Effects of CUL1 Gene on MPP+-induced SH-SY5Y Cell Survival and NLRP3 Inflammasome Pathway
Objective:To reveal the effect of Cullin1(CUL1)gene on 1-Methyl-4-phenylpyridinium ion(MPP+)-induced SH-SY5Y cell survival and NLR family,pyrin domain-containing 3(NLRP3)inflammasome pathway.Methods:(1)SH-SY5Y cells were divided into NC group,NC-sh group,CUL1-sh group,NC-OE group and CUL1-OE group.The cells were transfected with the corre-sponding lentivirus using Lipofectamine 2000 reagent.(2)SH-SY5Y cells were divided into control group,MPP+group and MPP++CUL1-OE group.The MPP+group and MPP++CUL1-OE group cells were treated with 1 mmol/L MPP+for 48 hours,while the control group cells were cultured normally.Cell proliferation was detected by MTT method.Apoptosis was detected by Annexin V-FITC/PI double staining and TUNEL staining.The level of CUL1 mRNA was detected by qRT-PCR.The protein levels of CUL1,NL-RP3,apoptosis-related spot-like protein(ASC),cleaved caspase-1,interleukin(IL)-1β and IL-18 were detected by Western blot.The lev-els of IL-1β and IL-18 in cell cuiture supernatant were detected by ELISA.Results:(1)Compared with NC group and NC-sh group,the relative expression of CUL1 mRNA and protein decreased,the relative cell viability decreased,the Annexin V-FITC/PI positive rate and the positive rate of TUNEL increased,the relative expression ofNLRP3,ASC,cleaved caspase-1,IL-1β and IL-18 protein and IL-1β and IL-18 levels in cell culture supernatant increased in CUL1-sh group(P＜0.05).Compared with NC group and NC-OE group,the relative expression of CUL1 mRNA and protein increased,the relative cell viability increased,the Annexin V-FITC/PI positive rate and the posi-tive rate of TUNEL decreased,the relative expression of NLRP3,ASC,cleaved caspase-1,IL-1β and IL-18 protein and IL-1β and IL-18 levels in cell culture supernatant decreased in CUL1-OE group(P＜0.05).(2)Compared with control group,the relative expression of CUL1 mRNA and protein decreased,the relative cell viability decreased,the Annexin V-FITC/PI positive rate and the positive rate of TUNEL increased,the relative expression of NLRP3,ASC,cleaved caspase-1,IL-1β and IL-18 protein and IL-1β and IL-18 levels in cell culture supernatant increased in MPP+group(P＜0.05).Compared with MPP+group,the relative expression of CUL1 mRNA and protein increased,the relative cell viability increased,the Annexin V-FITC/PI positive rate and the positive rate of TUNEL decreased,the relative expression ofNLRP3,ASC,cleaved caspase-1,IL-1β and IL-18 protein and IL-1β and IL-18 levels in cell culture supernatant decreased in MPP++CUL1-OE group(P＜0.05).Conclusion:CUL1 may promote the survival of SH-SY5Y cells induced by MPP+by inhibiting the activation of NLRP3 inflammasome.
万方数据
维普
