Notch1信号通过TRAF6参与TLR4介导的NF-κB激活

目的观察Notch1和TLR4信号交叉调控NF-κB激活并探讨其可能的作用机制。方法体外培养RAW264.7细胞,LPS刺激RAW264.7细胞后检测Notch1信号激活以及Notch1信号抑制剂DAPT对TNF-α、IL-6和IL-1β表达的影响,Western blot检测DAPT对IKKα/β磷酸化以及NF-κB p65活化的影响,并检测DAPT对TLR4信号通路IKKα/β上游信号My D88和TRAF6表达的影响。结果 LPS刺激RAW264.7细胞后能显著提高Notch1信号蛋白NICD和目的蛋白Hes1表达,DAPT能明显抑制NICD和Hes1的表达。LPS诱导TNF-α、IL-6和IL-1β表达和释放能被DAPT抑制但被Notch1信号激动剂jagged1增强。DAPT能够抑制LPS介导的TRAF6表达和IKKα/β磷酸化,促进NF-κB p65核转位,但DAPT对LPS诱导My D88的表达没有明显的影响。结论 Notch1和TLR4信号存在交叉对话,Notch1通过TRAF6调控NF-κB活化参与TLR4介导的免疫应答。     

雷帕霉素减轻大鼠脑缺血再灌注损伤与TLR4/MyD88/NF-κB信号通路相关性

目的观察雷帕霉素对大鼠脑缺血再灌注损伤的保护作用及对TLR4/MyD88/NF-κB信号通路的调节。方法 SD大鼠30只,随机分为三组,每组十只,分别为:假手术组(Sham组)、脑缺血再灌注损伤组(I/R组)和雷帕霉素预处理组(RAP组)。HE染色观察脑组织病理改变,TUNEL检测脑组织凋亡,ELISA检测脑损伤标志物和炎症因子的变化,Western blot检测脑组织TLR4、MyD88和NF-κB蛋白表达。结果雷帕霉素改善缺血再灌注造成的脑组织损伤,改善脑组织损伤造成的细胞凋亡,降低缺血再灌注损伤脑组织脑损伤标志物及炎症因子水平,雷帕霉素降低缺血再灌注损伤脑组织TLR4信号通路相关蛋白表达。结论雷帕霉素减轻大鼠脑缺血再灌注损伤与抑制TLR4/MyD88/NF-κB信号通路相关。     

川芎嗪抑制TLR4/MyD88/NF-κB信号通路减轻子宫内膜异位症大鼠炎症反应北大核心CSCD

目的探讨川芎嗪抑制Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子κB(NF-κB)信号通路减轻子宫内膜异位症(EMs)大鼠炎症反应的作用。方法自体子宫移植法构建EMs大鼠模型,随机分为EMs组、川芎嗪(低、中、高)剂量组、阳性药物组,每组各12只;进行假手术的12只大鼠作为假手术组。游标卡尺测定异位病灶体积,HE染色检测异位内膜组织病理学改变,ELISA检测血清性激素及炎症因子水平,qRT-PCR、Western blot、免疫荧光染色分别检测异位内膜组织中炎症因子mRNA表达、TLR4/MyD88/NF-κB信号通路相关蛋白表达及NF-κB p65入核情况。结果与假手术组比较,EMs组大鼠异位内膜组织出现明显病理损伤,血清雌二醇(E2)、孕激素(P)含量、肿瘤坏死因子-α(TNF-α)、白介素(IL)-1β、IL-6水平及异位内膜组织中TNF-α、IL-1β、IL-6 mRNA、总蛋白TLR4、MyD88、p-NF-κB p65和核蛋白NF-κB p65表达水平升高,血清促卵泡激素(FSH)、促黄体生成素(LH)含量降低(P<0.05),同时NF-κB p65蛋白大量入核;与EMs组比较,川芎嗪(低、中、高)剂量组和阳性药物组可缓解EMs模型大鼠的上述改变并降低异位病灶体积,且川芎嗪高剂量组和阳性药物组对EMs大鼠的作用效果较好且相近。结论川芎嗪可能通过抑制TLR4/MyD88/NF-κB信号通路激活减轻EMs大鼠炎症反应。     

