Abstract
Objective To investigate the effect of remimazolam(REM)on lipopolysaccharide(LPS)in-duced neurons pyroptosis by regulating the NOD-like receptor thermal protein domain associated protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(Caspase-1)/gasdermin D(GSDMD)signaling pathway.Methods Logarithmic phase mouse hippocampal neurons HT22 cells were randomly separated into a normal control group(Ctrl group),an LPS induction group(LPS group,10 mg/L LPS),a low concentration REM group(REM low group,160 μmol/L REM),a high concentration REM group(REM high group,320 μmol/L REM)and a REM high+NLRP3 activator Nigerian antibiotic group(REM high+Nigerian antibiotic group,320 μmol/L REM+10 μmol/L Nigerian antibiotic).Except for the Ctrl group,HT22 cells in all other groups were induced by LPS.After each group was treated with medication,the CCK-8 method was applied to deter-mine the absorbancy(A value)of HT22 cells.Hoechst33342/pyridine iodide(PI)staining was applied to de-tect the level of pyroptosis rate in HT22 cells.Enzyme linked immunosorbent assay(ELISA)was applied to detect the levels of interleukin(IL)-1β,IL-6 and tumor necrosis factor-α(TNF-α)in the supernatant of HT22 cells.Real-time quantitative polymerasechain reaction was applied to detect the expression levels of NLRP3 message RNA(mRNA),Caspase-1 mRNA and GSDMD mRNA in HT22 cells.Western blot was applied to detect the expression of NLRP3,Caspase-1,GSDMD and apoptosis-associated speck-like protein containing a CARD(ASC)in HT22 cells.Results The A450 values at 48 h and 72 h in HT22 cells in the LPS group were obviously lower than those in the Ctrl group(P＜0.05),while the cell apoptosis rate,IL-1β,IL-6,and TNF-αlevels in the supernatant,NLRP3,Caspase-1,GSDMD mRNA and protein expression levels,and ASC protein expression level were obviously higher than those in the Ctrl group(P＜0.05).The A450 values at 48 h and 72 h in HT22 cells in the REM low group and REM high group were obviously higher than those in the LPS group,which in the REM high group were obviously higher than those in the REM low group,and the differ-ences were statistically significant(P＜0.05).The cell apoptosis rate,IL-1β,IL-6,and TNF-α levels in the su-pernatant,NLRP3,Caspase-1,GSDMD mRNA and protein expression levels,and ASC protein expression level in the REM low group and REM high group were obviously lower than those in the LPS group,which in the REM high group were obviously lower than those in the REM low group,and the differences were statistically significant(P＜0.05).The A450 values at 48 h and 72 h in HT22 cells in the REM high+Nigerian antibiotic group were obviously lower than those in the REM high group(P＜0.05),while the cell apoptosis rate,IL-1β,IL-6,and TNF-α levels in the supernatant,NLRP3,Caspase-1,GSDMD mRNA and protein expression levels,and ASC protein expression level in the REM high+Nigerian antibiotic group were obviously higher than those in the REM high group(P＜0.05).Conclusion REM may alleviate LPS induced neuronal pyroptosis by downregulating the NLRP3/Caspase-1/GSDMD signaling pathway.关键词
瑞马唑仑/Nod样受体蛋白3/半胱氨酸天冬氨酸蛋白水解酶-1/消皮素D信号通路/脂多糖/神经元/焦亡Key words
remimazolam/NOD-like receptor thermal protein domain associated protein 3/cysteinyl as-partate specific proteinase-1/gasdermin D signaling pathway/lipopolysaccharides/neurons/pyroptosis分类
药学
