异丙酚对LPS诱导BV-2小胶质细胞IL-1β和TNF-α释放的影响及TLR4在其中的作用

目的 评价异丙酚对LPS诱导BV-2小胶质细胞IL-1β和TNF-α释放的影响及Toll样受体4(TLR4)在其中的作用.方法 将体外培养的BV-2小胶质细胞接种于96孔培养板中,采用随机数字表法,将其随机分为4(n=12):对照组、LPS组、异丙酚组和LPS+异丙酚组.LPS组加入LPS1μg/ml孵育24h;异丙酚组加入异丙酚30 μmol/L孵育24 h;LPS+异丙酚组同时加入LPS 1 μg/ml和异丙酚30 μmol/L孵育24h.于孵育6h时,采用ELISA法检测细胞上清液TNF-α浓度,以此反映TNF-α的释放量,采用RT-PCR法测定TLR4 mRNA表达;于孵育24h时,采用ELISA法检测细胞上清液IL-1β浓度,以此反映IL-1β的释放量,采用Western Blot法检测TLR4蛋白表达.结果 与C组比较,LPS组和LPS+异丙酚组IL-1β和TNF-α的释放量升高,TLR4 mRNA及其蛋白表达上调(P＜0.05);与LPS组比较,LPS+异丙酚组IL-1β和TNF-α的释放量降低,TLR4 mRNA及其蛋白表达下调(P＜0.05).结论 异丙酚可抑制LPS诱导BV-2小胶质细胞IL-1β和TNF-α的释放,其机制与抑制TLR4的表达有关.     

异丙酚不同时机给药对大鼠海马缺氧复氧损伤神经元胞浆细胞色素c的影响

目的 评价异丙酚不同时机给药对大鼠海马缺氧复氧损伤神经元胞浆细胞色素c浓度(Cty c)的影响.方法 原代培养大鼠海马神经元,采用随机数字表法,将其随机分为5组(n=5):对照组(C组)、缺氧复氧损伤模型组(M组)和异丙酚不同时机给药组(Ⅰ组、Ⅱ组和Ⅲ组).采用缺氧6h再复氧的方法制备海马神经元缺氧复氧损伤模型.Ⅰ组、Ⅱ组和Ⅲ组分别于缺氧前、复氧即刻和复氧2 h(T0～2)时加入异丙酚至终浓度20 μmol/L.各组分别于T1,2和复氧24 h(T3)时观察细胞凋亡情况,检测胞浆Cytc的浓度.结果 与C组相比,M组T1-3时、Ⅰ组、Ⅱ组和Ⅲ组T1,2时细胞胞浆Cty c浓度升高(P＜0.05);与M组相比,Ⅰ组T1-3时、Ⅱ组和Ⅲ组T1.2时细胞胞浆Cty c浓度降低(P＜0.05);与Ⅰ组相比,Ⅱ组和Ⅲ组T1,2时细胞胞浆Cty c浓度升高(P＜0.05);与Ⅱ组相比,Ⅲ组T2时细胞胞浆Cty c浓度升高(P＜0.05).不同时机异丙酚给药组细胞凋亡数目较M组明显减少.结论 异丙酚不同时机给药可减少线粒体Cty c释放到胞浆,抑制海马神经元凋亡,减轻缺氧复氧损伤,缺氧前给药效果较好.     

二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时细胞凋亡的影响

目的 探讨二氮嗪预先给药对大鼠心肌微血管内皮细胞缺氧复氧时细胞凋亡的影响.方法培养SD大鼠心肌微血管内皮细胞,以1×106个/ml的密度接种于96孔培养板(100 μl/孔)或培养皿(2 ml/皿),采用随机数字表法,将其随机分为4组(n=12),正常对照组(C组)不作任何处理,缺氧复氧组(H/R组)、二氮嗪预先给药组(DZ组)和二氮嗪预先给药+5-羟葵酸组(DZ+5-HD组)均进行缺氧2 h复氧2 h,DZ组和DZ+5-HD组在缺氧前2 h分别加入100 μmol/L二氮嗪和100 μmol/L二氮嗪+100 μmol/L线粒体ATP敏感性钾通道阻断剂5-羟葵酸.于复氧2 h时测定细胞活力和凋亡率.结果 与C组比较,H/R组细胞活力降低,细胞凋亡率升高(P＜0.01);与H/R组比较,DZ组细胞活力升高,细胞凋亡率降低(P＜0.05或0.01);5-羟葵酸可抑制二氮嗪预先给药导致的上述改变(P＜0.05或0.01).结论 二氮嗪预先给药可抑制大鼠心肌微血管内皮细胞凋亡,从而减轻缺氧复氧损伤,其机制与激活线粒体ATP敏感性钾通道有关.

