Objective:Inflammation in the central nervous system plays a crucial role in the occurrence and development of sepsis-associated encephalopathy.This study aims to explore the effects of maresin 1(MaR1),an anti-inflamm...Objective:Inflammation in the central nervous system plays a crucial role in the occurrence and development of sepsis-associated encephalopathy.This study aims to explore the effects of maresin 1(MaR1),an anti-inflammatory and pro-resolving lipid mediator,on sepsis-induced neuroinflammation and cognitive impairment.Methods:Mice were randomly assigned to 4 groups:A sham group(sham operation+vehicle),a cecal ligation and puncture(CLP)group(CLP operation+vehicle),a MaR1-LD group(CLP operation+1 ng MaR1),and a MaR1-HD group(CLP operation+10 ng MaR1).MaR1 or vehicle was intraperitoneally administered starting 1 h before CLP operation,then every other day for 7 days.Survival rates were monitored,and serum inflammatory cytokines[tumor necrosis factor alpha(TNF-α),interleukin(IL)-1β,and IL-6]were measured 24 h after operation using enzyme-linked immunosorbent assay(ELISA).Cognitive function was assessed 7 days after operation using the Morris water maze(MWM)test and novel object recognition(NOR)task.The mRNA expression of TNF-α,IL-1β,IL-6,inducible nitric oxide synthase(iNOS),IL-4,IL-10,and arginase 1(Arg1)in cortical and hippocampal tissues was determined by real-time reverse transcription PCR(RT-PCR).Western blotting was used to determine the protein expression of iNOS,Arg1,signal transducer and activator of transcription 6(STAT6),peroxisome proliferator-activated receptor gamma(PPARγ),and phosphorylated STAT6(p-STAT6)in hippocampal tissue.Microglia activation was visualized via immunofluorescence.Mice were also treated with the PPARγantagonist GW9662 to confirm the involvement of this pathway in MaR1’s effects.Results:CLP increased serum levels of TNF-α,IL-1β,and IL-6,and reduced body weight and survival rates(all P<0.05).Both 1 ng and 10 ng doses of MaR1 significantly reduced serum TNF-α,IL-1β,and IL-6 levels,improved body weight,and increased survival rates(all P<0.05).No significant difference in efficacy was observed between the 2 doses(all P>0.05).MWM test and NOR task indicated that CLP impaired spatial learning,which MaR1 mitigated.However,GW9662 partially reversed MaR1’s protective effects.Real-time RTPCR results demonstrated that,compared to the sham group,mRNA expression of TNF-α,IL-1β,and iNOS significantly increased in hippocampal tissues following CLP(all P<0.05),while IL-4,IL-10,and Arg1 showed a slight decrease,though the differences were not statistically significant(all P>0.05).Compared to the CLP group,both 1 ng and 10 ng MaR1 decreased TNF-α,IL-1β,and iNOS mRNA expression in hippocampal tissues and increased IL-4,IL-10,and Arg1 mRNA expression(all P<0.05).Immunofluorescence results indicated a significant increase in Iba1-positive microglia in the hippocampus after CLP compared to the sham group(P<0.05).Administration of 1 ng and 10 ng MaR1 reduced the percentage area of Iba1-positive cells in the hippocampus compared to the CLP group(both P<0.05).Western blotting results showed that,compared to the CLP group,both 1 ng and 10 ng MaR1 down-regulated the iNOS expression,while up-regulated the expression of Arg1,PPARγ,and p-STAT6(all P<0.05).However,the inclusion of GW9662 counteracted the MaR1-induced upregulation of Arg1 and PPARγcompared to the MaR1-LD group(all P<0.05).Conclusion:MaR1 inhibits the classical activation of hippocampal microglia,promotes alternative activation,reduces sepsis-induced neuroinflammation,and improves cognitive decline.展开更多
