LPS对MRL/MpJ小鼠脾T细胞表达TLR4和TNFα的影响

研究Toll样受体 4 (TLR4 )在MRL/MpJ小鼠脾细胞的表达状况和革兰阴性菌脂多糖 (LPS )刺激后TLR4的表达变化以及对TNF α产生的影响。MRL/MpJ小鼠经腹腔注射LPS后在不同时间取脾细胞 ,用三色荧光标记流式细胞仪技术检测小鼠脾T细胞TLR4的表达情况 ,并与BALB/c小鼠作对照 ;通过RT PCR观察LPS刺激前后脾T细胞TNF αmRNA的表达变化。结果表明TLR4在MRL/MpJ小鼠CD3+ CD4 + T细胞和CD3+ CD8+ T细胞中均有表达 ;但TLR4 + /CD4 + 细胞表达仅BALB/c小鼠的 33% ,在受LPS刺激后 ,MRL/MpJ鼠CD4 + T细胞TLR4升高时间较BALB/c小鼠延迟 ;CD8+ T细胞TLR4在LPS刺激前后无明显变化 ;脾细胞TNF αmRNA水平升高亦延迟。通过对干燥综合征样自身免疫病鼠模型MRL/MpJ小鼠脾T细胞TLR4的表达及功能研究 ,将有助于认识天然免疫和获得性免疫间的联系和感染在自身免疫病发病中的作用。     

CD39/NTPDase-1 activity and expression in normal leukocytes

INTRODUCTION: CD39/NTPDase-1 is a cell surface enzyme expressed on leukocytes and endothelial cells that metabolizes ATP to ADP and AMP. CD39 is expressed on numerous different types of normal leukocytes, but details of its expression have not been determined previously. METHODS: We examined CD39 expression and activity in leukocytes isolated from healthy volunteers. Expression of CD39 on leukocytes was measured by FACS and activity of CD39 in lymphocytes and neutrophils was determined by an enzymatic radio-TLC assay. RESULTS: We established that CD39 is expressed on neutrophils, lymphocytes, and monocytes. The enzyme is found on >90% of monocytes, neutrophils, and B-lymphocytes, and 6% of T-lymphocytes and natural killer cells. Per cell density of expression varied, with the highest expression on monocytes and B-lymphocytes. ATPase and ADPase activities were highest on B-lymphocytes, lower on neutrophils, lowest on T-lymphocytes. The ratio of ADPase:ATPase activity was 1.8 for neutrophils and B-lymphocytes and 1.4 for T-lymphocytes. Hypertensive volunteers had lower levels of CD39 on their T-lymphocytes and NK cells. No correlation between age, gender, ethnic background, or cholesterol level and CD39 expression was observed. CONCLUSIONS: We conclude that CD39 activity and expression are present to varying degrees on all leukocytes types examined. Differences between leukocyte types should be considered when examining CD39 in disease states.     

注射用炎琥宁对脂多糖发热模型家兔解热机制的研究

目的 研究注射用炎琥宁对脂多糖发热模型家兔血清中细胞因子（TNF-α、IL-1β、IL-6、cAMP、PGE2）和下丘脑中升温介质（cAMP、PGE2）的影响。方法 建立家兔脂多糖（10 EU/kg）发热模型,监测不同时间点注射用炎琥宁对脂多糖发热模型家兔体温影响,绘制平均升温曲线;模型升温峰值时收集指标,ELISA法检测血清中细胞因子、下丘脑中升温介质变化。结果 注射用炎琥宁能显著降低脂多糖发热模型家兔的体温,且量效关系较显著;注射用炎琥宁能显著抑制发热模型家兔血清中IL-6、cAMP、PGE2增加和下丘脑中cAMP、PGE2的释放。结论 注射用炎琥宁对脂多糖发热模型家兔有较好的解热作用,其解热效果可能与其抑制外周IL-6、cAMP、PGE2增加和下丘脑cAMP、PGE2释放有关。     

