褪黑素减轻大鼠颈动脉球囊损伤后的内膜增生

目的探讨褪黑素能否够减轻大鼠颈动脉球囊损伤后的内膜增生。方法将20只雄性大鼠随机分为对照组、褪黑素组(5 mg/kg)、球囊损伤组(建立颈动脉球囊损伤模型)和褪黑素干预组。14 d后取大鼠的颈动脉,弹力蛋白染色并计算各组大鼠颈动脉的内膜面积与中膜面积的比值(i/m);Western blot检测颈动脉内NF-κB、IL-1β、IL-6和TNF-α的表达。结果球囊损伤能够明显诱导大鼠颈动脉i/m值的增加(P0.01);褪黑素干预组i/m值明显回降(P0.01);球囊损伤组颈动脉的NF-κB、IL-1β、IL-6和TNF-α的表达明显高于对照组及褪黑素组(P0.01);褪黑素干预组NF-κB、IL-1β、IL-6和TNF-α的表达明显回降(P0.05)。结论球囊损伤能够诱导大鼠颈动脉的炎性反应及内膜增生,褪黑素能通过抑制NF-κB介导的炎性反应减轻大鼠颈动脉球囊损伤后的内膜增生。     

缬沙坦对动脉粥样硬化兔核因子κB和单核细胞趋化因子-1的影响

目的:探讨血管紧张素Ⅱ1型受体(AT1R)拮抗剂缬沙坦(Val-sartan)对动脉粥样硬化(AS)兔核因子κB(NF-κB)和单核细胞趋化因子-1(MCP-1)的影响。方法:24只雄性日本大耳白兔随机分为3组:正常对照组,AS模型组,缬沙坦治疗组。喂养12周,进行血脂测定、主动脉内膜/中膜比值测定、主动脉NF-κB和MCP-1的表达和蛋白质含量测定。结果:AS模型组NF-κB和MCP-1蛋白含量显著增加(P<0.05),缬沙坦治疗组显著减少(P<0.05),且NF-κB活化和MCP-1表达之间成正相关(r=0.728,P<0.01);缬沙坦治疗组及AS模型组的血清胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白(LDL)均无差别(P均>0.05),但均高于正常对照组。与AS模型组比较,缬沙坦治疗组主动脉内膜/中膜厚度比值明显减少(P<0.05)。结论:缬沙坦可以干预AS的形成,其机制可能与其抑制NF-κB的活化从而下调MCP-1的表达有关。     

辛伐他汀对高血压大鼠主动脉NF-κB和MCP-1表达的影响

目的研究他汀类药物对核因子κB(NF -κB)、单核细胞趋化蛋白 -1(MCP -1)在自发性高血压大鼠 (SHR)大血管中表达的影响 ,探讨其预防动脉粥样硬化 (AS)的可能机制。方法20只12周龄雄性SHR随机分为SHR组和辛伐他汀 (simvastatin)组,simvastatin组用simvastatin5mg/kg·d灌胃8周 ,另取10只同龄雄性京都种Wistar大鼠为WKY组 ,免疫组化测各组主动脉NF -κB、MCP-1的表达 ,ELISA测血清MCP-1含量。结果SHR组主动脉组织中NF-κB、MCP-1表达及血清MCP-1含量显著增加 (P<0.01) ,且MCP -1表达与NF -κB活化正相关(r=0.728,P<0.01) ;simvastatin组NF -κB、MCP -1表达和血清MCP -1含量显著降低 (P<0.01)。SHR组及simvastatin组之间血压及血清总胆固醇、总甘油三脂水平无明显差异(P>0.05)。结论他汀类药物可能独立于降脂外部分通过抑制NF-κB的活化来调控MCP-1表达而抗AS。     

