Endoplasmic Reticulum Stress Mediates The Response Of Goat Endometrial Stromal Cells To Lipopolysaccharide And Zearalenone

Reproductive disorders,especially the embryonic loss in the early stage causes large economic leverage.There are several causes lead to abortion such as infection by bacteria and the mycotoxin contaminated in surrounding environment and forages.Lipopolysaccharide(LPS),as the main virulence factor of Gram-negative bacteria,also known as lipoglycans and endotoxins,are large molecules consisting of a lipid and a polysaccharide composed of O-antigen,outer core and inner core joined by a covalent bond.There has been evidence that LPS causes the abortion in domestic animals.Zearalenone(ZEA)is a potent estrogenic metabolite produced by some Fusarium and Gibberella species,which is heat-stable and is found worldwide in a number of cereal crops,such as maize,barley,oats,wheat,rice,and sorghum.Zearalenone is proved the primary toxin,causing infertility,abortion or other breeding problems.It has important significance to get a further revealing and analyzing on the mechanism of the effects of LPS and ZEA on Endometrial cells.Several factors have been reported to play critical roles during embryo implantation stage,including hormones,vasoactive genes,cytokines,growth factors,and environmental factors.However,the molecular pathways involved in the preimplantation development and embryo-uterine interaction during embryo implantation remained unclear.In this study,we focus about implantation stage and study the path-physiological change on GESCs with infection by Lipopolysaccharide(LPS)and/or Zearalenone(ZEA).LPS and/or ZEA used as a model to influence oxidative stress on cells.1.Endoplasmic Reticulum Stress is Involved in Lipopolysaccharide Induced Inflammatory Response and Apoptosis in Goat Endometrial Stromal Cells.Endoplasmic reticulum(ER)stress is involved in regulating cell metabolism,apoptosis,autophagy,and survival.However,there is not enough information about the role of ER stress on Lipopolysaccharide(LPS)induced apoptosis and inflammatory cytokines secretion in uterus.In this study,we found LPS induced apoptosis and inflammation in goat endometrial stromal cells(ESC).LPS treatment inhibited the cell viability and cell proliferation.Besides,the genes associated with proliferation,such as PCNA and MKI67,were affected by LPS treatment.Moreover,LPS increased the secretion of IL-1? and IL-8,promoted the level of MYD88,Caspase1,and TRL4.While the 4-phenyl butyric acid(4-PBA)pretreatment inhibited the expression of unfolded protein response(UPR)proteins and the secretion of inflammatory cytokines in LPS-treated cells.However,blockage of IRE1 and ATF6 did not significantly reduce apoptosis and inflammatory cytokines secretion.Collectively,ER stress involved in LPS induced apoptosis and inflammatory cytokines increase in goat ESC.This study provides new insight into the function of ER stress in the pathological process.2.Knockdown ERN1 inhidits Zearalenone-Induced Proliferation Inflammatory Response and Apoptosis In Goat Endometrial Stromal Cells.Zearalenone(ZEA)is an estrogen-like toxin produced by Fusarium that is widely found in surround environmental area.ZEA has been found to cause reproductive dysfunction in female and male animals,but the underlying mechanism remains unclear.This study examined the cell proliferation,cell apoptosis,autophagy protein expression,and some inflammatory cytokines such as IL-1? and IL-8 of goat endometrial stromal cells(ESCs)induced by different concentrations of ZEA low,middle,and high.Apoptosis rate was detected by flow cytometry,Western Blot and ELISA assay were used to identify ER stress signaling pathway and some inflammatory cytokines.The results revealed that ZEA induced cell proliferations and inhibits cell apoptosis in low and middle concentrations of ZEA.While,the high concentration of ZEA achieved cell apoptosis in ESCs.Additionally,ZEA can induces ER stress protein markers such as ATF6,IRE1?,EIF2?,and ATF4.LC3 as a marker of autophagy showed up-regulation in all of the concentrations of ZEA.IL-1? and IL-8 showed down-regulation in low concentration of ZEA but middle and high concentration showed up-regulation.Knockdown ERN1 can inhibit autophagy and the main marker of ER stress.These results suggested that IRE1 pathway can reduce apoptosis protein markers of ESCs,down activated of IRE1 can block ER stress and LC3.In addition to IL-1?and IL-8 achieved up-regulation under knock down IRE1.