人颈动脉粥样硬化斑块环氧化物酶-2及I型前列腺素合成酶的表达

目的探讨环氧化物酶-2（cyclooxygenase type2，COX-2）及I型前列腺素合成酶（membrane associated prostaglandin E-1，mPGES-1）在人颈动脉粥样硬化斑块中的表达变化及作用机制。方法收集24例人颈动脉粥样硬化斑块标本和10例肠系膜动脉标本做对照组，应用免疫组织化学及逆转录PCR方法测定COX-2及mPGES-1mRNA表达水平，Western印记方法检测COX-2及mPGES-1的蛋白表达水平。比较不同程度动脉粥样硬化组织间COX-2、mPGES-1 mRNA表达水平及蛋白表达水平。结果颈动脉粥样硬化斑块组的免疫组织化学染色检测COX-2和mPGES-1呈阳性表达，斑块组COX-2 mRNA和mPGES-1 mRNA表达与对照组相比上调，差异有统计学意义（P〈0．05）；COX-2及mPGES-1 mRNA上调水平相关（P〈0．05）：颈动脉粥样硬化斑块的COX-2蛋白表达上调水平与对照组相比差异有统计学意义（P〈0．05）；颈动脉粥样硬化斑块COX-2、mPGES-1 mRNA及蛋白表达水平与病理损害程度有关，差异有统计学意义（P〈0．05）。结论COX-2及mPGES-1基因表达水平上调可能是进展性动脉粥样硬化损害的关键因素。     

急性脑梗死患者血清NOX2和MyD88蛋白水平与颈动脉粥样硬化斑块的相关性分析

目的 分析急性脑梗死患者血清中烟酰胺腺嘌呤二核苷酸磷酸氧化酶2(NOX2)和髓样分化因子88(MyD88)蛋白水平与颈动脉粥样硬化斑块的相关性。方法 选取2020年1月至2023年1月在宿迁市第一人民医院急诊抢救室需要急诊溶栓的80例急性脑梗死患者为观察组,及同期80例健康体检志愿者为对照组。经颈动脉彩超检查将患者分为无斑块组(18例)、稳定性斑块组(25例)、不稳定性斑块组(37例)。采用酶联免疫吸附(ELISA)法测定血清NOX2和MyD88蛋白水平。对患者血清NOX2和MyD88水平与颈动脉粥样硬化斑块的关系进行Spearman分析;受试者工作特征(ROC)曲线分析血清NOX2和MyD88蛋白水平对急性脑梗死患者不稳定性颈动脉粥样硬化斑块的诊断价值。结果 与对照组相比,观察组急性脑梗死患者血清中NOX2和MyD88水平均显著升高(P<0.05);无斑块组与稳定性斑块组患者血清NOX2和MyD88水平比较差异无统计学意义(P>0.05);不稳定性斑块组较无斑块组和稳定性斑块组患者血清NOX2和MyD88水平显著升高(P<0.05);急性脑梗死患者血清NOX2和My...     

急性脑梗死患者颈动脉粥样硬化斑块形成与血清脂蛋白a、MMP-9、D-D水平相关研究

目的 分析急性脑梗死患者颈动脉粥样硬化斑块形成与血清脂蛋白a、MMP-9、D-D水平的相关性.方法 选择2019年1月至2020年12月间收治的150例急性脑梗死患者作为观察组,100例脑梗死但未见颈动脉粥样硬化斑块患者作为对照组;入组后检测患者血糖、血脂、脂蛋白a、MMP-9及D-D水平.结果 观察组血糖、TG、TC、HDL及LDL水平均明显高于对照组,且差异具有统计学意义(P<0.05);观察组脂蛋白a、MMP-9及D-D水平均明显高于对照组,且差异具有统计学意义(P<0.05);脂蛋白a、MMP-9及D-D三指标均呈现明显的两两线性相关关系,且具有统计学意义(P<0.05);脂蛋白a、MMP-9及D-D水平与颈动脉粥样硬化斑块形成呈显著正相关关系,且具有统计学意义(P<0.01);脂蛋白a、MMP-9及D-D水平是影响颈动脉粥样硬化斑块形成的独立性危险因素(P<0.05).结论 急性脑梗死患者颈动脉粥样硬化斑块形成与血清脂蛋白a、MMP-9、D-D水平呈显著性正相关关系.     