TNF-α、IL-8在脂多糖致大鼠脑水肿中的表达

目的　探讨脂多糖 (lipopolysaccharide ,LPS)致大鼠脑水肿发生过程中 ,地塞米松作用下TNF α、IL 8表达水平的变化及相互关系。方法　利用LPS诱导大鼠脑水肿发生 ,测定脑组织含水量及伊文私蓝 (Evansblue,EB)含量 ,制备脑组织匀浆 ,ELISA法测IL 8含量 ,放射免疫法测TNF α含量。结果　LPS注入后LPS组和DXM组脑组织含水量和伊文思蓝含量均较NS组增高 (P <0 .0 5) ;LPS组TNF α和IL 8含量较NS组显著增高 (P <0 .0 1 ) ,但DXM组较LPS组显著降低 (P <0 .0 5) ;DXM组TNF α和IL 8含量在早期 (≤ 2 4h)较NS组增高 (P <0 .0 5) ,48h时则无显著差异 (P >0 .0 5)。LPS组脑组织含水量与EB含量 ,TNF α与脑组织含水量 ,IL 8与脑组织含水量 ,IL 8与EB含量均呈正相关 (r值分别为 0 .537,0 .42 3 ,0 .473 ,0 .831 ,P <0 .0 5)。结论　TNF α、IL 8参与了脂多糖诱导的脑水肿的发生、发展过程 ,早期应用糖皮质激素可以抑制炎性细胞因子的生成 ,减轻炎症反应与组织损伤。     

地塞米松对脂多糖诱导急性肺损伤大鼠的保护作用

目的 通过地塞米松对脂多糖（Lipolysaccharides,LPS）诱导的大鼠急性肺损伤时炎症介质的影响作用,探讨其对肺脏的保护机制。方法 健康雄性S—D大鼠随机分成A、B、C、D组,各组分别按1、2、4h三个时间点再设立3个亚组。A组经腹腔注射地塞米松;B组经腹腔注射LPS,C组在腹腔注射LPS前1h经腹腔注射地塞米松,D组为对照组。在到达各时间点后,经腹腔注射加倍剂量的硫喷妥钠处死,切开气管,并予0℃0．9％氯化钠注射液（normal saline,NS）作肺泡灌洗。采用酶联吸附免疫法（ELISA）测定肺泡灌洗液（broncho alveokar lavage fluid,BALF）中的肿瘤坏死因子-α（tumor necrosis factor—α,TNF—α）、白介素-1β（iaterlrukin—1β,IL—1β）与巨噬细胞炎性蛋白-2（macrophafe inflamnatovy protein-2,MIP-2）。另外12只大鼠按上述分组法随机分成4组,每组3只,动物模型同上,作肺组织病理观察。结果 在病理形态学上与D组比较,B组肺损伤严重。BALF中TNF—α、IL—1β、MIP-2在A、B、C、D组间比较差异均有统计学意义（F分别=15．51、47．37、19．94、134．27、65．44、57．29、18．44、85.31、89.13,P均〈0．05）。B组与D组比较,TNF—α差异有统计学意义（t分别=5．37、8.69、6．43,P均〈0．05）、IL—1β差异有统计学意义（t分别=7．25、9.77、11．51,P均〈0．05）、MIP-2差异有统计学意义（t分别=6.84、12.38、15．52,P均〈0．05）;C组与D组比较,TNF—α差异有统计学意义（t值分别为2．43、2．67、1．98,P均〈0．05）、IL—1β差异有统计学意义（t分别=3．58、4．15、4．81,P均〈0.05）、MIP-2差异有统计学意义（t分别=1．87、3．73、7．55,P均〈0．05）;C组与B组比较,TNF—α差异有统计学意（t分别：3．15、4．19、4．03,P均〈0．05）、IL—1β差异有统计学意（t分别=3．71、5．27、5．81,P均〈0．05）、MIP-2差?     

LPS对MRL／MpJ小鼠脾T细胞表达TLR4和TNF-a的影响

研究Toll样受体4(TLR4)在MRL／MpJ小鼠脾细胞的表达状况和革兰阴性菌脂多糖(LPS)刺激后TLR4的表达变化以及对TNF-a。产生的影响。MRL／MpJ小鼠经腹腔注射LPS后在不同时间取脾细胞．用三色荧光标记流式细胞仪技术检测小鼠脾T细胞TLR4的表达情况，并与BALB／c小鼠作对照；通过RT-PCR观察LPS刺激前后脾T细胞TNF-dmRNA的表达变化。结果表明TLR4在MRL／MpJ小鼠CD3^ CD4^ T细胞和CD3^ CD8^ T细胞中均有表达；但TLR4^ ／CD4^ 细胞表达仅BALB／c小鼠的33％，在受LPS刺激后，MRL／MpJ鼠CD4^ T细胞TLR4升高时间较BALB／c小鼠延迟；CD8^ T细胞TLR4在LPS刺激前后无明显变化；脾细胞TNF-amRNA水平升高亦延迟。通过对干燥综合征样自身免疫病鼠模型MRL／MpJ小鼠脾T细胞TLR4的表达及功能研究，将有助于认识天然免疫和获得性免疫间的联系和感染在自身免疫病发病中的作用。     