Notch通路在大鼠肾脏缺血再灌注损伤TLR4介导的炎症反应中的作用

 目的: 探讨Notch通路对大鼠肾脏缺血再灌注损伤(IRI)中Toll样受体4(TLR4)介导的炎症反应的作用。方法: 雄性SD大鼠75只随机分为假手术组(sham组)、缺血再灌注组(IRI组)和γ-分泌酶抑制剂DAPT干预组(DAPT组)。分别于再灌注6 h、12 h、24 h、48 h、72 h时点观察各组肾脏病理改变,检测血尿素氮(BUN)和血清肌酐(Scr)水平,ELISA检测血清肿瘤坏死因子-α(TNF-α)和白细胞介素-6(IL-6)水平,免疫组化和Western blot分别检测大鼠肾脏Notch1、TLR4和NF-κB p65蛋白表达水平。结果: 在IRI组,呈现不同程度的以肾小管上皮细胞和间质损伤为主的肾脏病理改变,BUN、Scr及血清炎性因子TNF-α、IL-6含量在各时点均显著高于sham组(P<0.05),而在DAPT干预组,在各时点肾脏病理损伤明显减轻,BUN、Scr和血清TNF-α、IL-6水平均显著低于IRI组(P<0.05)。Notch1、TLR4和NF-κB p65主要表达于肾小管上皮细胞胞质中,在sham组仅有微量表达,在IRI组则有高表达,在各时点表达与sham组相比均显著增强(P<0.05),而在DAPT组,各因子的表达水平在各时点均较IRI组显著降低(P<0.05)。结论: 大鼠肾脏IRI出现显著的肾功能及肾脏病理改变,血清炎性因子TNF-α和IL-6水平升高,Notch1、TLR4及NF-κB p65在肾组织中表达增强;而DAPT可通过抑制Notch1活化和TLR4/NF-κB通路,抑制TLR4所介导的炎症反应,从而发挥肾脏保护作用。     

HIF-1α抗心肌缺血再灌注炎性损伤的作用

目的:本文旨在通过稳定低氧诱导因子1α(HIF-1α)表达,探讨其在大鼠心肌缺血再灌注(I/R)所致炎性损伤中的作用及可能的机制。方法:60只清洁级雄性Wistar大鼠随机分为4组:假手术(sham)组、I/R组、HIF-1α稳定剂二甲基乙二酰甘氨酸(DMOG)+I/R组和HIF-1α抑制剂YC-1+I/R组。各组分别予相应处理后,用Western blot法检测心肌组织的Toll样受体4(TLR4)和核因子κB(NF-κB)的蛋白表达;real-time PCR检测白细胞介素1β(IL-1β)、IL-6、肿瘤坏死因子α(TNF-α)、TLR4和NF-κB的mRNA水平;并利用试剂盒检测心肌组织的髓过氧化物酶(MPO)活性;HE染色观察炎性细胞浸润情况。结果:HIF-1α可明显降低心肌组织中炎性细胞的浸润、MPO活性及炎症因子IL-1β、IL-6和TNF-α的mRNA水平,同时降低TLR4和NF-κB的mRNA及蛋白水平(P0.05)。结论:HIF-1α可减轻心肌I/R引起的炎性损伤,其机制可能与抑制TLR4/NF-κB信号通路激活有关。     