血府逐瘀汤含药血清对Toll 样受体4 信号转导通路及下游炎症因子的影响

目的:研究血府逐瘀汤含药血清对血管内皮细胞Toll样受体4信号转导通路及下游LOX-1、TNF-α、VCAM-1及ICAM-1等炎症因子表达的影响,探讨血府逐瘀汤抗动脉粥样硬化的机制。方法:采用药物血清学方法制备血府逐瘀汤含药血清以备细胞实验使用。体外培养血管内皮细胞,随机分为对照组、模型组、阿托伐他汀组、血府逐瘀汤组,用LPS刺激2 h后,分别加入阿托伐他汀和血府逐瘀汤含药血清干预24 h。用荧光定量PCR方法测定TLR4、MYD88、TRAF-6、NF-κB、LOX-1、TNF-α、VCAM-1及ICAM-1的基因表达;用Western blot方法测定TLR4、NF-κB、LOX-1、TNF-α、VCAM-1及ICAM-1蛋白表达。结果:用LPS刺激血管内皮细胞后,引起TLR4、My D88、TRAF-6、NF-κB、LOX-1、TNF-α、VCAM-1及ICAM-1的表达升高,与对照组比较,差异有统计学意义(P0.01);用药物血清干预以后血府逐瘀汤组显著抑制各项指标的表达,与模型组比较,差异有统计学意义(P0.05)。结论:血府逐瘀汤可抑制TLR4/NF-κB信号转导通路及下游LOX-1、TNF-α、VCAM-1及ICAM-1等炎症因子的表达,这可能是其防治动脉粥样硬化作用的机制之一。     

基于ERK1/2/核因子κB信号通路探讨大建中汤治疗肠易激综合征内脏痛

目的 探讨大建中汤是否通过干预ERK1/2/核因子κB（NF-κB）通路及其下游分子发挥治疗肠易激综合征（IBS）内脏痛。 方法 采用母婴分离、乙酸灌肠、鸡卵清白蛋白腹腔注射等方法制备IBS内脏痛大鼠模型,大鼠随机分为正常组(生理盐水)、模型组(生理盐水)、大建中汤（10.8g/kg）治疗组和匹维溴铵(45mg/kg)对照组。每组8只,连续灌胃14 d。评估大鼠内脏敏感性采用腹壁撤退反应(AWR);采用Real-time PCR检测各组大鼠结肠组织ERK1/2 mRNA、核因子κB（NF-κB）mRNA、环氧化酶-2 (COX-2 )mRNA及基质金属蛋白酶9 (MMP-9) mRNA的表达;采用免疫组织化学染色法检测各组大鼠结肠组织NF-κB及COX-2的表达。 结果 与正常大鼠相比,模型大鼠于60、40及20 mmHg压力下AWR评分明显升高(P<0.05,P<0.01),ERK1/2、NF-κB、COX-2和MMP-9 表达明显上升(P<0.05);与模型大鼠相比,大建中汤组(60、40和20 mmHg 压力)的 AWR评分明显下降(P<0.05,P<0.01),ERK1/2、 NF-κB、COX-2和MMP-9表达下降明显 (P<0.05);与匹维溴铵对照组相比,大建中汤治疗组ERK1/2、NF-κB、 COX-2和MMP-9 表达差异无统计学意义（P>0.05）。 结论 大建中汤通过干预ERK1/2/NF-κB通路及其下游分子,发挥治疗IBS内脏痛的作用。     

