0.05),the escape latency was longer(P

Objective To evaluate the effect of DHA on sevoflurane-induced activation of microglia.Methods N9 mouse microglia were seeded in culture plates (1 ml) or culture dishes (10 ml) at a density of 1 × 106 cells/ml and divided into 4 groups (n =18 each) using a random number table method:control group (C group),DHA group,sevoflurane group (Sevo group) and DHA plus sevoflurane group (DHA+Sevo group).Group C received no treatment.Cells were incubated in the culture medium containing 25 μmol/L DHA in DHA and DHA+Sevo groups.Cells were exposed to 2％ sevoflurane in Sevo and DHA +Sevo groups.At 24 h of culture,activated microglia were detected and counted by immunohistochemistry,the rate of CD11b positive cells was calculated,the expression of microglial biomarker CD1lb was detected by Western blot,and the concentrations of tumor necrosis factor-alpha (TNF-α) and interleukin1 beta (IL-1β) were determined by enzyme-linked immunosorbent assay.Results Compared with C and DHA groups,the rate of C D 11 b positive cells was significantly increased,the expression of CD11 b was upregulated,and the concentrations of TNF-α and IL-1β were increased in Sevo group (P＜0.05).Compared with Sevo group,the rate of CD11b positive cells was significantly decreased,the expression of CD11b was down-regulated,and the concentrations of TNF-α and IL-1β were decreased in DHA + Sevo group (P＜0.05).Conclusion DHA can decrease inflammatory responses through reducing sevoflurane-induced activation of microglia.

Objective To explore the effects of repressor element 1-silencing transcription factor (REST)/REST coinhibitory factor (CoREST) on axonal regeneration and repairmen of mouse cortical neurons after traumatic brain injury (TBI).Methods (1) The primary cortical neurons were obtained from fetal C57BL/6 mice;one,three,5,7,9,and 11 d after cultivation,miR-124 expression was detected by quantitative-(q-) PCR.(2) Neurons cultured for 5 d were divided into miR-124 mimics group,blank control group,and miR-124 inhibitor group,and miR-124 mimics,nonsense control sequences and miR-124 inhibitor were transfected,respectively;0,6,12,24,48,and 72 h after transfection,miR-124 expression was detected by q-PCR.(3) Neurons cultured for 7 d were divided into blank control group Ⅰ,oxygen glucose deprivation (OGD) model group,up-regulated miR-124+OGD model group,and down-regulated miR-124+OGD model group,and neurons in the later two groups were transfected with miR-124 mimics and miR-124 inhibitor;48 h after transfection,OGD models in the later three groups were prepared;0,6,12,24,48,and 72 h after OGD,miR-124 expression was detected by q-PCR;GAP-43,REST and CoREST expressions were detected by Western blotting 48 h after OGD;the REST and CoREST expressions were measured by immunofluorescent staining 48 h after OGD.Results (1) One,three,5,and 7 d after cultivation,miR-124 expression gradually increased,and 7,9,and 11 d after cultivation,miR-124 expression gradually decreased,with significant differences (P＜0.05).(2) Twenty-four and 48 h after transfection,miR-124 expression in the miR-124 inhibitor group was significantly lower than that in the blank control group (P＜0.05);12,24,48 and 72 h after transfection,miR-124 expression in the miR-124 mimics group was significantly higher than that in the blank control group (P＜0.05),and peak level was noted at 48 h.(3) The miR-124 expression in the OGD model group was significantly higher than that in the blank control group Ⅰ at 12,24,48 and 72 h after OGD (P＜0.05),and peak level was noted at 48 h;0,6,12,24,48,and 72 h after OGD,the miR-124 expression in the up-regulated miR-124+OGD model group was significantly higher than that in the blank control group Ⅰ (P＜0.05),and peak level was noted at 48 h;Western blotting indicated that GAP-43 and CoREST gradually increased,and REST gradually decreased in blank control group Ⅰ,OGD model group and down-regulated miR-124+OGD model group,with significant differences (P＜0.05);neurons in the up-regulated miR-124+OGD model group had significantly lower GAP-43 and CoREST expressions,and significantly higher REST expression than those in the OGD model group (P＜0.05);the results of immunofluorescence staining were consistent with those of Western blotting.Conclusion REST/CoREST,as a pair regulator,may play a key role in the repairment and regeneration of neuron axons after TBI.

