氯喹通过抑制NF-κB和MAPK信号通路减轻脂多糖诱导的BV2小胶质细胞炎症反应

目的:探究氯喹(CQ)对脂多糖(LPS)刺激的BV2小胶质细胞活化的抑制作用及可能机制。方法:将小鼠BV2小胶质细胞分为对照组、LPS组和LPS+CQ组。LPS+CQ组预先给予CQ(10μmol/L)处理30 min再给予LPS刺激,在LPS组和LPS+CQ组中给予LPS(500μg/L)刺激后,各组细胞分别培养30 min、6 h和24 h。倒置显微镜下观察BV2细胞的形态学改变;RT-qPCR和ELISA法检测白细胞介素6(IL-6)和肿瘤坏死因子α(TNF-α)的mRNA和蛋白表达水平来评估BV2细胞的活化情况;用免疫荧光染色检测NF-κB蛋白核转移情况;用Western blot检测核因子κB抑制蛋白α(IκB-α)蛋白的表达情况及c-Jun氨基末端激酶(JNK)和p38蛋白的磷酸化水平。结果:LPS刺激后,BV2细胞形态由圆形或椭圆形向多极或纺锤样转变,而预先给予CQ能抑制BV2细胞的形态转变。LPS刺激后,BV2细胞中TNF-α和IL-6的mRNA水平和培养上清液中的蛋白水平明显增加,但预先给予CQ则明显抑制TNF-α和IL-6的mRNA和蛋白表达,可见CQ能减轻LPS诱导的BV2细胞炎症反应。此外,LPS刺激后,BV2细胞内IκB-α蛋白大量降解,细胞核内的NF-κB蛋白显著增多,MAPK信号通路中JNK和p38蛋白磷酸化水平明显升高;而预先给予CQ可显著减少IκB-α蛋白的降解,明显抑制NF-κB蛋白向细胞核内转移,显著降低JNK和p38蛋白的磷酸化水平,可见CQ能抑制LPS诱导下BV2细胞内NF-κB和MAPK信号通路的激活。结论:氯喹显著减轻LPS诱导的BV2细胞炎症反应,其机制与抑制NF-κB和MAPK信号通路有关。     

c-Jun氨基末端激酶与caspase在细胞凋亡中的作用及相互关系

丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)是信号从细胞表面转导到细胞核内部的重要传递者.在真核细胞中,已确定出4条MAPK信号转导通路[1],即细胞外调节蛋白激酶(extracellular regulated protein kinases,ERK) 通路、c-Jun 氨基末端激酶(c-Jun N-terminal kinase,JNK)通路、P38通路及ERK5通路.JNK也称为应激蛋白激酶,参与应激反应和细胞死亡[2].     

微小RNA-155对小胶质BV-2细胞炎症因子分泌及吲哚胺2,3-双加氧酶表达的影响

目的:探讨微小RNA-155(miR-155)对小胶质BV-2细胞炎症因子分泌及吲哚胺2,3-双加氧酶(IDO)表达的影响。方法:采用携带miR-155的慢病毒感染小胶质BV-2细胞,以脂多糖(LPS)处理BV-2细胞为对照。观察细胞形态,流式液相芯片检测炎症因子的分泌水平,real-time PCR检测炎症因子和IDO的mRNA表达水平,Western blot法检测细胞因子信号抑制物1(SOCS1)、磷酸化p38 MAPK和IDO蛋白的蛋白水平。结果:携带miR-155的慢病毒成功感染BV-2细胞,其miR-155表达水平高于LPS处理组和阴性病毒感染组(P0.01)。miR-155促进BV-2细胞白细胞介素6(IL-6)、肿瘤坏死因子α(TNF-α)、单核细胞趋化蛋白1(MCP-1)和IL-10分泌,抑制IL-12分泌,上调IL-6、TNF-α、IL-10和IDO的mRNA表达,同时升高IDO蛋白表达水平和p38 MAPK蛋白磷酸化水平,下调SOCS1蛋白表达(P0.01)。LPS刺激BV-2细胞分泌炎症因子IL-6、TNF-α、MCP-1和IL-12,上调IL-6、TNF-α和IDO mRNA表达,同时上调IDO、p-p38 MAPK和SOCS1的蛋白水平。结论:miR-155促进小胶质BV-2细胞相关炎症因子分泌和IDO蛋白表达,可能与SOCS1和p38 MAPK信号通路相关。     