瑞舒伐他汀通过抑制PI3K/AKT信号通路活性缓解小鼠颈动脉粥样硬化

目的探讨瑞舒伐他汀缓解小鼠颈动脉粥样硬化的机制及相关信号通路。方法将75只Apo E-/-小鼠随机分为5组,采取不同的干预措施,分别为对照组、颈动脉粥样硬化模型组、瑞舒伐他汀低剂量组、中剂量组、高剂量组。检测5组小鼠的血脂相关指标;实验结束后,处死小鼠,采用PCR技术,检测各组小鼠颈动脉组织中MMP-2 mRNA、MMP-9 mRNA的表达差异;采用Western-blot检测颈动脉组织中p-PI3K蛋白、p-AKT蛋白、PI3K蛋白、AKT蛋白的表达水平。结果①与模型组相比,3个瑞舒伐他汀治疗组小鼠的TC、TG、LDL-C水平明显下降,HDL-C明显升高(P0.05)。3个治疗组小鼠颈动脉总面积与模型组相比差异无统计学意义(P0.05),但斑块面积显著较小(P0.05),且高剂量组的斑块面积最小。②对照组小鼠颈动脉组织中MMP-2的阳性率较低,模型组的MMP-2的阳性率及染色强度明显较高。低、中、高剂量组的的染色阳性率及染色强度较模型组有明显下降。③模型组小鼠颈动脉组织中MMP-2 mRNA、MMP-9 mRNA的相对表达量显著最高,低剂量组次之,高剂量组表达水平最低。Western-blot检测结果趋势与RT-PCR一致。④与对照组相比,模型组小鼠颈动脉组织中p-PI3K蛋白、p-AKT蛋白表达量明显升高(P0.05)。与模型组比较,瑞舒伐他汀3个治疗组的小鼠颈动脉组织中p-PI3K蛋白、p-AKT蛋白表达量逐渐下降。结论瑞舒伐他汀可通过抑制PI3K/AKT信号通路活性对颈动脉粥样硬化小鼠发挥保护作用。     

通心络对颈动脉粥样硬化性斑块ABCAl表达的影响

目的:探讨通心络对兔模型颈动脉粥样硬化性斑块的三磷酸腺苷结合盒运转体Al (ABCAl)、视黄酸X受体(RXRα)信使核糖核酸(mRNA)表达的影响及机制。方法:32只新西兰大白兔分为4组,其中空白对照组8只(A组),余24只兔于右侧颈动脉放置改良的硅橡胶圈,加1%高胆固醇喂养的方法建立粥样硬化性颈动脉狭窄动物模型。颈动脉狭窄模型而无药物干预的对照组8只(B组);辛伐他丁治疗组8只,给予辛伐他丁5 mg/kg,每天1次(C组);通心络治疗组8只,给予通心络1 g/kg,每日1次(D组)。药物干预前后分别检测兔模型静脉血的血清甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL)及高密度脂蛋白胆固醇(HDL)水平,两种药物干预2周后处死动物取右侧颈动脉狭窄段及对侧相应段血管,以Western Blot法测定其ABCAl、RXRα蛋白质表达量。结果:给予通心络和辛伐他丁干预兔模型组(C组和D组)ABCAl、RXRα蛋白质表达水平均增高,血脂水平下降。结论:通心络可使模型兔颈动脉粥样硬化斑块的ABCAl mRNA、RXRαmRNA表达水平和其蛋白质表达水平上调,可能有益于抗动脉粥样硬化斑块形成作用。     

