It was well known that autophagy plays an important role not only in cell homeostasis, but also in innate immunity [3–7]. Invading Selinexor ic50 bacteria could be driven to the autophagosome–lysosome pathway for degradation (‘xenophagy’) which protects the host against pathogen colonization [8, 9]. It has been reported that autophagy

is necessary for cells to restrict many pathogens such as Mycobacterium tuberculosis[7, 10], Group A Streptococcus[5], Salmonella enterica[6], Francisella tularensis[1] and Rickettsia conorii[1]. Peritoneal dialysis (PD)-related peritonitis Dactolisib represents a serious complication and is the most important cause leading to the dropout in PD patients [11]. Escherichia coli (E.coli) is the most common organism caused single-germ enterobacterial peritonitis Angiogenesis inhibitor during PD [12, 13]. It was noticed in recent years that a change in the virulence of E. coli peritonitis episodes resulted in high rates of treatment failures and even mortality [12, 13]. Lipopolysaccharide (LPS) is the biologically active constituent of endotoxins derived from the cell wall of Gram-negative bacteria [10, 14], which is a potent inducer of autophagy in many cell lines, including macrophages [10], human keratinocytes [15],

and myoblasts [16]. However, the induction of autophagy by LPS in peritoneal mesothelial cells (PMCs), which provides a nonadhesive and protective layer in the abdominal cavity against the invasion of foreign

Rho particles and injury [17], and the role of autophagy in the elimination of E. coli from PMCs have not been studied yet. The objective of present study was to investigate the autophagy induced by LPS in PMCs and its role in defense against E. coli. We were specifically interested in determining whether autophagy contributes to E.coli survival or death. Methods Materials Dulbecco’s modified Eagle’s medium/F12 (DMEM/F12) and fetal bovine serum (FBS) were purchased from Gibco BRL (Grand Island, NY, USA). Ultra-pure LPS (upLPS) from Escherichia coli (O111:B4) was obtained from Invivogen (San Diego, CA, USA). Anti-LC3, anti-TLR4 and anti-Beclin-1 were from Abcam (Cambridge, UK). Vimentin was from Boster Biological Technology (Wuhan, China). Secondary antibodies were from Cell Signaling Technology (Danvers, MA, USA). Anti-cytokeratin 18 (CK-18), 3-methyladenine (3-MA), wortmannin (Wm), monodansylcadaverine (MDC), 3-[4, 5- dimethylthiazol −2 -yl]-2, 5-diphenyltetrazolium bromide (MTT), 4’,6-Diamidino-2-phenylindole dihydrochloride (DAPI), Polymyxin B (PMB) and gentamicin were from Sigma-Aldrich Co.. Fluorescent E.coli (K-12 strain) BioParticles, Lipofectamine 2000 and Annexin V-FTIC Apoptosis Detection Kit were from Invitrogen Life Technologies (Carlsbad, CA, USA).