哌唑嗪抑制脂多糖诱导的磷脂酶A2活性 及肝细胞内钙离子超载

目的：研究α－肾上腺素受体拮抗剂哌唑嗪对脂多糖（LPS）诱导的大鼠血清中磷脂酶A2活性及肝细胞内钙离子浓度[Ca^2 ]i超载的抑制及其对内毒素性肝损害的保护作用及机制。方法：用腹腔注射LPS（2mg/kg)复制急性肝损害型，用哌唑嗪（4mg/kg)灌胃为保护组；检测了处置6小时后各组大鼠肝细胞内[Ca^2 ]i，24小时后血清丙氨酸转氨酸转氨酶、天冬氨酸转氨酶、乳酸脱氨酶、γ－谷氨酰转肽酶及肝脏病理切片；并对各组血清中分泌型磷脂酶A2（sPLA2)活性，前列腺素E2（PGE2）肿瘤坏死因子－α（TNF-α）的含量进行了动态测定。结果：哌唑嗪明显降低了由LPS诱导的肝细胞[Ca^2 ]i升高和肝功能损害，减轻了肝细胞坏死程度和炎性浸润，同时对LPS所引起的血清中sPLA2,PGE2和TNF－α的升高均有不同程度的抑制。结论：哌唑嗪可通过降低LPS诱导的肝细胞内Ca^2 超载、减少TNF－α的产生从而抑制sPLA的过度激活及炎症介质的产生，对肝脏具有明显的保护作用。     

Inhibition of stress-activated MAP kinases induces clinical improvement in moderate to severe Crohn's disease.

BACKGROUND & AIMS: We investigated if inhibition of mitogen-activated protein kinases (MAPKs) was beneficial in Crohn's disease. METHODS: Inhibition of JNK and p38 MAPK activation with CNI-1493, a guanylhydrazone, was tested in vitro. Twelve patients with severe Crohn's disease (mean baseline, CDAI 380) were randomly assigned to receive either 8 or 25 mg/m(2) CNI-1493 daily for 12 days. Clinical endpoints included safety, Crohn's Disease Activity Index (CDAI), Inflammatory Bowel Disease Questionnaire, and the Crohn's Disease Endoscopic Index of Severity. RESULTS: Colonic biopsies displayed enhanced JNK and p38 MAPK activation. CNI-1493 inhibition of both JNK and p38 phosphorylation was observed in vitro. Treatment resulted in diminished JNK phosphorylation and tumor necrosis factor production as well as significant clinical benefit and rapid endoscopic ulcer healing. No serious adverse events were noted. A CDAI decrease of 120 at week 4 (P = 0.005) and 146.5 at week 8 (P = 0.005) was observed. A clinical response was seen in 67% of patients at 4 weeks and 58% at 8 weeks. Clinical remission was observed in 25% of patients at week 4 and 42% at week 8. Endoscopic improvement occurred in all but 1 patient. Response was seen in 3 of 6 infliximab failures, 2 of whom showed remission. Fistulae healing occurred in 4 of 5 patients, and steroids were tapered in 89% of patients. CONCLUSIONS: Inflammatory MAPKs are critically involved in the pathogenesis of Crohn's disease and their inhibition provides a novel therapeutic strategy.     

人角膜基质成纤维细胞分泌作用与脂多糖的影响

背景：体外培养的角膜基质成纤维细胞,可以识别革兰阴性细菌细胞壁成分脂多糖,并释放多种细胞因子和趋化因子,从而促进角膜溃疡的形成。 目的：观察细菌脂多糖对角膜基质成纤维细胞分泌作用的影响。 方法：体外培养人角膜基质细胞,取第3~5代细胞鉴定后用于实验。采用ELISA法检测角膜基质细胞脂多糖作用前后培养上清中分泌白细胞介素1,肿瘤坏死因子α,白细胞介素6,白细胞介素8水平。 结果与结论：脂多糖在一定质量浓度范围及作用时间内,对人角膜基质细胞增殖存活率没有影响。培养的人角膜基质成纤维细胞不表达白细胞介素1,肿瘤坏死因子α,未给予脂多糖的角膜基质细胞微量表达白细胞介素6,白细胞介素8,脂多糖(100 μg/L)作用24 h后,白细胞介素6,白细胞介素8水平明显增高。