促红细胞生成素抑制大鼠体外循环术致急性肾损伤时NF-κB P65和ICAM-1表达

 目的 研究大鼠体外循环（CPB）术致急性肾损伤时肾脏核转录因子-κB（ NF-κB ）P65和内皮细胞表面黏附分子-1（ICAM-1）表达水平变化及促红细胞生成素(EPO)对其的抑制作用。方法 30只雄性SD大鼠随机分为3组（每组n=10）:Sham组、CPB组和EPO组, Sham组建立CPB模型,不行体外转流,其余2组行体外转流,EPO组于转流前在预充液中加入3 000U/kg的EPO。分别于肝素化后转流前 (T0)和转流结束后(T1)、术后0.5 (T2)、1 (T3)、2 ( T4)和24 h (T5)检测血清肌酐水平, 术后24h取肾脏组织,HE染色观察组织形态,并分别采用免疫组化和western-blot法检测肾组织中NF-κB P65和ICAM-1表达。结果 术后CPB组各时间点与sham组比较血清肌酐水平均明显升高（P<0.05）,与CPB组相比,EPO组的肌酐水平明显下降（P<0.05）;CPB组肾组织中NF-κB p65及ICAM-1表达水平均明显高于sham组（P<0.05）,EPO组NF-κB p65及ICAM-1表达水平均低于CPB组（P<0.05）;病理学检查显示EPO组的肾小管上皮细胞肿胀、胞质内空泡形成、间质出血明显减轻。结论EPO可抑制大鼠体外循环后肾脏NF-κB P65及ICAM-1表达,从而减轻大鼠体外循环引起的肾脏损伤。     

复荣通脉胶囊对糖尿病大鼠血清ADMA水平及胸主动脉eNOS、NF-κB表达的影响

目的:观察复荣通脉胶囊对糖尿病(DM)大鼠血清非对称性二甲基精氨酸(ADMA)水平、内皮型一氧化氮合酶(eNOS)、核转录因子-κB(NF-κB)表达水平的影响,进一步探讨其防治DM大血管病变的作用机制。方法:60只Wistar大鼠先分为空白对照组(n=10)和造模组(n=50),后者复制DM模型后,再分为DM组(n=9)、复荣通脉低剂量组(n=10)、复荣通脉中剂量组(n=9)和复荣通脉高剂量组(n=9),低、中、高剂量组分别给予0.7、1.4、2.8g/kg体重复荣通脉胶囊灌胃,其余两组给予等量纯净水灌胃。8周后,从胸主动脉采血检测ADMA水平;留取胸主动脉制备病理切片,SP法免疫组化染色观察胸主动脉eNOS和NF-κB阳性染色面积及强度。结果:与空白对照组比较,DM组及复荣通脉低、中、高剂量组血清ADMA水平及组织NF-κB表达明显增加(P0.01),组织eNOS表达明显降低(P0.01);与DM组比较,复荣通脉低、中、高剂量组血清ADMA水平及组织NF-κB表达明显降低(P0.01),组织eNOS表达明显升高(P0.01);中、高剂量组ADMA、NF-κB的降低和eNOS的升高更明显(P0.05)。结论:复荣通脉胶囊通过降低DM大鼠血清ADMA水平,上调胸主动脉eNOS和下调NF-κB的表达,对大血管病变发挥保护作用。     

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。     

PDTC对肝纤维化大鼠核转录因子-κB、α-平滑肌肌动蛋白和Ⅰ型胶原表达影响研究

目的探讨吡咯烷二硫代氨基甲酸酯(PDTC)对二甲基亚硝胺(DMN)所致肝纤维化大鼠肝组织中核转录因子-κB(NF-κB)、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原(Col-1)表达的影响及其意义。方法雄性SD大鼠38只,随机分为A、B、C组。A组为正常对照组,B、C组再各自随机分为2周及4周组并腹腔注射DMN诱导大鼠肝纤维化模型,C组造模同时给予PDTC灌胃。应用化学发光凝胶电泳迁移率(EMSA)检测肝组织NF-κBp65活性;采用实时定量聚合酶链反应(Real-time PCR)检测肝组织NF-κBp65和Col-1的mRNA表达;应用免疫组化检测NF-κBp65、α-SMA和Col-1的表达。结果 B组及C组NF-κBp65活性及其mRNA表达、Col-1 mRNA表达、NF-κBp65免疫组化阳性细胞数、α-SMA及Col-1阳性面积均明显高于A组(P<0.01或P<0.05),且随着时间的延长,4周组高于2周组(P<0.01或P<0.05);同期比较发现C组NF-κBp65活性及其mRNA表达、Col-1 mRNA表达、NF-κBp65免疫组化阳性细胞数、α-SMA及Col-1阳性面积均明显低于B组(P<0.01或P<0.05)。NF-κBp65活性与NF-κBp65 mRNA表达、Col-1 mRNA表达、NF-κBp65免疫组化阳性细胞数、α-SMA及Col-1阳性面积呈正相关(r=0.747,0.780,0.779,0.658,0.780,P均<0.01)。结论 NF-κB与大鼠肝纤维化密切相关;PDTC可能通过抑制NF-κB的活性及α-SMA、Col-1表达从而产生抗肝纤维化作用。     