肿瘤坏死因子α诱导人髓核细胞凋亡的作用途径

背景:在与细胞凋亡有关的众多因素中,肿瘤坏死因子(tumor necrosis factor,TNF)超家族成员发挥了重要的作用。但TNF是通过何种途径诱导椎间盘髓核细胞凋亡的机制尚未阐明。目的:探讨TNF-α激活后,丝裂原活化蛋白激酶(mitogen-activated protein kinases,MAPKs)信号转导通路中P38MAPK和应激活化蛋白激酶/c-Jun氨基末端激酶(stress-activated protein kinase/c-Jun NH2-terminal kinase,JNK/SAPK)两条途径对人髓核细胞凋亡的作用。方法:体外培养人髓核细胞,将细胞随机分成4组:TNF-α刺激组,P38MAPK阻断组,P-JNK/SAPK阻断组和对照组。采用TUNEL法检测髓核细胞凋亡情况,免疫荧光法检测P-P38MAPK和P-JNK/SAPK的表达及定位;Western Blot法检测P38MAPK,JNK/SAPK及其磷酸化形式的表达。结果与结论:TUNEL法检测凋亡结果中,TNF-α刺激组较其他各组的凋亡细胞密度大(P0.01);免疫荧光结果显示TNF-α刺激组P-P38MAPK和JNK/SAPK在细胞质和细胞核的表达高于各阻断组和对照组(P0.01);Western Blot结果显示P38MAPK,P-JNK/SAPK在各组髓核细胞内均有表达,但无活化形式,TNF-α刺激组可见P-P38MAPK,P-JNK/SAPK表达,但相应阻断组无表达。结果表明,外源性TNF-α可通过P38MAPK和P-JNK/SAPK途径导致人髓核细胞凋亡。     

肺炎链球菌表面蛋白C诱导人嗜中性粒细胞分泌IL-8的机制研究

目的研究肺炎链球菌表面暴露的毒性表面蛋白C(PspC)促进IL-8分泌的可能机制。方法使用炎症相关的信号分子NF-κB,p38MARK,ERK,JNK的抑制剂预处理人嗜中性粒细胞后,观察PspC对IL-8分泌的影响。从受PspC刺激的人嗜中性粒细胞中分别提取细胞总蛋白和细胞核提取物,用ELISA法测定p38MAPK蛋白的活力和NF-κB的浓度,并且采用Western blot方法验证PspC对p38MAPK和IκB-α蛋白磷酸化水平的影响。结果使用NF-κB及p38MARK的抑制剂预处理人嗜中性粒细胞后,可以扭转PspC促中性粒细胞分泌IL-8的现象;而ERK和JNK的抑制剂无此效果。同时,PspC可以上调中性粒细胞中的P38MAPK蛋白的含量和NF-κB的浓度,也可以提高p38MAPK和NF-κB通路的抑制蛋白IκB-α蛋白磷酸化水平。结论肺炎链球菌PspC诱导人体中性粒细胞释放IL-8受p38MAPK和NF-κB途径的调控。     

PPARγ配体4i通过抑制NF-κB和MAPK信号通路抑制小鼠巨噬细胞炎性细胞因子的产生

目的探讨新型过氧化物酶体增殖物激活受体γ(PPARγ)配体4i的体外抗炎作用及机制。方法取对数生长期RAW264. 7小鼠腹腔巨噬细胞,经100 ng/m L脂多糖(LPS)诱导,采用ELISA检测10μmol/L 4i对巨噬细胞肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)分泌的影响;采用Western blot法检测4i对核因子κBp65(NF-κBp65)、核因子κB抑制蛋白α(IκBα)、c-Jun氨基端激酶(JNK)、胞外信号调节激酶1/2(ERK1/2)、p38丝裂原激活蛋白激酶(p38MAPK)活性的影响,加入不可逆的PPARγ拮抗剂GW9662(5μmol/L)探讨PPARγ在NF-κB和MAPK相关蛋白表达中的作用;采用SYBYL 8. 1软件进行分子对接分析探讨4i与PPARγ蛋白的结合特性。结果 4i显著抑制TNF-α和IL-6的产生呈时间依赖性;不同程度抑制NF-κBp65、IκBα、JNK、ERK1/2和p38MAPK蛋白的磷酸化水平,且抑制作用被GW9662所逆转; 4i能与PPARγ受体较好地结合。结论4i通过激活PPARγ抑制NF-κB和MAPK信号通路相关蛋白的活化,抑制TNF-α、IL-6等的产生,发挥抗炎作用。     