长春西汀减轻脑出血大鼠脑组织的炎症损伤

目的:探讨长春西汀对脑出血大鼠的干预作用以及对炎症损伤的影响。方法:将大鼠随机分为假手术组、脑出血组以及长春西汀低剂量组、中剂量组和高剂量组。除假手术组外,其余大鼠均通过注射VII型胶原酶的方法建立脑出血模型,长春西汀低剂量组、中剂量组和高剂量组分别给予腹腔注射0.5、1.0和1.5 mg/kg长春西汀,每天1次,共给药7 d。给药完成后,对各组大鼠进行神经功能缺损评分,称重法计算脑组织的含水量,检测脑组织中髓过氧化物酶(MPO)活性,Western blot法检测脑组织中Toll样受体4(TLR4)、核因子κB(NF-κB)、细胞间黏附分子1(ICAM-1)与血管细胞黏附分子1(VCAM-1)的蛋白表达。结果:与脑出血组相比,给予长春西汀干预的大鼠神经功能缺损评分显著下降(P0.05),脑组织含水量明显下降(P0.05),MPO活性显著降低(P0.05),脑组织中TLR4、NF-κB、ICAM-1与VCAM-1的蛋白表达显著下降(P0.05)。结论:长春西汀对脑出血大鼠脑组织具有保护作用,可能是通过抑制TLR4诱导的NF-κB信号通路以及ICAM-1与VCAM-1的表达来抑制炎症反应。     

Notch1与TLR2相互作用调控CD14~+单核细胞功能在冠状动脉粥样硬化性心脏病中的作用

目的观察Notch1/2分子和Toll样受体(TLR)在冠心病患者中的表达变化,探讨Notch信号通路与TLR2的相互作用对CD14~+单核细胞的调控作用。方法本研究入组稳定性心绞痛(SA)22例、不稳定性心绞痛(UA)34例、急性心肌梗死(AMI)27例,同时入组31例健康对照者。实时定量PCR法检测PBMC中Notch1/2以及TLR1-10 mRNA的相对表达量。分选CD14~+单核细胞,应用Notch信号通路抑制剂DAPT和/或TLR2激动剂Pam3Csk4刺激培养24 h,检测TLR2和Notch信号通路相关分子的表达变化,ELISA法检测上清中促炎细胞因子的表达变化,Western blot法检测细胞中抗髓样分化因子88(MyD88)和TIR结构域接头分子(TRIF)的水平。结果 Notch1 mRNA的相对表达量在AMI组显著升高,Notch2 m RNA的相对表达量在各组间的差异无统计学意义。TLR4、TLR7、TLR9 mRNA的相对表达量在SA组、UA组和AMI组中均显著高于NC组,TLR2 mRNA的相对表达量在AMI组中显著高于NC组、SA组和UA组。DAPT刺激可降低CD14~+单核细胞中TLR2 m RNA的相对表达量,对TLR4、TLR7和TLR9 mRNA的表达无显著影响。Pam3Csk4刺激可升高AMI患者CD14~+单核细胞中Notch1 mRNA的相对表达量,Notch信号通路相关分子Hes1和Hes5 mRNA的相对表达量亦显著升高。DAPT刺激还可抑制AMI患者中TLR2介导的炎症信号通路的应答,降低Pam3Csk4刺激后AMI患者CD14~+单核细胞培养上清中白细胞介素(IL)-6、IL-8和肿瘤坏死因子-α的表达,这一过程主要通过影响TIR结构域接头分子表达及核因子-κB的磷酸化而实现。结论 Notch1和TLR2的相互作用可能发挥调控冠心病患者CD14~+单核细胞功能的作用。     

富氢液对脓毒血症大鼠肠黏膜损伤及TLR4/Myd88/NF-κB信号分子的影响

目的探讨富氢液(HRS)对脓毒血症大鼠肠黏膜损伤及其对TLR4/Myd88/NF-κB信号通路表达的影响。方法 SD大鼠30只,随机分为假手术组(sham组)、脓毒症组(SEP组)和富氢液处理组(C组),每组10只;Sham组仅开腹,暴露盲肠,C组和SEP组行盲肠结扎穿孔术,再分别静脉给予富氢水和等量生理盐水;三组均于模型建立后4h处死大鼠,观察小肠组织病理学结果;ELISA检测血浆中肠脂肪酸结合蛋白(I-FABP)、D-乳酸和炎症因子TNF-α、IL-6、IL-10浓度;Western blot检测肠组织TLR4、Myd88和NF-κB表达。结果富氢液可以减轻脓毒血症大鼠肠黏膜损伤,降低血浆中I-FABP、D-乳酸含量,降低炎症因子TNF-α、IL-6表达,促进IL-10升高(P0.05),抑制肠组织中TLR4、Myd88、NF-κB表达(P0.05)。结论富氢液对脓毒血症大鼠肠黏膜屏障有保护作用,可能与TLR4/Myd88/NF-κB信号通路有关。     