目的采用Meta分析的方法评价右美托咪定联合局麻药腹腔滴注用于腹腔镜手术术后镇痛的有效性和安全性。方法检索Cochrane、Embase、Web of Science、Pubmed、CBM、知网、万方和维普等数据库,纳入比较右美托咪定联合局麻药(研究组)与单独...目的采用Meta分析的方法评价右美托咪定联合局麻药腹腔滴注用于腹腔镜手术术后镇痛的有效性和安全性。方法检索Cochrane、Embase、Web of Science、Pubmed、CBM、知网、万方和维普等数据库,纳入比较右美托咪定联合局麻药(研究组)与单独局麻药(对照组)腹腔滴注的随机对照试验(RCT)。两名研究者独立进行文献筛选、质量评价、数据提取后,分别用RevMan 5.3和TSAv0.9Beta进行Meta分析和试验序贯分析(TSA)。结果共纳入5项RCT研究,共320例患者。与对照组比较,研究组术后24 h内各时点VAS疼痛评分明显降低(P<0.05),术后镇痛时间明显延长(P=0.002),镇痛药使用量明显减少(P<0.001),术后肩痛发生率明显降低(P<0.001),术后呕吐的发生率明显降低(P<0.05)。两组术后恶心、低血压和心动过缓的发生率差异无统计学意义。结论在腹腔镜手术中右美托咪定作为局麻药的佐剂腹腔滴注增强局麻药的作用效果,但鉴于该给药途径超说明书使用,腹腔内使用时应谨慎。展开更多
目的:建立吗啡耐受大鼠腰段脊髓组织蛋白质组双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)图谱,比较分析吗啡耐受大鼠腰段脊髓组织中蛋白质组的变化和差异表达,鉴定出差异表达的蛋白质点并进行验证。方法:将16只鞘内置管雄...目的:建立吗啡耐受大鼠腰段脊髓组织蛋白质组双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)图谱,比较分析吗啡耐受大鼠腰段脊髓组织中蛋白质组的变化和差异表达,鉴定出差异表达的蛋白质点并进行验证。方法:将16只鞘内置管雄性SD大鼠随机分为吗啡耐受组(MT组,n=8)和生理盐水组(NS组,n=8),取其腰段脊髓组织蛋白以固相pH梯度等电聚焦(immobilized pH gradients isoelectric focusing,IPGIEF)为第一向,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electroporesis,SDS-PAGE)为第二向进行2-DE,应用PDQuest分析软件对考马斯亮蓝染色的2-DE图谱进行图像分析,对差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱(matrixassisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)分析,并利用Mascot查询软件搜索SwissProt数据库进行生物信息学分析。采用蛋白质印迹法验证部分差异蛋白。结果:MT组和NS组大鼠脊髓组织2-DE图谱中各分离出约1 000个清晰的蛋白质点,其中明显差异表达的蛋白有36个。采用MALDI-TOF-MS分析,并利用Mascot查询软件搜索Swiss-Prot数据库,共搜索到14个蛋白质,与NS组比较,MT组中表达下调的蛋白质点有2个,表达上调的蛋白点有12个。采用蛋白质印迹法对其中的NADH脱氢酶及伽玛烯醇化酶蛋白质点进行验证,结果与蛋白质组学结果一致。结论:初步建立了吗啡耐受大鼠脊髓组织蛋白质组双向电泳图谱,证明吗啡耐受可以引起脊髓中多种蛋白质的表达改变。展开更多
基金supported by the National Natural Science Foundation (81601728,31500726)the Natural Science Foundation of Hunan Province (2021JJ41002),China。
文摘Objective:Inflammation in the central nervous system plays a crucial role in the occurrence and development of sepsis-associated encephalopathy.This study aims to explore the effects of maresin 1(MaR1),an anti-inflammatory and pro-resolving lipid mediator,on sepsis-induced neuroinflammation and cognitive impairment.Methods:Mice were randomly assigned to 4 groups:A sham group(sham operation+vehicle),a cecal ligation and puncture(CLP)group(CLP operation+vehicle),a MaR1-LD group(CLP operation+1 ng MaR1),and a MaR1-HD group(CLP operation+10 ng MaR1).MaR1 or vehicle was intraperitoneally administered starting 1 h before CLP operation,then every other day for 7 days.Survival rates were monitored,and serum inflammatory cytokines[tumor necrosis factor alpha(TNF-α),interleukin(IL)-1β,and IL-6]were measured 24 h after operation using enzyme-linked immunosorbent assay(ELISA).Cognitive function was assessed 7 days after operation using the Morris water maze(MWM)test and novel object recognition(NOR)task.The mRNA expression of TNF-α,IL-1β,IL-6,inducible nitric oxide synthase(iNOS),IL-4,IL-10,and arginase 1(Arg1)in cortical and hippocampal tissues was determined by real-time reverse transcription PCR(RT-PCR).Western blotting was used to determine the protein expression of iNOS,Arg1,signal transducer and activator of transcription 