人参皂苷Rg1对脓毒症所致心肌损伤大鼠NF-κB磷酸化水平及炎症相关通路的影响北大核心CSCD

目的：探究人参皂苷Rg1对脓毒症所致心肌损伤大鼠核因子-κB(NF-κB)磷酸化水平及炎症相关通路的影响。方法：盲肠结扎穿孔法（CLP）构建大鼠脓毒症心肌损伤模型,60只大鼠随机分为假手术（sham）组、CLP组、人参皂苷Rg1低剂量（CLP+Rg1L）组、人参皂苷Rg1中剂量（CLP+Rg1M）组、人参皂苷Rg1高剂量（CLP+Rg1H）组、瑞沙托维（CLP+TAK-242）组,每组10只。LPS构建体外脓毒症心肌细胞模型,H9C2细胞分为对照（control）组、LPS组、LPS+Rg1L组、LPS+Rg1M组、LPS+Rg1H组、LPS+TAK-242组。右颈总动脉插管法检测各组大鼠平均动脉压（MABP）、心率×左心室发展压（LVDP×HR）、左心室压力最大上升/下降速率（±dp/dtmax）水平;HE染色观察心肌组织病理学改变;Annexin V-FITC/PI双染法检测心肌组织和H9C2细胞凋亡;ELISA检测大鼠血清和H9C2细胞肌酸激酶（CK）、乳酸脱氢酶（LDH）、天冬氨酸转氨酶（AST）、肌钙蛋白Ⅰ（cTnⅠ）、肿瘤坏死因子-α(TNF-α)、IL-6、IL-1β含量;CCK-8及EdU染色检测H9C2细胞增殖能力;RT-qPCR检测心肌组织Toll样受体4(TLR4)、NF-κB p65、p38丝裂原活化蛋白激酶（p38MAPK） mRNA表达;Western blot检测心肌组织TLR4、NF-κB p65、p-NF-κB p65、p38MAPK、p-p38MAPK、核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)蛋白表达。结果：人参皂苷Rg1能够提高MABP、LVDP×HR、±dp/dtmax水平,降低CK、LDH、AST活力,降低cTnⅠ、TNF-α、IL-6、IL-1β、TLR4、NF-κB p65、p38MAPK mRNA与TLR4、p-NF-κB p65/NF-κB p65、p-p38MAPK/p38MAPK、NLRP3蛋白水平（P<0.05）,促进心肌细胞增殖,抑制细胞凋亡与炎症水平,改善心肌损伤。结论：人参皂苷Rg1能够抑制心肌组织细胞凋亡及炎症水平,提高心功能,从而减轻脓毒症所致心肌损伤,其机制可能与TLR4/NF-κB、p38MAPK信号通路有关。     

c-Jun氨基末端激酶在脑缺血再灌注损伤中的作用

在生物体内,丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)是信号从细胞表面转导至细胞核内部的重要传递者,是一类丝/苏氨酸残基的蛋白激酶。目前在真核生物细胞中,已明确了4条MAPK信号转导通路,即细胞外信号调节蛋白激酶(extracellular signal-regulated protein kinases,ERK)通路、c-Jun氨基末端激酶(c-Jun N-terminal kinase,JNK)通路、p38通路及ERK5通路。JNK信号通路作为MAPK信号通路中重要的通路之一,参与了多种生理、病理过程,目前诸多研究表明其在脑缺血/再灌注损伤过程中尤其是程序性细胞死亡过程中起着重要的调控作用,本文就近年来对JNK通路的研究及JNK在脑缺血再灌注诱导的细胞凋亡中的作用及机制作一综述。     