BDNF对大鼠急性脊髓损伤后NF-κB表达的影响

目的研究脑源性神经营养因子(BDNF)对急性脊髓损伤大鼠核转录因子-κB(nuclear factor-kappa B,NF-κB)表达的变化。方法参照Nystrom压迫方法制作大鼠脊髓压迫损伤模型,成年健康Wistar大鼠72只,雌雄不限,按随机数字表法分为正常对照组8只、损伤组32只、BDNF组32只。免疫组化和Western blot检测各组大鼠NF-κB的表达。结果免疫组化结果显示:损伤组与BDNF组NF-κB阳性产物的平均光密度值(mean optic density,MOD)显著高于正常对照组(P<0.01);而BDNF组与损伤组相比A值明显降低(P<0.01);Western blot结果显示:与正常对照组比较,损伤组与BDNF组NF-κB的灰度值(integrated density value,IDV)与内参照IDV的比值明显升高(P<0.01),而BDNF组则明显低于损伤组(P<0.01)。结论 BDNF很可通过下调NF-κB的表达参与脊髓继发性损伤。     

The Expression of Inflammatory Cytokines on the Aorta Endothelia Are Up-regulated in Pinealectomized Rats

This study was designed to investigate the effect of melatonin on the expression of aortic inflammatory cytokines and its underlying mechanisms in rats. Melatonin deficiency rats (Px, N?=?16) were created by pinealectomy and were fed with normal diet for 16 weeks after the surgery, and compared with sham-operated rats (Con, N?=?14). Serum lipid profile, glucose metabolism parameters, serum oxidative stress and inflammatory biomarkers were evaluated. The expression of inflammatory cytokines in the aorta endothelia was analyzed. To evaluate the signal transduction pathways of melatonin on the expression of cytokines, rat aortic endothelial cell lines (RAECs) were treated with melatonin, and their protein expressions of inflammatory cytokines and phosphorylation levels of relevant signal pathways were detected. At the 16th week after surgery in Px rats, their serum triglyceride, very low density lipoprotein cholesterol, free fatty acid and glucose levels were prominently elevated (all P?<?0.05); serum oxidative stress biomarker malondialdehyde, serum inflammatory biomarkers oxidized low-density lipoprotein, tumor necrosis factor-α, interleukin-6 and C reactive protein were also significantly increased. Meanwhile, the expression of inflammatory cytokines: monocyte chemotactic protein-1 (MCP-1), vascular adhesion molecule 1 (VCAM-1) and matrix metalloproteinase-9 (MMP-9) of the aorta endothelia in Px rats were significantly up-regulated (all P?<?0.05). In vitro, melatonin significantly decreased the expression of MCP-1, VCAM-1 and MMP-9 proteins, along with the suppression of phosphorylation levels of nuclear factor κB (NF-κB)/P65 and p38 mitogen-activated protein kinase (P38-MAPK) in RAECs. Melatonin deficiency elevates the serum inflammatory biomarkers and increases aortic inflammatory responses. Melatonin regulates these inflammatory responses by NF-κB and P38-MAPK involved pathways.     