Abstract：

Objective To investigate the effects of diazoxide pretreatment on apoptosis in rat myocardial microvascular endothelial cells exposed to hypoxia-reoxygenation (H/R) . Methods The SD rat myocardial microvascular endothelial cells were cultured. The cells were seeded in 96-well plates (100 μl/hole) or in 6 cm diameter dishes (2 ml/dish) with the density of 1×106/ml and randomly divided into 4 groups ( n = 12 each) : normal con trol group (group C), H/R group, diazoxide pretreatment group (group DZ) and diazoxide pretreatment + 5-hydroxydecanoate (5-HD, a mitochondrial ATP-sensitive potassium channel blocker) group (group DZ + 5-HD) .The cells were exposed to 2 h hypoxia followed by 2 h reoxygenation. Diazoxide 100 μmol/L and diazoxide 100 μmol/L + 5-HD 100 μmol/L were added to the culture medium 2 h before hypoxia in groups DZ and DZ + 5-HD respectively. The cell viability and apoptotic rate were detected at the end of reoxygenation. Results Compared with group C, the cell viability was significantly decreased, while the apoptotic rate increased in group H/R ( P ＜ 0.01) . Compared with group H/R, the cell viability was significantly increased, while the apoptotic rate decreased in group DZ (P ＜ 0.05 or 0.01) . 5-HD could inhibit diazoxide pretreatment-induced changes mentioned above ( P ＜ 0.05 or 0.01). Conclusion Diazoxide pretreatment can reduce H/R injury through inhibiting apoptosis in rat myocardial microvascular endothelial cells, and the mechanism is related to the activation of mitochondrial ATP-sensitive potassium channels.     

异丙酚对大鼠海马神经元缺氧复氧时线粒体膜通透性的影响

目的 探讨异丙酚对大鼠海马神经元缺氧复氧时线粒体膜通透性的影响.方法 原代培养胎鼠海马神经元,随机分为3组:对照组(C组)正常培养;模型组(M组)缺氧低糖培养2 h后复氧;异丙酚组(P组)缺氧低糖培养前加入异丙酚,终浓度20 μmol/L.各组于复氧后即刻、4、8、12和24 h(T1～5)时分别用噻唑蓝比色法测定神经元活力,流式细胞仪检测线粒体膜电位(MMP),于T5时采用激光共聚焦显微镜观察细胞膜与线粒体膜通透性.结果 与C组相比,M组T1-5时神经元活力和MMP降低(P＜0.05),T5时细胞膜与线粒体膜通透性增加;与M组相比,P组T1～4时神经元活力升高,T1～5时MMP增加(P＜0.05),T5时细胞膜与线粒体膜通透性降低,完整性改善.结论 异丙酚可通过提高MMP,改善细胞膜与线粒体膜通透性,增强神经元活力,从而减轻大鼠海马神经元缺氧复氧损伤.     

异丙酚对LPS诱导中性粒细胞Toll样受体2和Toll样受体4表达的影响

目的 探讨异丙酚对内毒素(LPS)诱导中性粒细胞Toll样受体2(TLR2)和TLR4表达的影响.方法 健康志愿者6名,年龄20～35岁,各采集外周静脉血样50 ml,加入20 U/ml肝素抗凝,分离提纯中性粒细胞,制备细胞悬液,然后随机分为6组,每组6皿:对照组(C组)不给予任何药物,置于37℃、5%CO_2培养箱中培养12 h;异丙酚脂质溶剂intralipid组(I组)、异丙酚组(P组)和LPS组(L组):分别加入intralipid(终浓度为5 μg/ml)、异丙酚(终浓度为5 μg/ml)或LPS(终浓度为1μg/ml),置于37℃、5%CO_2培养箱中孵育12 h;intralipid+LPS组(IL组)和异丙酚+LPS组(PL组)先分别加入intralipid(终浓度为5 μg/ml)或异丙酚(终浓度为5 μg/ml)后,置于37℃、5%CO_2培养箱中孵育20 min,然后加入LPS(终浓度为1μg/ml),置于37℃、5%CO_2培养箱孵育12 h.采用流式细胞仪测定中性粒细胞膜TLR2和TLR4的表达;采用荧光定量PGR检测中性粒细胞膜TLR2 mRNA和TLR4 mRNA的表达;采用ELISA法测定培养上清液TNF-α和IL-8的浓度.结果 与C组比较,I组和P组TLB2和TLR4的表达、1NF-α和IL-8L-8的浓度差异无统计学意义(P>0.05),L组和IL组TLR2和TLR4的表达上调,L组TNF-α和IL-8L-8的浓度升高,IL组IL-8浓度升高(P<0.05);与L组比较,IL组TLR2和TLR4的表达、TNF-α和IL-8L-8的浓度差异无统计学意义(P>0.05),PL组TLR2和TLR4的表达下调,TNF-α和IL-8L-8的浓度降低(P<0.05).各组TLR2 mRNA和TLR4 mRNA的表达差异无统计学意义(P>0.05).结论 异丙酚可下调LPS诱导的中性粒细胞TLR2和TLR4的表达,从而抑制炎性反应.     