麝香保心丸通过抑制p38 MAPK和NF-κB信号传导通路表达对抗高糖诱导的心肌细胞损伤

目的探讨麝香保心丸(SBP)是否通过抑制p38丝裂原活化蛋白激酶(mitogen-activated protein kinase,MAPK)和核因子kappa B(nuclear factor kappa B,NF-κB)通路保护H9c2心肌细胞对抗高浓度葡萄糖(高糖,HG)引起的损伤。方法应用35 mmol/L的HG处理H9c2心肌细胞24 h,建立HG诱导的心肌细胞损伤模型;细胞计数试剂盒8测定细胞存活率;Hoechst 33258核染色荧光显微镜照相法测定细胞凋亡;双氯荧光素(2’,7’-dichlorfluorescein-diacetate,DCFH-DA)染色/荧光显微镜照相法测定胞内活性氧(ROS)水平;罗丹明123(Rh123)染色法测定线粒体膜电位(MMP);Western blot法测定p38MAPK和NF-κB p65蛋白表达水平。结果 HG处理H9c2心肌细胞24 h能引起细胞明显的损伤,使细胞存活率降低,凋亡细胞数量和细胞内ROS生成增多,MMP丢失;HG能增加p38 MAPK和NF-κB p65磷酸化水平;SBP预处理能明显抑制HG上调p38 MAPK和NF-κB p65磷酸化水平这一作用;SBP、p38MAPK通路抑制剂SB203580和NF-κB通路抑制剂PDTC均能阻断HG对心肌细胞的上述损伤作用,包括细胞毒性、凋亡、ROS生成增多及MMP丢失等。结论麝香保心丸(SBP)可通过抑制p38 MAPK和NF-κB信号传导通路保护H9c2心肌细胞对抗高糖(HG)引起的损伤。     

CysLT2受体拮抗剂HAMI3379抑制LPS诱导小鼠小胶质瘤细胞系的炎性反应

目的探究半胱氨酰白三烯2(CysLT2)受体拮抗剂HAMI3379对LPS诱导小鼠小胶质细胞(BV-2)炎性反应的调控作用及其可能的作用机制。方法体外培养BV-2,将BV-2分为对照组、LPS(100 ng/mL)组、HAMI3379(0.01、0.1和1μmol/mL)组和LPS+HAMI3379组。CCK-8法检测BV-2细胞的增殖;ELISA检测细胞上清液中炎性因子IL-1β、TNF-α、IL-10的含量;Western-blot检测PKCα、IKBα、NF-κB p50和p65蛋白的表达。结果LPS能够激活BV-2细胞,促进其细胞的增殖(P<0.05);显著增加细胞上清液中炎性因子IL-1β、TNF-α的分泌,减少IL-10的分泌(P<0.05);且显著上调PKCα、IKBα、p65蛋白的表达水平(P<0.05)。CysLT2受体拮抗剂HAMI3379能够显著减轻上述变化(P<0.05)。结论CysLT2受体拮抗剂HAMI3379能够抑制LPS激活BV-2细胞,抑制炎性反应,其作用机制可能与抑制PKCα/NF-κB信号通路有关。     

三七皂苷Rg1抑制脂多糖诱导的小胶质细胞激活

目的 探讨三七皂苷Rg1(Rg1)对脂多糖(LPS)诱导的小胶质细胞系BV-2细胞炎性因子释放的抑制作用.方法 用LPS刺激BV-2细胞构建炎症模型,采用四甲基偶氮唑盐比色法检测Rg1对BV-2细胞活力的影响,免疫荧光染色和反转录PCR方法检测不同浓度Rg1(10、20、40μmol/L)对细胞炎性蛋白酶诱导型一氧化氮合酶(iNOS)和环氧合酶-2 (COX-2)、细胞炎性因子肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)、炎性信号分子NF-κB蛋白与mRNA的表达变化.结果 不同浓度的Rg1在转录水平和翻译水平上明显抑制了LPS诱导的细胞炎性蛋白酶iNOS和COX-2、细胞炎性因子TNF-α和IL-1β与炎性信号分子NF-κB的上调,并且iNOS、COX-2和NF-κB的表达呈剂量依赖性.结论 Rg1可通过调控LPS诱导的小胶质细胞系BV-2细胞炎性因子释放从而抑制小胶质细胞激活,发挥抗神经炎症的作用.     

Sesamin inhibits lipopolysaccharide-induced cytokine production by suppression of p38 mitogen-activated protein kinase and nuclear factor-kappaB

Sesame seed oil increases the survival after cecal ligation and puncture in mice and the increased IL-10 levels with non-lethal lipopolysaccharides (LPS) challenge. We showed that sesamin and sesamolin, major lignans of sesame oil, regulated LPS-induced nitric oxide production in the murine microglia and BV-2 cell line. In this study, we studied the effect of sesamin on cytokine production by LPS stimulation. The result showed that sesamin significantly inhibited LPS-stimulated IL-6 mRNA and protein, and to a lesser degree TNF-alpha, in BV-2 microglia. Sesamin and sesamolin also reduced LPS-activated p38 mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-kappaB activations. Furthermore, SB203580, a specific inhibitor of p38 MAP kinase, specifically inhibited LPS-induced IL-6 production. These results suggest that sesamin inhibited LPS-induced IL-6 production by suppression of p38 MAPK signal pathway and NF-kappaB activation.     