6(STAT6),peroxisome proliferator-activated receptor gamma(PPARγ),and phosphorylated STAT6(p-STAT6)in hippocampal tissue.Microglia activation was visualized via immunofluorescence.Mice were also treated with the PPARγantagonist GW9662 to confirm the involvement of this pathway in MaR1’s effects.Results:CLP increased serum levels of TNF-α,IL-1β,and IL-6,and reduced body weight and survival rates(all P<0.05).Both 1 ng and 10 ng doses of MaR1 significantly reduced serum TNF-α,IL-1β,and IL-6 levels,improved body weight,and increased survival rates(all P<0.05).No significant difference in efficacy was observed between the 2 doses(all P>0.05).MWM test and NOR task indicated that CLP impaired spatial learning,which MaR1 mitigated.However,GW9662 partially reversed MaR1’s protective effects.Real-time RTPCR results demonstrated that,compared to the sham group,mRNA expression of TNF-α,IL-1β,and iNOS significantly increased in hippocampal tissues following CLP(all P<0.05),while IL-4,IL-10,and Arg1 showed a slight decrease,though the differences were not statistically significant(all P>0.05).Compared to the CLP group,both 1 ng and 10 ng MaR1 decreased TNF-α,IL-1β,and iNOS mRNA expression in hippocampal tissues and increased IL-4,IL-10,and Arg1 mRNA expression(all P<0.05).Immunofluorescence results indicated a significant increase in Iba1-positive microglia in the hippocampus after CLP compared to the sham group(P<0.05).Administration of 1 ng and 10 ng MaR1 reduced the percentage area of Iba1-positive cells in the hippocampus compared to the CLP group(both P<0.05).Western blotting results showed that,compared to the CLP group,both 1 ng and 10 ng MaR1 down-regulated the iNOS expression,while up-regulated the expression of Arg1,PPARγ,and p-STAT6(all P<0.05).However,the inclusion of GW9662 counteracted the MaR1-induced upregulation of Arg1 and PPARγcompared to the MaR1-LD group(all P<0.05).Conclusion:MaR1 inhibits the classical activation of hippocampal microglia,promotes alternative activation,reduces sepsis-induced neuroinflammation,and improves cognitive decline.
文摘目的采用Meta分析的方法评价右美托咪定联合局麻药腹腔滴注用于腹腔镜手术术后镇痛的有效性和安全性。方法检索Cochrane、Embase、Web of Science、Pubmed、CBM、知网、万方和维普等数据库,纳入比较右美托咪定联合局麻药(研究组)与单独局麻药(对照组)腹腔滴注的随机对照试验(RCT)。两名研究者独立进行文献筛选、质量评价、数据提取后,分别用RevMan 5.3和TSAv0.9Beta进行Meta分析和试验序贯分析(TSA)。结果共纳入5项RCT研究,共320例患者。与对照组比较,研究组术后24 h内各时点VAS疼痛评分明显降低(P<0.05),术后镇痛时间明显延长(P=0.002),镇痛药使用量明显减少(P<0.001),术后肩痛发生率明显降低(P<0.001),术后呕吐的发生率明显降低(P<0.05)。两组术后恶心、低血压和心动过缓的发生率差异无统计学意义。结论在腹腔镜手术中右美托咪定作为局麻药的佐剂腹腔滴注增强局麻药的作用效果,但鉴于该给药途径超说明书使用,腹腔内使用时应谨慎。
文摘目的:建立吗啡耐受大鼠腰段脊髓组织蛋白质组双向凝胶电泳(two-dimensional gel electrophoresis,2-DE)图谱,比较分析吗啡耐受大鼠腰段脊髓组织中蛋白质组的变化和差异表达,鉴定出差异表达的蛋白质点并进行验证。方法:将16只鞘内置管雄性SD大鼠随机分为吗啡耐受组(MT组,n=8)和生理盐水组(NS组,n=8),取其腰段脊髓组织蛋白以固相pH梯度等电聚焦(immobilized pH gradients isoelectric focusing,IPGIEF)为第一向,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electroporesis,SDS-PAGE)为第二向进行2-DE,应用PDQuest分析软件对考马斯亮蓝染色的2-DE图谱进行图像分析,对差异蛋白质点进行基质辅助激光解吸电离飞行时间质谱(matrixassisted laser desorption/ionization time of flight mass spectrometry,MALDI-TOF-MS)分析,并利用Mascot查询软件搜索SwissProt数据库进行生物信息学分析。采用蛋白质印迹法验证部分差异蛋白。结果:MT组和NS组大鼠脊髓组织2-DE图谱中各分离出约1 000个清晰的蛋白质点,其中明显差异表达的蛋白有36个。采用MALDI-TOF-MS分析,并利用Mascot查询软件搜索Swiss-Prot数据库,共搜索到14个蛋白质,与NS组比较,MT组中表达下调的蛋白质点有2个,表达上调的蛋白点有12个。采用蛋白质印迹法对其中的NADH脱氢酶及伽玛烯醇化酶蛋白质点进行验证,结果与蛋白质组学结果一致。结论:初步建立了吗啡耐受大鼠脊髓组织蛋白质组双向电泳图谱,证明吗啡耐受可以引起脊髓中多种蛋白质的表达改变。
