRT1/eIF2a.

Publications

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

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