NF-κB及ICAM-1在脑出血大鼠及过氧化氢损伤大鼠脑微血管内皮细胞中的表达

目的： 探讨脑出血后炎症反应机制及核因子-κB(NF-κB)与细胞间粘附分子-1(ICAM-1)间表达的关系。方法： 建立大鼠脑出血模型及过氧化氢损伤大鼠脑微血管内皮细胞的模型，分别采用免疫组化、原位杂交、免疫细胞化学及Western blotting方法观察NF-κB和ICAM-1的表达。结果： 大鼠脑出血后NF-κB 及ICAM-1表达均增强，ICAM-1的表达高峰（1 d）先于NF-κB（4 d），但在体外实验中，过氧化氢损伤后即刻大鼠脑微血管内皮细胞中NF-κB表达即增加，2 h后ICAM-1表达也上调，NF-κB的抑制剂吡咯烷二硫氨基甲酸（PDTC)可下调NF-κB及ICAM-1表达。结论: 在活性氧损伤中NF-κB作为ICAM-1的活化因子，可上调ICAM-1表达，但在脑出血这一复杂的病理生理变化中，尚有其它因素参与对NF-κB 及ICAM-1表达的调控。     

辛伐他汀对同型半胱氨酸诱导的HUVEC的毒性和炎症反应的影响及分子机制

目的：探讨辛伐他汀对同型半胱氨酸(HCY)诱导的内皮细胞毒性和炎症的抑制作用及其分子机制。 方法: 采用MTT检测细胞活性，通过DCFH-DA检测活性氧的生成，Western blotting分析相关蛋白的表达，凝胶滞留法(EMSA)分析NF-κB的DNA结合活性。 结果： HCY处理后，MTT检测发现内皮细胞存活率显著低于对照，流式细胞仪的结果表明活性氧的水平也显著升高。经不同浓度的辛伐他汀预处理后，可以显著地抑制HCY诱导的内皮细胞存活率的下降以及活性氧水平的升高。Western blotting与ELISA结果发现辛伐他汀抑制TNF-α、IL-6、MCP-1及ICAM-1的表达水平。EMSA和Western blotting检测结果表明辛伐他汀抑制HCY介导的p38磷酸化水平以及通过抑制IκBα磷酸化而抑制IκBα蛋白的降解，阻断HCY诱导NF-κB的激活。 结论： 辛伐他汀对HCY介导的内皮细胞毒性及炎症反应有明显的抑制作用，可能是通过阻断ROS-p38-NF-κB通路实现的。     

转录因子NF-κB在内毒素休克中的作用

目的探讨内毒素休克大鼠组织炎性介质的表达特征及其和核转录因子NFκB(nuclearfactorkappaB)的关系。方法应用免疫组织化学技术检测脂多糖(lippolysaccharice,LPS)内毒素休克大鼠肝肾组织转录因子NFκB、炎性介质ICAM1、VCAM1、iNOS的表达。结果LPS内毒素休克大鼠肝肾组织转录因子NFκBp65,炎性介质ICAM1、VCAM1、iNOS阳性细胞率高于正常对照组;炎性介质ICAM-1、VCAM-1、iNOS阳性细胞率与NF-κBp65阳性细胞率成正相关。用吡咯烷二硫氨基甲酸(pyrrolidinedithiocarbmate,PDTC)抑制转录因子NFκB的内毒素休克大鼠炎性介质ICAM1、VCAM1、iNOS阳性细胞率低于LPS组。结论核转录因子NFκB在LPS引起的大鼠内毒素休克炎性介质的表达中起调节作用。     

Effect of chlorogenic acid on LPS-induced proinflammatory signaling in hepatic stellate cells

Objective and design

This study was aimed at investigating the effect of chlorogenic acid (CGA) on lipopolysaccharide (LPS)-induced proinflammatory signaling in hepatic stellate cells (HSCs).

Methods

An immortalized rat HSC line was cultured in vitro and treated with LPS in the absence or presence of CGA. Reactive oxygen species (ROS) production in the HSCs was monitored by flow cytometer using DCFH-DA. The protein expression levels of toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), nuclear factor-κB (NF-κB), and p-IκB-α were determined by Western blot. The mRNA expression levels of TLR4, MyD88, monocyte chemotactic protein 1(MCP-1), and interleukin 6 (IL-6) were detected by RT-PCR. The levels of MCP-1 and IL-6 in the culture supernatant of HSCs were measured by ELISA.