青蒿琥酯对小鼠巨噬细胞RAW264.7中TLR4介导的炎症通路的抑制作用

目的观察青蒿琥酯(artesunate,AS)对脂多糖(lipopolysaccharide,LPS)刺激小鼠RAW264.7细胞Toll样受体4(Toll-like receptor 4,TLR4)介导炎症通路活化的影响,以探讨AS的抗炎作用机制。方法采用免疫荧光观察胞内TLR4表达和分布;免疫印迹检测TLR4及下游炎症通路关键分子的表达和活化;酶联免疫吸附法检测细胞培养上清中TNF-α、IL-6浓度。结果 AS对LPS诱导的TLR4表达及其在细胞中的聚集均有抑制作用;AS同时抑制TLR4衔接蛋白髓分化因子88和含TIR结构域干扰素诱导衔接蛋白表达,也抑制依赖于二者活化的肿瘤坏死因子受体相关因子6表达;对下游MAPK通路,AS抑制p38表达和磷酸化、JNK磷酸化,但对ERK1/2无显著影响;对下游NF-κB通路,AS下调抑制性-κBα(inhibitory-κBα,IκBα)的磷酸化,进而减少NF-κB亚基p50和p65活化入核的数量;最后,AS能够抑制LPS诱导的TNF-α和IL-6释放。结论 AS通过抑制LPS诱导的TLR4通路活化,减少致炎细胞因子的释放,从而发挥其抗炎作用。     

酮咯酸氨丁三醇抑制膝骨关节炎模型大鼠炎性反应

目的探究酮咯酸氨丁三醇(KT)对膝骨关节炎(KOA)模型大鼠炎性疼痛的影响,并从Toll样受体4(TLR4)/核因子-κB(NF-κB)炎性通路初步探究其作用机制。方法取大鼠,用随机数字表法分为:对照组、模型组、KT低(1 mg/kg)、高(4 mg/kg)剂量组、TAK-242组(TLR4拮抗剂,1 mg/kg);除对照组外,其余各组均用改良Hulth法复制大鼠右膝KOA模型;肌肉注射KT 1次/d,经尾静脉注射TAK-2422次/周。观察大鼠膝关节肿胀程度、活动状况,对自发疼痛行为步态评分及检测热痛阈值;取滑膜组织,HE及Masson染色检测组织病理形态变化及纤维组织增生状况;ELISA检测滑膜组织炎性因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)水平;Western blot检测神经突蛋白(neuritin)及通路蛋白TLR4、NF-κB及p-NF-κB、骨桥蛋白(OPN)、整合素金属蛋白酶4(ADAM4)蛋白表达。结果与对照组相比,模型组大鼠关节肿胀、疼痛、滑膜炎性损伤及纤维组织增生等KOA病理症状严重,滑膜IL-1β、TNF-α水平及疼痛指标neuritin表达升高、TLR4/NF-κB p65通路及相关蛋白OPN、ADAM4表达升高(P<0.05)。与模型组相比,KT低、高剂量组及TAK-242组大鼠KOA疼痛、滑膜炎性反应等病理症状缓解,TLR4/NF-κB p65通路及相关蛋白表达降低(P<0.05)。结论KT可缓解KOA模型大鼠滑膜炎性反应及疼痛症状,其缓解作用可能与阻断TLR4/NF-κB通路激活有关。     