异丙酚对脂多糖诱导大鼠腹腔巨噬细胞Toll样受体-4 mRNA表达的影响

目的 观察异丙酚对脂多糖（LPS）诱导大鼠腹腔巨噬细胞Toll样受体-4（TLR-4）mRNA表达的影响，探讨异丙酚抑制LPS诱导白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF—α）产生的机制。方法 雄性Wistar大鼠32只，处死后分离腹腔巨噬细胞，随机分为4组（n=8）：A组（阴性对照组）；B组LPS（终浓度为1μg／ml）／加入巨噬细胞中；C组LPS（终浓度为1μg／ml）＋异丙酚（终浓度为1μg／ml）加入巨噬细胞中；D组LPS（终浓度为1μg／ml）＋异丙酚（终浓度为5μg／ml）加入巨噬细胞中。细胞培养12h后。用ELISA方法检测培养上清液中IL-6、TNF-α的浓度，用RT-PCR方法检测TLR-4mRNA的表达水平。结果 与A组相比，B组IL-6、TNF-α和TLR-4m RNA水平均增加，C组IL-6、TLR-4m RNA水平升高（P〈0．01）；与B组相比，C组、D组IL-6、TNF-α和TLR-4m RNA水平降低（P〈0．05或〈0．01）。结论 异丙酚通过下调TLR-4m RNA的表达水平，从而一定程度上抑制了LPS诱导大鼠腹腔巨噬细胞，TNF-α和IL-6的产生。     

TLR2和TLR4在胰腺癌中的表达及其意义

目的探讨Toll样受体2(TLR2)和Toll样受体4(TLR4)在胰腺癌中的表达及其临床意义。方法采用实时荧光定量PCR检测30例新鲜胰腺癌及相应癌旁组织标本中TLR2及TLR4 mRNA的水平;应用免疫组化检测TLR2和TLR4蛋白在65例胰腺癌及相应38例癌旁组织中的表达;分析它们与临床病理特征的关系。应用Kaplan-Meier法分析TLR2和TLR4蛋白表达对患者生存时间的影响。结果PCR结果显示胰腺癌组织中的TLR2和TLR4 mRNA相对水平分别为0.84±0.17和0.81±0.10,显著高于癌旁组织的0.70±0.13和0.70±0.16(P0.05);免疫组化结果显示TLR2和TLR4在胰腺癌中的表达率分别为63.10%和69.2%,显著高于癌旁组织的34.20%和39.5%(P0.05),TLR2,TLR4的表达与患者性别、年龄、肿瘤部位及分化程度无关,而与肿瘤大小、淋巴结转移、血管侵犯及临床分期有关,上述4指标的分组间差异均有显著性。TLR2或TLR4阴性组患者的生存时间分别为18.4个月和19.4个月,显著长于TLR2或TLR4阳性组(均为12.4个月,P0.05)。结论TLR2及TLR4在胰腺癌中高表达;TLRs信号通路可能促进了胰腺癌的恶性进展。     

