OBJECTIVE To investigate the role of transient receptor potential melastatin 2(TRPM2),a calcium-permeable non-selective cation channel which acts as an oxidative stress sensor,in mediating the production of pro-inflam...OBJECTIVE To investigate the role of transient receptor potential melastatin 2(TRPM2),a calcium-permeable non-selective cation channel which acts as an oxidative stress sensor,in mediating the production of pro-inflammatory IL-1βin high glucose condition.METHODS Human pro-monocytic leukemia cell U937 was purchased from ATCC and cultured in RPMI 1640(Life Technologies).Prior to high glucose(HG)stimulation,U937 cells were cultured in medium with glucose 5.5mmol·L-1 for 48 h.The cells were then incubated in high glucose concentration(30mmol·L-1)or mannitol(30mmol·L-1)for 48 h.The protein expression of TRPM2 and the production of human IL-1β were evaluated by ELISA.TRPM2 inhibitors(DPQ and AMP)and TRPM2 siRNAs were employed to further investigate the role of TRPM2 in HG-induced IL-1β production.RESULTS The TRPM2 protein expression was significantly up-regulated by 2-folds in U937 cells after the treatment of high glucose(30mmol·L-1 for 48h)(P<0.01).The production of IL-1β in U937 was also significantly increased by HG treatment and was time-and dose-dependent(10,20 or 30mmol·L-1 glucose for 24,48 or 72h)(P<0.01).The HG-induced IL-1β production in U937 could be abolished by using TRPM2 inhibitors DPQ(100μmol·L-1 for 45min)and AMP(100μmol·L-1 for 45 min)as well as by the transfection of TRPM2siRNAs(60nmol·L-1).CONCLUSION High glucose condition(such as in diabetes)might mediate pro-inflammatory environments via the modulation of TRPM2 channels on immune cells.展开更多
基金The project supported by Science and Technology Development Fund of Macao SAR(118/2012/A)Research Committee of the University of Macao〔MYRG124(Y1-L3)-ICMS12-HPM〕
文摘OBJECTIVE To investigate the role of transient receptor potential melastatin 2(TRPM2),a calcium-permeable non-selective cation channel which acts as an oxidative stress sensor,in mediating the production of pro-inflammatory IL-1βin high glucose condition.METHODS Human pro-monocytic leukemia cell U937 was purchased from ATCC and cultured in RPMI 1640(Life Technologies).Prior to high glucose(HG)stimulation,U937 cells were cultured in medium with glucose 5.5mmol·L-1 for 48 h.The cells were then incubated in high glucose concentration(30mmol·L-1)or mannitol(30mmol·L-1)for 48 h.The protein expression of TRPM2 and the production of human IL-1β were evaluated by ELISA.TRPM2 inhibitors(DPQ and AMP)and TRPM2 siRNAs were employed to further investigate the role of TRPM2 in HG-induced IL-1β production.RESULTS The TRPM2 protein expression was significantly up-regulated by 2-folds in U937 cells after the treatment of high glucose(30mmol·L-1 for 48h)(P<0.01).The production of IL-1β in U937 was also significantly increased by HG treatment and was time-and dose-dependent(10,20 or 30mmol·L-1 glucose for 24,48 or 72h)(P<0.01).The HG-induced IL-1β production in U937 could be abolished by using TRPM2 inhibitors DPQ(100μmol·L-1 for 45min)and AMP(100μmol·L-1 for 45 min)as well as by the transfection of TRPM2siRNAs(60nmol·L-1).CONCLUSION High glucose condition(such as in diabetes)might mediate pro-inflammatory environments via the modulation of TRPM2 channels on immune cells.