Notch信号通路对脑缺血大鼠神经干细胞移植后神经再生的影响

目的 探讨抑制Notch信号通路促进脑缺血大鼠神经干细胞（NSCs）移植后神经再生的机制。方法 采用大脑中动脉阻塞MCAO）方法构建局灶性脑缺血模型,体外培养NSCs,移植入纹状体缺血区。将40只SD大鼠随机分为假手术组、模型组、移植组（移植神经干细胞）、氮-［氮-(3,5-二氟苯乙酰)-L-丙氨酰］-S-苯基甘氨酸丁酯］（DAPT）+移植组。采用HE染色观察各组大鼠神经元损伤程度,利用免疫组织化学、Western blotting检测各组大鼠脑组织中Notch1、Hes1及Hes5的表达情况。结果 与假手术组相比,模型组神经元损伤严重,出现核固缩和核溶解,Notch1、Hes1及Hes5阳性细胞表达明显增多,Notch1、Hes1及Hes5蛋白表达显著上调（P＜0.05）。与模型组相比,移植组和DAPT+移植组神经元损伤均不同程度缓解,Notch1、Hes1及Hes5阳性细胞均有部分表达,各项蛋白表达均有所降低（P＜0.05）;DAPT+移植组神经元损伤明显恢复,较移植组各项蛋白阳性细胞和蛋白表达进一步降低(P＜0.05)。结论 抑制Notch信号通路可促进脑缺血大鼠神经干细胞移植后的神经再生,其机制主要与下调Notch1、Hes1及Hes5的表达有关。     

骨碎补总黄酮干预Notch信号通路影响骨重建过程中成血管-成骨耦联

背景:骨缺损是骨科临床的难题,骨碎补总黄酮对骨缺损治疗有确切疗效,但是其具体机制尚不明确。目的:基于Notch信号通路探讨骨碎补总黄酮在骨重建过程中对血管生成和骨形成的影响。方法:48只SD大鼠随机分为模型组、中药组、中药+DAPT组、DAPT组,每组12只。构建大鼠股骨缺损模型,分别给予中药骨碎补总黄酮和Notch通路阻滞剂DAPT干预,干预12周后通过X射线片、Micro-CT和苏木精-伊红染色观察骨缺损的成骨情况,免疫组化检测骨组织中血管内皮生长因子、CD31、Hes1、骨形态发生蛋白2、Notch1蛋白的表达。结果与结论:①X射线片、Micro-CT和苏木精-伊红染色显示中药组的成骨效果明显优于其他3组;②免疫组化检测显示中药组骨形态发生蛋白2、血管内皮生长因子、CD31、Hes1、Notch1平均吸光度值较模型组明显增多(P<0.05);中药+DAPT组血管内皮生长因子、Hes1、骨形态发生蛋白2的平均吸光度值大于DAPT组(P<0.05),但CD31和Notch1表达组间差异无显著性意义;③结果表明,骨碎补总黄酮通过激活Notch通路促进骨重建过程中成血管-成骨耦联,从而促进骨修复。     

重组人内皮抑素通过激活Dll4/Notch通路抑制大鼠动脉粥样硬化斑块内血管新生

目的:观察重组人内皮抑素(rh ES)对大鼠动脉粥样硬化(AS)斑块内新生血管的抑制作用,探讨Dll4/Notch信号通路在其中的调控机制。方法:雄性Wistar大鼠随机分为正常对照组(N组)、AS组和AS+rh ES组,每组10只。N组始终喂基础饲料,其余2组采用高脂喂养、维生素D3负荷及球囊损伤动脉内膜建立大鼠AS模型。AS+rh ES组以10 mg·kg~(-1)·d~(-1)的rh ES腹腔注射4周,另2组注射等量生理盐水。采血检测各组的总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、C反应蛋白(CRP)、白细胞介素-1(IL-1)和肌钙蛋白I(Tn I)等;免疫组化CD31染色观察新生血管密度;Western blot法检测主动脉内Dll4、Notch1蛋白表达。结果:与N组比较,AS组和AS+rh ES组的TC、TG、LDL-C、CRP和L-1水平均显著升高(P0.05),但上述指标在AS组和AS+rh ES组之间差异无统计学显著性。CD31染色结果显示,AS组的新生血管表达最丰富;与AS组比较,AS+rh ES组的新生血管密度显著下降(P0.05)。AS组的Dll4和Notch1蛋白水平显著低于N组(P0.05);相比AS组,AS+rh ES组的Dll4和Notch1蛋白水平显著升高(P0.05)。结论:rh ES能够抑制大鼠AS斑块内血管新生,Dll4/Notch通路的激活可能是rh ES抑制斑块内的血管新生的信号机制。     

绿茶多酚通过抑制TLR4通路减轻蛛网膜下腔出血大鼠早期脑损伤

目的 探讨绿茶多酚通过抑制TLR4通路对蛛网膜下腔出血大鼠早期脑损伤的影响.方法 建立大鼠蛛网膜下腔出血模型,随机分为模型组、绿茶多酚组、TAK-242(TLR4抑制剂)组、绿茶多酚+TAK-242组,每组12只;另取12只大鼠设为假手术组.药物处理后,对所有大鼠进行神经功能缺损评分,检测各组大鼠脑组织含水量,采用Ev...     