Objective To investigate the protective effect of spermidine against lipopolysaccharide(LPS)-induced myocardial injury in mice and the underlying mechanism.Methods C57BL/6 mice subjected to intraperitoneal LPS injection with or without pretreatment with daily gavage of spermidine for 2 weeks were examined for myocardial pathologies using HE staining and transmission electron microscopy.In the cell experiment,cultured rat cardiomyocytes(H9c2 cells)were pretreated with 10 or 20 μmol/L spermidine before LPS exposure for 2 h,and the changes in cell viability and levels of lactate dehydrogenase(LDH)and cardiac troponin Ⅰ(cTNI)were assessed using CCK-8 kit,LDH detection kit and ELISA,respectively.Western blotting was performed to detect the changes in the expressions of Bax,Bcl-2,cleaved caspase-3,SLC7A11 and GPX4;the changes in reactive oxygen species(ROS)and Fe2+ levels were detected using fluorescent probes,and mitochondrial membrane potential of the cells was measured using JC-1 staining.Results Treatment of the mice with LPS induced obvious myocardial and mitochondrial damages,which were significantly alleviated by pretreatment with spermidine.In H9c2 cells,LPS exposure significantly lowered the cell viability,increased LDH and cTNI levels and expressions of Bax and cleaved caspase-3 levels,decreased expressions of Bcl-2,SLC7A11 and GPX4,increased ROS production and Fe2+ level(P<0.05),and lowered mitochondrial membrane potential(all P<0.05).These effects were significantly alleviated by SPD pretreatment of the cells prior to LPS exposure.Conclusion Spermidine alleviates LPS-induced myocardial injury by suppressing cell apoptosis and inhibiting cellular ROS production and ferroptosis.

Objective To investigate the protective effect of spermidine against lipopolysaccharide(LPS)-induced myocardial injury in mice and the underlying mechanism.Methods C57BL/6 mice subjected to intraperitoneal LPS injection with or without pretreatment with daily gavage of spermidine for 2 weeks were examined for myocardial pathologies using HE staining and transmission electron microscopy.In the cell experiment,cultured rat cardiomyocytes(H9c2 cells)were pretreated with 10 or 20 μmol/L spermidine before LPS exposure for 2 h,and the changes in cell viability and levels of lactate dehydrogenase(LDH)and cardiac troponin Ⅰ(cTNI)were assessed using CCK-8 kit,LDH detection kit and ELISA,respectively.Western blotting was performed to detect the changes in the expressions of Bax,Bcl-2,cleaved caspase-3,SLC7A11 and GPX4;the changes in reactive oxygen species(ROS)and Fe2+ levels were detected using fluorescent probes,and mitochondrial membrane potential of the cells was measured using JC-1 staining.Results Treatment of the mice with LPS induced obvious myocardial and mitochondrial damages,which were significantly alleviated by pretreatment with spermidine.In H9c2 cells,LPS exposure significantly lowered the cell viability,increased LDH and cTNI levels and expressions of Bax and cleaved caspase-3 levels,decreased expressions of Bcl-2,SLC7A11 and GPX4,increased ROS production and Fe2+ level(P<0.05),and lowered mitochondrial membrane potential(all P<0.05).These effects were significantly alleviated by SPD pretreatment of the cells prior to LPS exposure.Conclusion Spermidine alleviates LPS-induced myocardial injury by suppressing cell apoptosis and inhibiting cellular ROS production and ferroptosis.