封闭枯否细胞对LPS诱导激活丝裂原活化蛋白激酶的影响

目的： 利用小鼠LPS血症模型，探讨封闭枯否细胞（KCs）对LPS诱导MAPK信号转导通路的影响。方法： 雄性昆明种小鼠在注射LPS（5 mg/kg）前48 h及24 h，分别静脉注射GdCl₃（10 mg/kg）或等量的生理盐水，于LPS或生理盐水注射后30 min，分别取出肝脏或分离KCs；体外培养小鼠KCs经GdCl₃（100 μmol/L）预处理1 h，加入含LPS（100 μg/L）的DMEM培养基继续孵育30 min，分别检测肝脏及体内外KCs ERK1/2、p38MAPK蛋白表达和磷酸化水平，及GdCl₃对KCs吞噬、分泌功能的影响。结果： GdCl₃可抑制LPS 诱导KCs活化和TNF-α分泌，但不能抑制LPS诱导KCs或肝脏ERK1/2、p38MAPK磷酸化，也不能影响ERK1/2、p38MAPK蛋白表达，KCs 经GdCl₃单独短时间处理（1 h），可使其少量分泌TNF-α。结论： 封闭KCs并不能通过调节细胞内ERK1/2、p38MAPK信号转导途径，抑制LPS诱导的KCs活化及TNF-α释放，而可能是通过其它信号转导途径（JNK、NF-кB、GPCR等）间的相互作用减轻肝损伤。     

Close association of p38 and JNK with plasminogen-dependent upregulation of PAI-1 in rat astrocytes in vitro

As reported previously, stimulation of astrocytes with plasminogen (PLGn) remarkably enhances their production/release of plasminogen activator inhibitor-1 (PAI-1). In addition, both p38 mitogen-activated protein kinase (p38MAPK) and c-Jun N-terminal kinase (JNK) are activated in these astrocytes. However, it remains to be determined whether the MAPK activation is associated with the PAI-1 induction in PLGn-stimulated astrocytes. In the present study, we investigated the relationship between MAPK activity and PAI-1 induction in PLGn-stimulated astrocytes. PLGn stimulation led to definitive phosphorylation of three MAPKs: external signal regulated kinase (ERK), JNK and p38. These results suggest that all of these MAPKs, either alone or in combination, are involved in PAI-1 induction. To verify this association, an inhibition experiment was carried out by using inhibitors specific for each MAPK. The results of the immunoblotting analysis indicated that 20 μM SB203580 (the p38 inhibitor) or SP600125 (the JNK inhibitor) suppressed approximately 85% or 40% of PLGn-inducible PAI-1, respectively. Only 20% inhibition was achieved by pretreatment of astrocytes with 20 μM PD98059 (the inhibitor of MEK1/2, an upstream kinase of ERK). In conclusion, p38 and JNK were shown to be the major MAPKs involved in the signaling cascade leading to PAI-1 induction in astrocytes stimulated with PLGn.     

Involvement of MAPKs in ICAM-1 expression in glomerular endothelial cells in diabetic nephropathy

Inflammatory processes are involved in the pathogenesis of diabetic nephropathy. The aim of this study was to clarify the role of mitogen-activated protein kinase (MAPK) pathways for induction of intercellular adhesion molecule-1 (ICAM-1) expression in glomerular endothelial cells under diabetic conditions. We examined the expression of ICAM-1 in the kidneys of experimental diabetic rats. Human glomerular endothelial cells (GE cells) were exposed to normal glucose concentration, high glucose concentration (HG), or high mannitol concentration (HM), and then the expression of the ICAM-1 protein and the phosphorylation of the 3 subfamilies of mitogen-activated protein kinase (MAPK) were determined using Western blot analysis. Next, to evaluate the involvement of MAPKs in HG- or HM-induced ICAM-1 expression, we preincubated GE cells with the inhibitors for ERK, p38 or JNK 1h prior to the application of glucose or mannitol. Expression of ICAM-1 was increased in the glomeruli of diabetic rats. Both HG and HM induced ICAM-1 expression and phosphorylation of ERK1/2, p38 and JNK in GE cells. Expression of ICAM-1 was significantly attenuated by inhibitors of ERK, p38 and JNK. We conclude that activation of ERK1/2, p38 and JNK cascades may be involved in ICAM-1 expression in glomerular endothelial cells under diabetic conditions.     