非酒精性脂肪肝患者血清FGF21水平变化与颈动脉粥样硬化斑块发生的相关性

目的:探讨非酒精性脂肪肝(NAFLD)患者血清成纤维细胞生长因子21(FGF21)水平变化及与颈动脉粥样硬化斑块发生的相关性。方法:选取2023-01-2023-09确诊的NAFLD患者131例,根据是否存在颈动脉粥样硬化斑块分为无斑块组(n=59)和斑块组(n=72),选取同期体检健康者39例为对照组。检测所有对象血清尿酸(UA)、低密度脂蛋白胆固醇(LDL-C)、小而密低密度脂蛋白胆固醇(sdLDL-C)、高密度脂蛋白胆固醇(HDL-C)、甘油三酯(TG)、总胆固醇(TC)和FGF21水平并比较组间差异。Logistic回归分析FGF21与NAFLD患者颈动脉粥样硬化斑块发生的相关性。结果:与对照组比较,无斑块组和斑块组患者UA、TC、TG、LDL-C、sdLDL-C和FGF21水平均升高,HDL-C水平降低(P<0.05或P<0.01);与无斑块组比较,斑块组LDL-C、sdLDL-C和FGF21升高(P<0.05或P<0.01)。Logistic回归分析显示,在充分调整协变量后,血清FGF21升高与NAFLD患者动脉粥样硬化斑块发生呈正相关。结论:血清F...     

2型糖尿病合并颈动脉粥样硬化患者外周血单个核细胞凋亡相关斑点样蛋白mRNA的表达

目的:分析2型糖尿病(T2DM)合并颈动脉粥样硬化(CAS)患者外周血单个核细胞(PBMCs)中衔接蛋白凋亡相关斑点样蛋白(ASC)mRNA的表达及作用。方法:根据华盛顿大学CAS斑块超声分级标准将107例T2DM患者分为单纯T2DM组(n=26)、T2DM轻度斑块组(n=32)、T2DM中度斑块组(n=38)和T2DM重度斑块组(n=11)。另选非T2DM的CAS患者作为单纯CAS组(n=35),以及体检健康者作为正常对照组(n=35)。采用实时荧光定量聚合酶链反应(RT-FQ-PCR)技术检测各组PBMCs中ASC mRNA表达水平。统计分析各组差异及ASC mRNA水平与CAS斑块严重程度的关系。结果:各病例组ASC mRNA表达水平显著高于正常对照组(P0.05),T2DM合并CAS组ASC mRNA表达水平显著高于单纯CAS组(P0.01);ASC mRNA表达水平轻度斑块组中度斑块组重度斑块组,即ASC mRNA表达水平与T2DM患者CAS斑块程度呈明显负相关(r=-0.43,P0.05)。结论:ASC mRNA表达增加可能是T2DM合并AS的发病因素和CAS斑块活跃性的指征。     

人冠状动脉内半乳糖凝集素3水平与斑块稳定性的相关性研究

目的:探讨人冠状动脉粥样斑块内半乳糖凝集素3(galectin-3)的表达水平与斑块结构及稳定性的关系。方法:收集尸检案例的冠状动脉标本84例,其中动脉粥样硬化但非冠心病猝死者22例(A1组),冠心病猝死但不伴有继发病变者20例(A2组),冠心病猝死且伴有继发病变者24例(A3组),无心脏疾病死亡者18例作为正常对照组(control组)。所有冠状动脉行常规HE染色检测内膜厚度、坏死核心厚度、纤维帽厚度和血管狭窄程度;以单核巨噬细胞表面标志物CD68标记病灶内的单核-巨噬细胞并计数,采用免疫组织化学、Western blot及RT-qPCR法检测冠状动脉组织中galectin-3、CD68和基质金属蛋白酶2(MMP-2)表达,并分析其表达与斑块结构及其稳定性的关系。结果:同正常组比较,病变各组粥样硬化病灶内的内膜和坏死核心增厚,纤维帽变薄,血管狭窄程度增高(P0.05);病灶内泡沫细胞数量增多(P0.05);病灶内galectin-3、CD68和MMP-2蛋白及mRNA水平较正常组显著升高,且在A1组、A2组和A3组中呈递增趋势(P0.05);病灶内galectin-3、CD68和MMP-2的表达与内膜厚度和坏死灶厚度呈正相关,与纤维帽厚度呈负相关。结论:人冠状动脉粥样硬化病灶内galectin-3表达升高,并与斑块结构稳定性相关。     