Blockade of Notch signaling promotes acetaminophen-induced liver injury

Liver injury after experimental acetaminophen treatment is mediated both by direct hepatocyte injury through a P450-generated toxic metabolite and indirectly by activated liver Kupffer cells and neutrophils. This study was designed to investigate the role of Notch signaling in the regulation of innate immune responses in acetaminophen (APAP)-induced liver injury. Using a mouse model of APAP-induced liver injury, wild-type (WT) and toll-like receptor 4 knockout (TLR4 KO) mice were injected intraperitoneally with APAP or PBS. Some animals were injected with γ-secretase inhibitor DAPT or DMSO vehicle. For the in vitro study, bone marrow-derived macrophages (BMMs) were transfected with Notch1 siRNA, TLR4 siRNA, and non-specific (NS) siRNA and stimulated with LPS. Indeed, paracetamol/acetaminophen-induced liver damage was worse after Notch blockade with DAPT in wild-type mice, which was accompanied by significantly increased ALT levels, diminished hairy and enhancer of split-1 (Hes1), and phosphorylated Stat3 and Akt but enhanced high mobility group box 1 (HMGB1), TLR4, NF-κB, and NLRP3 activation after APAP challenge. Mice receiving DAPT increased macrophage and neutrophil accumulation and hepatocellular apoptosis. However, TLR4 KO mice that received DAPT reduced APAP-induced liver damage and NF-κB, NLRP3, and cleaved caspase-1 activation. BMMs transfected with Notch1 siRNA reduced Hes1 and phosphorylated Stat3 and Akt but augmented HMGB1, TLR4, NF-κB, and NLRP3. Furthermore, TLR4 siRNA knockdown resulted in decreased NF-κB and NLRP3 and cleaved caspase-1 and IL-1β levels following LPS stimulation. These results demonstrate that Notch signaling regulates innate NLRP3 inflammasome activation through regulation of HMGB1/TLR4/NF-κB activation in APAP-induced liver injury. Our novel findings underscore the critical role of the Notch1-Hes1 signaling cascade in the regulation of innate immunity in APAP-triggered liver inflammation. This might imply a novel therapeutic potential for the drug-induced damage-associated lethal hepatitis.     

Blockade of the Notch1/Jagged1 pathway in Kupffer cells aggravates ischemia-reperfusion injury of orthotopic liver transplantation in mice

Abstract

Liver ischemia-reperfusion injury (IRI) represents a risk factor for early graft dysfunction and an obstacle to expanding donor pool in orthotopic liver transplantation (OLT). Kupffer cells (KCs) are the largest antigen-presenting cell (APC) group and the primary modulators of inflammation in liver tissues. The vital role of Notch1/Jagged1 pathway in mouse OLT model has been reported, however, its potential therapeutic mechanism is unknown. Here, we made use of short hairpin RNA-Jagged1 and AAV-Jagged1 to explore the effects of Notch1/Jagged1 pathway in OLT. In vitro, blockade of Notch1/Jagged1 pathway downregulated the expression of Hairy and enhancer of split-1 (Hes1) gene, which in turn increased the proinflammatory effects of KCs. Moreover, the anti-inflammatory effects of Notch1/Jagged1 pathway were induced by inhibiting Hes1/gene of phosphate and tension/protein kinase B/Toll-like receptor 4/nuclear factor kappa B (Hes1/PTEN/AKT/TLR4/NF-κB) axis in KCs. In vivo, we used a well-established mouse model of OLT to mimic clinical transplantation. Mice were stochastically divided into 6 groups: Sham group (n?=?15); Normal saline (NS) group (n?=?15); Adeno-associated virus–green fluorescent protein (AAV-GFP) group (n?=?15); AAV-Jagged1 group (n?=?15); Clodronate liposome (CL) group (n = 15); CL+AAV-Jagged1 group (n = 15) . After OLT the liver damage in AAV-Jagged1 group were significantly accentuated compared to the AAV-GFP group. While blockade of Jagged1 aftet clearence of KCs by CL would not lead to further liver injuries. Taken together, our study demonstrated that blockade of Notch1/Jagged1 pathway aggravates inflammation induced by lipopolysaccharide (LPS) via Hes1/PTEN/AKT/TLR4/NF-κB in KCs, and the blockade of Notch1/Jagged1 pathway in donor liver increased neutrophil/macrophage infiltration and hepatocellular apoptosis, which suggested the function of Notch1/Jagged1 pathway in mouse OLT and highlighted the protective function of Notch1/Jagged1 pathway in liver transplantation.     