Results

CGA had no effect on expression of TLR4 and MyD88. However, the treatment of CGA can inhibit LPS-induced production of ROS in HSCs. Meanwhile, CGA can inhibit LPS-induced nuclear translocation of NF-κB and IκB-α phosphorylation in HSCs, as well as NAC (a ROS scavenger). The mRNA expression and the levels of MCP-1 and IL-6 in the culture supernatant of the HSCs in this study were elevated by LPS stimulation and inhibited by CGA treatment, as well as NAC and PDTC (a NF-κB inhibitor).

Conclusion

Our results indicate that CGA can efficiently inhibit LPS-induced proinflammatory responses in HSCs and the anti-inflammatory effect may be due to the inhibition of LPS/ROS/NF-κB signaling pathway.     

Stimulation with thromboxane A2 (TXA2) receptor agonist enhances ICAM-1, VCAM-1 or ELAM-1 expression by human vascular endothelial cells

A previous study reported that intercellular adhesion molecule-1 (ICAM-1) expression by human vascular endothelial cells (HUVEC) is augmented by intracellular signal transmission mainly through the protein kinase C (PKC) system stimulated by TXA₂ receptors. In the present study, we show that a TXA₂ receptor agonist, U46619, augments the expression of not only ICAM-1, but also vascular cell adhesion molecule-1 (VCAM-1) or endothelial leucocyte adhesion molecule-1 (ELAM-1) in HUVEC both at protein and mRNA levels. Pretreatment with SQ29,548 (a TXA₂ receptor antagonist) or PKC inhibitors greatly diminished the extent of U46619-induced mRNA accumulation and surface expression of the adhesion molecules. An inhibitor of nuclear factor κB (NF-κB) activation, PDTC, diminishes U46619-induced VCAM-1 mRNA accumulation. NAC, which inhibits NF-κB and activation protein 1 (AP-1) binding activity, inhibits the expression of ICAM-1 or ELAM-1 at protein and mRNA levels. These findings suggest that ICAM-1 or ELAM-1 expression of HUVEC stimulated via TXA₂ receptors is augmented by induction of NF-κB and AP-1 binding activity through the PKC system, and that VCAM-1 expression is augmented by induction of NF-κB binding activity.     

核因子-κB对大鼠烧伤早期中性粒细胞在肺组织聚集中的作用

目的： 探讨大鼠烧伤早期肺组织核因子-κB（ NF-κB）活化对中性粒细胞(PMN)在肺组织中聚集和发生损害作用的影响。方法： 用Wistar大鼠 Ⅲ°35%TBSA烧伤模型。实验分正常大鼠对照组、烧伤组、烧伤后吡咯烷二硫代氨基甲酸盐（ PDTC）干预组。凝胶电泳迁移率分析法检测肺组织NF-κB活性；逆转录-聚合酶链式反应（RT-PCR）检测白细胞介素8（IL-8）和 细胞间黏附分子-1（ICAM-1） mRNA的表达；并测定肺组织髓过氧化物酶（MPO）活性和肺微血管von Willebrand因子（vWF)含量。结果： 大鼠烧伤后肺组织NF-κB活性在伤后1 h内即迅速增高，并持续增高到伤后24 h。伤后肺组织ICAM-1 和 IL-8 mRNA表达、MPO活性均明显高于、肺微血管vWF含量低于对照组。 PDTC处理显著缓解上述变化。结论： 严重烧伤后肺组织NF-κB活化，从而启动细胞黏附因子和趋化因子的合成和释放，导致PMN在肺组织中聚集，引起肺血管组织细胞损伤。     