脂蛋白磷脂酶A2在颈动脉粥样硬化患者中的临床应用价值

目的 探讨血浆脂蛋白磷脂酶A2(lipoprotein-associated phospholipase A2,Lp-PLA2)在颈动脉粥样硬化患者中的临床应用价值.方法 用ELISA法检测132例颈动脉粥样硬化患者(Crouse斑块积分Ⅰ级34例,Ⅱ级31例,Ⅲ级32例,Ⅳ级35例;稳定斑块组85例,不稳定斑块组47例)、129例正常健康对照者血浆Lp-PLA2水平,用高分辨超声测定颈动脉粥样硬化患者颈动脉内膜中层厚度(intima-media thickness,IMT),分析颈动脉粥样硬化患者Lp-PLA2水平与IMT、Crouse斑块积分间的相关性.结果 颈动脉粥样硬化组的血浆Lp-PLA2水平(249.44±19.62 ng/mL)、IMT(1.68 ±0.19mm)明显高于正常对照组(151.63±11.89 ng/mL,P＜0.01;0.53 ±0.04 mm,P＜0.01);Crouse斑块积分I级Lp-PLA2水平(189.51±14.20 ng/mL)、IMT(1.45 ±0.14mm)与Ⅱ级Lp-PLA2水平(206.27±14.93 ng/mL,P＞0.05)、IMT(1.51±0.16mm,P＞0.05)的水平差异无统计学意义,Crouse斑块积分Ⅱ级与Ⅲ级、Ⅱ级与Ⅳ级以及Ⅲ级与Ⅳ级Lp-PLA2、IMT的水平差异有统计学意义(P ＜0.01或P＜0.05).颈动脉粥样硬化患者中不稳定斑块组血浆Lp-PLA2水平(263.12±17.60ng/mL)、IMT(1.87 ±0.20mm)高于稳定斑块组(225.31±15.47 ng/mL,P＜0.05;1.59±0.17mm,P＜0.05);单因素相关性分析显示,IMT与Lp-PLA2呈正相关(r=0.391,P＜0.05).结论 Lp-PLA2水平与IMT、Crouse斑块积分呈正相关,具有较好的临床应用价值.     

兔股动脉粥样硬化斑块模型中TF及TFPI的变化

目的： 观察兔股动脉粥样硬化斑块发展过程中斑块内组织因子（TF）及组织因子途经抑制物-1（TFPI-1）、组织因子途经抑制物-2（TFPI-2）的变化。方法： 新西兰雄兔通过高脂饲养合并股动脉球囊损伤建立动脉粥样硬化斑块模型。在建模8周、10周和12周，分别截取球囊损伤处股动脉标本，检测斑块组织内TF、TFPI-1和TFPI-2的蛋白表达；并测定斑块内TF、TFPI-1和TFPI-2的mRNA水平。结果： 自8周、10周到12周，血管斑块中TF、TFPI-1和TFPI-2的观测区域内阳性染色面积均逐渐增加。自8周、10周到12周，血管斑块中TF的mRNA水平和TFPI-1的mRNA水平均逐渐增加；TFPI-2mRNA表现为前期无明显变化，12周时显著下调。结论： 在兔股动脉粥样硬化斑块的形成过程中，随着斑块内TF基因及蛋白表达的增加,TFPI-1和TFPI-2反应性表达增加，但其表达水平未能完全抑制斑块的进展。     