雷公藤多苷联合地塞米松对EAE中TLRs/NF-κB信号通路的作用

 目的：探讨雷公藤多苷联合地塞米松对实验性变态反应性脑脊髓炎（EAE）的治疗效果及其作用途径。方法：所有实验动物被分为空白对照组、EAE组、治疗组1(地塞米松)和治疗组2（雷公藤多苷联合地塞米松）。比较各组平均临床评分;分别采用实时定量RT-PCR和免疫组化法检测不同组别脑组织中TLR4和TLR9 mRNA和蛋白含量,免疫组化法检测NF-κB p65蛋白的表达;采用ELISA法检测外周血中TNF-α、IL-1β和IL-6含量的变化。结果：各治疗组平均临床评分较EAE组均降低（P<0.05）;治疗组2较治疗组1平均临床评分降低（P<0.05）。在发病高峰期,EAE组TLR4和TLR9 mRNA及蛋白表达较空白对照组增高,而各治疗组较EAE组降低（均P<0.05）;EAE组NF-κB p65阳性表达率较各治疗组升高（P<0.05）;各治疗组炎症细胞因子TNF-α、IL-1β和IL-6含量较EAE组降低（P<0.05）。治疗组2与治疗组1比较,上述各指标差异显著（均P<0.05）,且雷公藤多苷与地塞米松具有交互作用（F=75.749,P<0.01）。
结论：TLRs/NF-κB信号通路可能参与了EAE的发病过程;雷公藤多苷联合地塞米松具有明显改善EAE临床症状的效果,其可能通过抑制TLRs/NF-κB信号通路发挥抗炎及免疫抑制双重疗效。     

Involvement of Notch1/Hes signaling pathway in ankylosing spondylitis

We aimed to investigate the role of Notch1/Hes signaling pathway in the pathogenesis of abnormal ossification of hip ligament in patients with ankylosing spondylitis (AS). 22 AS patients scheduled for artificial hip arthroplasty were randomly chosen as AS group. As controls, we used 4 patients diagnosed with transcervical fracture who underwent hip replacement surgery. Notch1 and Hes mRNA expressions were detected by real-time fluorescent quantitative polymerase chain reaction (RFQ-PCR). Immunohistochemistry (IHC) was used to detect Notch1 and Hes protein expression. Correlation analyses of Notch-l and Hes with AS-related clinical factors were conducted with spearman’s correlation analysis and partial correlation analysis. RFQ-PCR results showed significant differences in Notch1 and Hes mRNA expressions between AS group and the control group (all P < 0.05). IHC analysis further indicated positive nuclear signals of Notch1 and Hes protein, indicating functional activation of the Notch1 and Hes pathways. Semi-quantitative IHC showed a higher Notch1 and Hes expression levels in AS group compared to the control group (all P < 0.05). Correlation analysis suggested that Hes protein expression was positively associated with the clinical course of the disease in AS patients. In conclusion, Notch1 and Hes overexpression was clearly detected in hip joint ligaments of AS patients, Hes protein expression was associated with the clinical course of AS. Taken together, we suggest that signaling pathways mediated by Notch1-Hes may contribute to ligament ossification of hip joints in AS patients.