盐酸戊乙奎醚抑制脂多糖致急性肺损伤大鼠肺组织p38丝裂原活化蛋白激酶的活化

目的:观察盐酸戊乙奎醚(PHC)对脂多糖(LPS)致急性肺损伤(ALI)大鼠肺组织p38丝裂原活化蛋白激酶(p38MAPK)、c-jun氨基末端激酶(JNK)活化的影响。方法:SD大鼠随机分为对照组、LPS模型组（5 mg/kg LPS,iv）和LPS+PHC高、中、低(3.0、1.0和0.3 mg/kg)3个剂量组,每组6只,进行PHC对肺组织p38MAPK、JNK表达的量效性分析;另取大鼠在注入NS后即刻0（对照组）和注射LPS后2 h、4 h、6 h和12 h共5个时点,每时点6只,进行肺组织p38MAPK、JNK表达的时效性分析。蛋白免疫印迹法检测肺组织p38MAPK、JNK的表达。结果:LPS模型组大鼠肺组织磷酸化p38MAPK、JNK的表达显著高于对照组（P＜0.05);PHC高剂量组显著抑制LPS诱导的大鼠肺组织磷酸化p38MAPK表达（P＜0.05);PHC在造模后6 h时最能有效抑制磷酸化p38MAPK上调。与LPS模型组相比,PHC高、中、低剂量组磷酸化JNK的表达均无显著差异(均P＞0.05);造模后不同时点,PHC对磷酸化JNK的表达均无抑制作用。结论:PHC抑制LPS诱导的ALI大鼠肺组织p38MAPK活化,但不能抑制JNK活化,PHC对LPS诱导大鼠ALI的拮抗作用可能与其抑制p38MAPK的活化有关。     

Nardosinone-Type Sesquiterpenes from the Hexane Fraction of <Emphasis Type="Italic">Nardostachys jatamansi</Emphasis> Attenuate NF-κB and MAPK Signaling Pathways in Lipopolysaccharide-Stimulated BV2 Microglial Cells

Four nardosinone-type sesquiterpenes, nardosinone, isonardosinone, kanshone E, and kanshone B, were isolated from the hexane fraction of Nardostachys jatamansi (Valerianaceae) methanol extract. The structures of these compounds were mainly established by analyzing the data obtained from nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS). In this study, we investigated their anti-neuroinflammatory effects in lipopolysaccharide (LPS)-induced BV2 microglial cells. The results showed that nardosinone-type sesquiterpenes inhibited the production of pro-inflammatory mediators, such as nitric oxide (NO) and prostaglandin E₂ (PGE₂) in LPS-induced BV2 microglial cells. These inhibitory effects were correlated with the downregulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Moreover, these sesquiterpenes also attenuated the mRNA expression of pro-inflammatory cytokines including interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α) in LPS-induced BV2 microglial cells. During the evaluation of the signaling pathways involved in these anti-neuroinflammatory effects, western blot analysis and DNA-binding activity assay revealed that the suppression of inflammatory reaction by these sesquiterpenes was mediated by the inactivation of nuclear factor-kappa B (NF-κB) pathway. These sesquiterpenes also suppressed the phosphorylation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) signaling pathways in LPS-stimulated BV2 microglial cells. Taken together, these four nardosinone-type sesquiterpenes inhibited NF-κB- and MAPK-mediated inflammatory pathways, demonstrating their potential role in the treatment of neuroinflammation conditions.     

LPS诱导胰腺癌细胞Panc-1表达B7-H1

目的: 研究胰腺癌细胞株Panc-1在应用脂多糖(LPS)刺激前后B7-H1表达的变化,并探讨其可能的分子机制。方法: LPS刺激以及丝裂原活化蛋白激酶(MAPKs)的特异性抑制剂处理Panc-1细胞前后,应用Western blotting检测MAPKs信号通路中p38丝裂原活化蛋白激酶(p38)、细胞外信号调节蛋白激酶(ERK)和c-Jun氨基端激酶(JNK)磷酸化水平的变化,real-time PCR和Western blotting检测B7-H1 mRNA和蛋白的表达变化。结果: LPS刺激后B7-H1的表达显著上调,p38、ERK和JNK的磷酸化水平也明显上调,加入MAPKs特异性的抑制剂后,LPS诱导的p38、ERK和JNK的磷酸化被抑制,并且在p38和ERK的抑制剂处理后,B7-H1的表达也明显被抑制,而在JNK的抑制剂处理后B7-H1的表达没有显著变化。结论: LPS可以诱导胰腺癌细胞株Panc-1表达B7-H1,并且p38和ERK的活化在LPS诱导的B7-H1的表达过程中起着重要作用。