α-硫辛酸对兔动脉粥样硬化斑块CD147及基质金属蛋白酶的表达的影响

目的 探讨α-硫辛酸对兔动脉粥样硬化斑块形成及稳定性影响的机制.方法 采用液氮冻伤术联合高脂饲料饲养构建兔动脉粥样硬化模型,并使用α-硫辛酸处理动物2周,采用HE染色检测颈动脉组织的病理状态;ELISA检测给药前后兔血清中MMP-2和MMP-9的含量;Real-Time PCR和Western印迹检测给药后动脉组织中MMP-2和MMP-9的表达,并用Real-Time PCR检测组织中CD147的mRNA表达.结果 模型兔颈总动脉组织出现斑块,脂质沉积,泡沫细胞增多,炎性细胞浸润,α-硫辛酸能明显改善这种病理情况.模型兔血清中MMP-2和MMP-9的含量明显高于对照组(P<0.05),α-硫辛酸处理2周后模型兔血清中MMP-2和MMP-9较模型组明显下降(P<0.05).各组动物动脉组织中MMP-2和MMP-9的mRNA和蛋白表达水平与血清中变化趋势一致,均具有显著差异(P<0.05).α-硫辛酸处理后动脉粥样硬化兔的动脉组织中CD147的mRNA水平也较模型组明显下调(P<0.05).结论 α-硫辛酸可能是通过下调CD147的方式来降低基质金属蛋白酶MMP-2和MMP-9的表达,最终改善兔动脉粥样硬化的进程.     

脑梗死患者间歇使用低分子肝素对颈动脉斑块和血清炎性标志物的影响

目的:脑梗死患者间断使用低分子肝素,观察其对颈动脉斑块和血清炎性标志物的影响.方法:选择脑梗死并具有颈动脉斑块的患者78例作为对象,随机分为常规组(37例)和低分子肝素组(41例),分别于治疗前后测定颈动脉斑块情况及血清高敏C反应蛋白(hs-CRP)、基质金属蛋白酶-9(MMP-9)水平.结果:①治疗前比较,两组间颈动...     

慢病毒介导的SHP-1过表达抑制模型小鼠动脉粥样硬化

目的:研究含SH2结构域的酪氨酸磷酸酶-1(SHP-1)慢病毒载体在动脉粥样硬化模型小鼠中的作用。方法:将45只8周龄雄性Apo E基因敲除小鼠随机分为3组,即对照组、绿色荧光蛋白(GFP)转染组、SHP-1转染组。3组小鼠右侧颈总动脉植入套环并高脂喂养8周,随后分别转染GFP空载体和SHP-1慢病毒(SHP-1-LV)。分别于转染第1、2、6周检测硬化斑块荧光强度、小鼠体重、血清总胆固醇(TC)、甘油三酯(TG)水平变化,并利用real-time PCR和Western blot检测右侧颈总动脉中SHP-1、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、基质金属蛋白酶(MMP)-2、MMP-9等的表达,最后制作切片染色观察斑块病理变化。结果:荧光显微镜下观察到慢病毒转染第1、2、6周后斑块有明显荧光,且在第2周时荧光强度最高,SHP-1慢病毒转染对小鼠体重和血清TC、TG水平无显著影响,但可显著上调右侧颈总动脉中SHP-1的mRNA和蛋白的表达,同时抑制IL-6、TNF-α、MMP-2、MMP-9等的表达水平。SHP-1慢病毒转染也降低右侧颈总动脉斑块面积比及脂质含量。结论:过表达SHP-1可能促进小鼠动脉粥样硬化斑块消退,从而为预防和治疗动脉粥样硬化提供靶点。     