脂多糖通过NF-κB途径上调大鼠腹膜间皮细胞表达CD40和ICAM-1

目的 观察脂多糖(LPS)作用下大鼠腹膜间皮细胞NF-κB活性及其对CD40和细胞间黏附分子1(ICAM-1)表达的影响.方法 分离及培养大鼠腹膜间皮细胞.LPS不同浓度作用12 h 及LPS (5 μg/ml)作用不同时间点收集细胞;LPS(5 μg/ml)或BAY11-7085(一种IκBα的磷酸化抑制剂)不同浓度(1 μmol/L和5 μmol/L)预处理3 h加LPS,作用3 h后收集细胞;采用RT-PCR方法检测CD40和ICAM-1 mRNA表达.采用蛋白印迹检测NF-κB和磷酸化NF-κB(p-NF-κB)蛋白表达.结果 与常规培养基对照组相比,5 μg/ml LPS组CD40和ICAM-1 mRNA表达显著升高(P<0.05),10 μg/ml LPS组显著高于5 μg/ml LPS作用组(P<0.05).5 μg/ml LPS 作用下,ICAM-1 mRNA表达从1 h开始升高,3 h达到高峰,之后逐渐降低.CD40 mRNA表达在1 h无显著变化,3 h时迅速达到高峰,之后逐渐降低.常规培养的大鼠腹膜间皮细胞结构性表达p-NF-κB蛋白;加入LPS后,p-NF-κB蛋白表达显著增加,其中30 min～1 h表达最强,之后逐渐降低,至2 h仍显著高于常规培养组(P<0.05).加入5 μmol/L BAY11-7085后,LPS诱导的CD40和ICAM-1 mRNA表达显著降低(P<0.05),与正常对照组相比差异无统计学意义. 结论 LPS以时间依赖和浓度依赖模式上调CD40和ICAM-1的表达.NF-κB信号途径参与调节LPS诱导的大鼠腹膜间皮细胞CD40和ICAM-1的表达.     

Effects of Citral on Lipopolysaccharide-Induced Inflammation in Human Umbilical Vein Endothelial Cells

Citral is an active compound of lemongrass oil which has been reported to have anti-inflammatory effects. In this study, we investigated the effects of citral on lipopolysaccharide (LPS)-induced inflammatory response in a rat model of peritonitis and human umbilical vein endothelial cells (HUVECs). LPS was intraperitoneally injected into rats to establish a peritonitis model. The HUVECs were treated with citral for 12 h before exposure to LPS. The levels of TNF-α and IL-8 were measured using ELISA. Western blotting was used to detect the expression of VCAM-1, ICAM-1, NF-κB, and PPAR-γ. The results showed that citral had a protective effect against LPS-induced peritonitis. Citral decreased the levels of WBCs and inflammatory cytokines TNF-α and IL-6. Citral also inhibited LPS-induced myeloperoxidase (MPO) activity in the peritoneal tissue. Treatment of HUVECs with citral significantly inhibited TNF-α and IL-8 expression induced by LPS. LPS-induced VCAM-1 and ICAM-1 expression were also suppressed by citral. Meanwhile, we found that citral inhibited LPS-induced NF-κB activation in HUVECs. Furthermore, we found that citral activated PPAR-γ and the anti-inflammatory effects of citral can be reversed by PPAR-γ antagonist GW9662. In conclusion, citral inhibits LPS-induced inflammatory response via activating PPAR-γ which attenuates NF-κB activation and inflammatory mediator production.     

葡萄糖胺对TNF-α诱导的血管内皮细胞VCAM-1表达的影响

目的探讨葡萄糖胺对肿瘤坏死因子-α(tumornecrosis factor-α,TNF-α)诱导的人脐带血管内皮细胞(HUVEC)血管细胞黏附分子(vascular cell adhesion molecule,VCAM-1)表达的影响。方法实时逆转录聚合酶链式反应(Real time RT-PCR)和免疫印迹法(Western blot)分别测定葡萄糖胺对VCAM-1的表达情况;Western blot法测定HUVEC核因子-κB(nuclear factor,NF-κB)磷酸化程度。结果葡萄糖胺抑制TNF-α诱导的VCAM-1在HUVEC的表达;NF-κB抑制剂BMS-345541抑制TNF-α诱导的VCAM-1的表达;葡萄糖胺抑制TNF-α诱导的NF-κB的磷酸化。结论葡萄糖胺抑制TNF-α诱导的VCAM-1的表达与抑制NF-κB的活化相关。