AIM:To explore the expression of cGAS/STING signaling components in Mooren’s ulcer(MU).METHODS:Samples were obtained from ten MU patients,and eight residual corneal-scleral rings of healthy donor corneas for controls...AIM:To explore the expression of cGAS/STING signaling components in Mooren’s ulcer(MU).METHODS:Samples were obtained from ten MU patients,and eight residual corneal-scleral rings of healthy donor corneas for controls.Human corneal epithelial cells(HCECs)were used to evaluate the effect of cGAS/STING signaling pathway.Immunohistochemistr y(IHC)and Western blot were used to examine the expression of cGAS,STING,and phosphorylated interferon regulatory factor 3(p-IRF3)in MU tissues.The expression of interferon-β(IFN-β)and interferon-stimulated genes(ISGs)was quantified by real-time polymerase chain reaction(PCR)and enzymelinked immunosorbent assay(ELISA).RESULTS:The protein levels of cGAS and STING in MU samples were significantly elevated when compared with the healthy controls by Western blot and IHC.After stimulation with cGAMP,real-time PCR and ELISA showed a dramatic increase of IFN-βand ISGs(containing CXCL10,IFIT1,and IL-6)in HCECs.Moreover,HCECs treated with cGAMP was characterized by increased phosphorylation and more nuclear translocation of IRF3.Meanwhile,increased p-IRF3 was observed in MU samples via IHC and Western blot.CONCLUSION:The pronounced expression of cGAS/STING signaling components in the patients with MU and probably contribute to the onset and development of MU.展开更多
BACKGROUND Hepatitis B virus(HBV)evades the innate immunity and leads to persistent chronic infection,but the molecular mechanism is still not well known.AIM To investigate whether HBV-miR-3 is involved in HBV immune ...BACKGROUND Hepatitis B virus(HBV)evades the innate immunity and leads to persistent chronic infection,but the molecular mechanism is still not well known.AIM To investigate whether HBV-miR-3 is involved in HBV immune evasion.METHODS HBV-miR-3 agomir and antagomir were employed to verify the effectiveness of HBV-miR-3 on cGAS-Sting-IFN pathway through the experiments on relative luciferase activity,cGAS protein expression,Sting phosphorylation and interferon(IFN)production.RESULTS HBV-miR-3 down-regulates cGAS protein expression post-transcriptionally by inhibition of cGAS 3’-untranslated region(3’-UTR)activity,which results in lower Sting phosphorylation and IFN production.HBV-miR-3 antagomir rescued cGAS protein expression,Sting phosphorylation and IFN-βproduction.CONCLUSION HBV-miR-3 plays an important role in HBV immunity evasion by targeting cGAS 3’-UTR and interfering with cGAS-Sting-IFN pathway.展开更多
基金Supported by National Natural Science Foundation of China(No.81900907)the Young and Middle-Aged Scientists Research Awards Fund of Shandong Province(No.ZR2017BH004)。
文摘AIM:To explore the expression of cGAS/STING signaling components in Mooren’s ulcer(MU).METHODS:Samples were obtained from ten MU patients,and eight residual corneal-scleral rings of healthy donor corneas for controls.Human corneal epithelial cells(HCECs)were used to evaluate the effect of cGAS/STING signaling pathway.Immunohistochemistr y(IHC)and Western blot were used to examine the expression of cGAS,STING,and phosphorylated interferon regulatory factor 3(p-IRF3)in MU tissues.The expression of interferon-β(IFN-β)and interferon-stimulated genes(ISGs)was quantified by real-time polymerase chain reaction(PCR)and enzymelinked immunosorbent assay(ELISA).RESULTS:The protein levels of cGAS and STING in MU samples were significantly elevated when compared with the healthy controls by Western blot and IHC.After stimulation with cGAMP,real-time PCR and ELISA showed a dramatic increase of IFN-βand ISGs(containing CXCL10,IFIT1,and IL-6)in HCECs.Moreover,HCECs treated with cGAMP was characterized by increased phosphorylation and more nuclear translocation of IRF3.Meanwhile,increased p-IRF3 was observed in MU samples via IHC and Western blot.CONCLUSION:The pronounced expression of cGAS/STING signaling components in the patients with MU and probably contribute to the onset and development of MU.
基金Supported by National Natural Science Foundation of China,Key Project,No.82430071The Scientific Research Program of FuRong Laboratory,No.2023SK2108+2 种基金Clinical Medical Research Center for Viral Hepatitis of Hunan Province,No.2023SK4009Hunan Provincial Natural Science Foundation,No.2023JJ60440Hunan Provincial Health Commission Research Program,No.C202303088786.
文摘BACKGROUND Hepatitis B virus(HBV)evades the innate immunity and leads to persistent chronic infection,but the molecular mechanism is still not well known.AIM To investigate whether HBV-miR-3 is involved in HBV immune evasion.METHODS HBV-miR-3 agomir and antagomir were employed to verify the effectiveness of HBV-miR-3 on cGAS-Sting-IFN pathway through the experiments on relative luciferase activity,cGAS protein expression,Sting phosphorylation and interferon(IFN)production.RESULTS HBV-miR-3 down-regulates cGAS protein expression post-transcriptionally by inhibition of cGAS 3’-untranslated region(3’-UTR)activity,which results in lower Sting phosphorylation and IFN production.HBV-miR-3 antagomir rescued cGAS protein expression,Sting phosphorylation and IFN-βproduction.CONCLUSION HBV-miR-3 plays an important role in HBV immunity evasion by targeting cGAS 3’-UTR and interfering with cGAS-Sting-IFN pathway.