动脉粥样硬化中CARHSP1基因的表达对缺氧诱导的血管内皮细胞活力凋亡及免疫因子的影响

目的:探讨动脉粥样硬化斑块中钙调节热稳定蛋白1(CARHSP1)基因的表达对缺氧诱导的血管内皮细胞活力、凋亡及白细胞介素6(IL-6)和C-反应蛋白(CRP)表达的影响。方法:用Western blot检测动脉粥样硬化斑块中CARHSP1的蛋白表达;缺氧处理人脐静脉内皮细胞(HUVECs),将细胞分为正常培养组、缺氧组、缺氧+CARHSP1-siRNA组和缺氧+pc DNA3. 1-CARHSP1组,用CCK-8法及流式细胞术分别检测各组细胞活力及凋亡率; RT-PCR检测IL-6和CRP的表达; Western blot检测凋亡蛋白caspase-3、cleaved caspase-3、Bcl-2和Bax的蛋白水平。结果:CARHSP1在动脉粥样硬化斑块中的蛋白表达显著高于对照组(P 0. 05);缺氧可明显增加CARHSP1的表达。缺氧组HUVECs活力及Bcl-2表达显著低于正常培养组,细胞凋亡率及IL-6、CRP、cleaved caspase-3和Bax的蛋白水平显著高于正常培养组(P 0. 05);与缺氧组比较,缺氧+CARHSP1-siRNA组的活力及Bcl-2表达显著升高,细胞凋亡率及IL-6、CRP、cleaved caspase-3和Bax的蛋白水平显著降低(P 0. 05),pc DNA3. 1-CARHSP1组的细胞活力及Bcl-2表达显著降低,细胞凋亡率及IL-6、CRP、cleaved caspase-3和Bax表达显著升高(P 0. 05)。结论:CARHSP1在动脉粥样硬化斑块中表达升高,抑制CARHSP1表达可提高HUVECs的活力,降低细胞凋亡,下调免疫因子IL-6和CRP的表达,而过表达CARHSP1则反之。     

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。     

Expression of cyclooxygenase-1/-2, microsomal prostaglandin-E synthase-1 and E-prostanoid receptor 2 and regulation of inflammatory mediators by PGE2 in the amoeboid microglia in hypoxic postnatal rats and murine BV-2 cells

This study aimed to investigate the effect of hypoxia on the expression of cyclooxygenase-1 (COX-1), cyclooxygenase-2 (COX-2), microsomal prostaglandin-E synthase (mPGES-1), E-prostanoid receptor 2 (EP2) in microglia; and the roles of EP2-cyclic adenosine monophosphate (cAMP) signaling pathway in the prostaglandin E₂ (PGE₂) regulation of inflammatory mediators released by hypoxic BV-2 cells. Immunoexpression of COX-1, COX-2, mPGES-1 and EP2 was localized in the amoeboid microglial cells (AMC), a nascent brain macrophage in the developing brain, as confirmed by double labeling with OX-42 and lectin, specific markers of microglia. AMC emitted a more intense immunofluorescence in hypoxic rats when compared with the matching controls. In postnatal rats subjected to hypoxia, mRNA and protein expression levels of COX-1, COX-2 and mPGES-1 along with tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), inducible nitric-oxide synthase (iNOS) and PGE₂ product in the callosal tissue were significantly increased. The results were shared in the BV-2 cells except for COX-1 mRNA and protein whose levels remained unaltered. Interestingly, treatment with EP2 antagonist AH-6809 resulted in suppression of hypoxia induced EP2, IL-1β and iNOS mRNA and protein expression, TNF-α protein expression and intracellular cAMP level in BV-2 cells. It is suggested that PGE₂ may regulate above inflammatory mediators in the activated microglia via EP2-cAMP signaling pathway in hypoxic conditions.     

Microsomal prostaglandin E synthase (mPGES)-1, mPGES-2 and cytosolic PGES expression in human gastritis and gastric ulcer tissue

Recently, three different prostaglandin E2 synthases have been identified: microsomal prostaglandin E synthase (mPGES)-1, cytosolic PGES (cPGES), and mPGES-2; however, their role and connection to cyclooxygenase (COX)-2 in the gastric ulcer repair process remain unknown. Therefore, we examined mPGES-1, cPGES, and mPGES-2 expression and localization in the stomach in vitro and in vivo. Tissues were obtained from Helicobacter pylori (H. pylori)-infected patients and consisted of surgical resections of gastric ulcers, or biopsies of gastric ulcers or gastritis. mPGES-1 mRNA and protein expression levels were examined by real-time polymerase chain reaction (PCR) and Western blot analysis, respectively. mPGES-1, cPGES, and mPGES-2 localization were analyzed immunohistochemically. Induction of PGES expression in response to interleukin (IL)-1beta was examined in vitro in the cultured human gastric fibroblast line Hs262.St. Real-time PCR analysis of mPGES-1 mRNA expression in biopsy samples showed significantly higher expression levels in open than in closed gastric ulcer tissue. Western blot analysis showed mPGES-1 protein expression limited to open ulcer tissue, while mPGES-2 and cPGES immunoreactivities were seen in both open and closed ulcer tissue. Immunohistochemical analysis showed strong mPGES-1 expression in fibroblasts and macrophages of the ulcer bed, paralleling COX-2 expression. cPGES and mPGES-2 expression levels were seen in both fibroblasts of the ulcer bed and in epithelial cells. Furthermore, stronger cPGES and mPGES-2 immunoreactivities were seen in scattered mast cell-like cells and neuroendocrine-like cells, respectively. Induction of mPGES-1 expression in response to IL-1beta was seen in cultured gastric fibroblasts in vitro, and double immunostaining showed mPGES-1 coexpression with COX-2 in fibroblasts of the ulcer bed in vivo. In conclusion, mPGES-1, cPGES, and mPGES-2 are all expressed in gastric ulcer tissue, but only mPGES-1 parallels COX-2 expression in mesenchymal and inflammatory cells of the ulcer bed, suggesting a key role for this enzyme in the ulcer repair process.     

Induction of Microsomal Prostaglandin E Synthase-1 in Human Gingival Fibroblasts

It is well established that prostaglandin E2 (PGE2) plays an important role in inflammatory diseases including periodontitis. Previously we have reported that the inflammatory mediators interleukin-1beta, (IL-1beta) and tumor necrosis factor alpha (TNFalpha) stimulate PGE2 synthesis by inducing mRNA expression of cyclooxygenase-2 (COX-2) in human gingival fibroblasts. In present study the involvement of microsomal prostaglandin E synthase-1 (mPGES-1) in relation to PGE2 production was investigated. The results showed that IL-1beta as well as TNFalpha induced mPGES-1 mRNA and protein expression accompanied by enhanced PGE2 production in gingival fibroblasts. The anti-inflammatory steroid dexamethasone (DEX) inhibited mPGES-1 mRNA and protein expression as well as PGE2 production induced by IL-1beta or TNFalpha. The COX-2 specific inhibitor, celecoxib, in contrast to the nonspecific COX inhibitor, indomethacin, markedly reduced mPGES-1 expression induced by IL-1beta. The results demonstrate that mPGES-1 regulates PGE2 production in gingival fibroblasts stimulated by inflammatory mediators IL-1beta and TNFa. This novel pathway may be a potential target for treatment strategies of periodontal disease.     

Expression and regulation of prostaglandin E synthase isoforms in human myometrium with labour

Since the controversies regarding the use of non-steroidal anti-inflammatory drugs (NSAIDs) and selective cyclo-oxygenase (COX)-2 antagonists for the treatment of preterm labour (PTL), more emphasis has been placed on investigating the terminal synthases involved in the production of prostaglandins (PGs) to allow more targeted therapy in PTL. Prostaglandin E(2) (PGE(2)) is synthesized by one of three enzymes, cytosolic prostaglandin E synthase (cPGES), microsomal PGES-1 (mPGES-1) and microsomal PGES-2 (mPGES-2). We have determined (i) the immuno-localization of all three PGES enzymes in lower segment pregnant human myometrium, (ii) the expression of PGES and COX-2 mRNA expression at term and preterm gestation with and without labour and (iii) the effect of interleukin (IL)-1beta on COX-2 and PGES mRNA and protein expression in human myometrial smooth muscle (HMSM) cell cultures. We show mPGES-1 protein located predominantly in myometrial and vascular smooth muscle cells (SMCs), whilst mPGES-2 protein is largely in stromal cells surrounding the SMC and cPGES is diffusely located throughout the myometrium. Expression of mPGES-2 mRNA increased with term labour and PTL and expression of COX-2 and mPGES-1 mRNA with term labour, whereas cPGES expression did not change. IL-1beta stimulated release of PGE(2) by HMSM cells and increased COX-2 and mPGES-1 mRNA and protein expression. Thus, COX-2 expression and mPGES-1 expression are co-ordinately up-regulated in lower segment myometrium with term labour and with IL-1beta treatment in HMSM cells.