异丙酚预先给药对缺氧复氧鼠脑神经元的保护作用

目的：观察异丙酚预先给药对缺氧复氧鼠脑神经元神经细胞活力、一氧化氮(NO)产量、热休克蛋白(Hsp70)和Hsp70 mRNA表达的影响，探讨异丙酚有无脑保护作用及其机制。方法:培养12d的胎鼠大脑神经元，随机分为四组：Ⅰ组(正常对照组)；Ⅱ组(缺氧复氧组)；Ⅲ组(14μmol／L异丙酚预处理组)；Ⅳ组(56μmol／L异丙酚预处理组)。Ⅲ组和Ⅳ组于缺氧前1h分别换入含有14μmol／L和56μmol／L异丙酚的培养液，随后置入95％N2 5％CO2培养箱中缺氧30min。四组于复氧的1、2、4、6、24h(分别记为T1、T2、T4、L、L)分别用MTT(改良四甲基偶氮唑盐)细胞酶学分析法和硝酸还原酶法测定神经元神经细胞活力(用OD值表示)和NO产量，同时四组于复氧的1、3、8、24、48、72h分别用原位杂交法和免疫组化法观察Hsp70 mRNA及Hsp70阳性细胞百分率。结果与Ⅰ组相比，Ⅱ组T1-24各时点OD值降低；Ⅲ组、Ⅳ组T1、T2时点OD值升高；与Ⅱ组相比，Ⅲ组、Ⅳ组OD值均升高；Ⅲ组、Ⅳ组间差异无显著性。在T1、T2及T4时点，与Ⅰ组相比，Ⅱ组NO产量增高；与Ⅱ组相比，Ⅲ组、Ⅳ组NO产量降低；Ⅰ组与Ⅲ组、Ⅰ组与Ⅳ组、Ⅲ组与Ⅳ组间差异无显著性。与Ⅰ组相比，Ⅱ组Hsp70 mRNA及Hsp70阳性细胞百分率增高，且开始增高的时间分别为3h和8h，高峰时间分别为24h和48h；与Ⅰ组相比，Ⅲ组、Ⅳ组Hsp70 mRNA阳性细胞百分率高峰提前至8h；Ⅲ、Ⅳ组间差异无显著性；与Ⅱ组和Ⅲ组相比，Ⅳ组Hsp70高峰提前至24h，而Ⅲ组与Ⅱ组差异无显著性。结论:异丙酚预先给药可抑制缺氧复氧引发的神经细胞活力降低，减轻神经细胞的缺氧复氧性损伤，这可能与异丙酚可抑制NO产量增高及分别从转录和翻译两个水平诱导鼠脑神经元Hsp70 mRNA和Hsp70的表达高峰提前有关。     

盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时TLR4 mRNA表达的影响

目的 探讨盐酸戊乙奎醚预先给药对失血性休克大鼠急性肺损伤时Toll样受体4(TLR4)mRNA表达的影响.方法 健康SD大鼠40只,体重200～250 g,随机分为5组(n=8):假手术组(S组)、失血性休克致急性肺损伤组(ALI组)和低、中、高剂量盐酸戊乙奎醚预先给药组(P1～3组).S组仅行动静脉穿刺,不放血,ALI组股动脉放血至35～45 mm Hg制备急性肺损伤模型,P1～3组分别于放血前30 min股静脉注射盐酸戊乙奎醚0.3、1.0、3.0 mg/kg,随后制备急性肺损伤模型.各组复苏后4 h时处死大鼠取肺,称重后计算肺湿干重比,检测TLR4 mRNA和NF-κB p65蛋白的表达水平,观察病理学结果.结果 与S组比较,ALI组和P1组TLR4 mRNA、NF-κB p65蛋白表达水平及肺湿干重比升高(P＜0.05或0.01),P2.3组差异无统计学意义(P＞0.05);与ALI组比较,P2,3组TLR4 mRNA、NF-κB p65蛋白表达水平及肺湿干重比降低(P＜0.05或0.01);P2组和P3组上述指标比较差异无统计学意义(P＞0.05).P2,3组肺组织病理学损伤程度较ALI组明显减轻.结论 盐酸戊乙奎醚预先给药可通过抑制肺组织TLR4 mRNA表达上调,进而降低NF-κB活性,从而减轻失血性休克诱发大鼠的急性肺损伤.     

小檗碱预先给药对缺氧/复氧诱导人肾小管上皮细胞凋亡的影响

目的 探讨小檗碱预先给药对缺氧/复氧诱导人肾小管上皮细胞凋亡的影响.方法 培养人肾小管上皮细胞(HK-2),以1×106个/ml密度接种于培养皿(2 ml/皿)和96孔培养板(200μl/孔),采用随机数字表法,将其分为4组(n=30):正常对照组(C组)、小檗碱组(B组)、缺氧/复氧组(H/R组)和缺氧/复氧+小檗碱组(H/R+B组).B组和H/R+B组加入10 μmol/L小檗碱孵育2h,随后H/R组和H/R+B组进行缺氧24h复氧3h.测定细胞活力、细胞凋亡率、丙二醛(MDA)、超氧化物歧化酶(SOD)、caspase-3、活化caspase-3、细胞色素c、葡萄糖调节蛋白78(GRP78)和C/EBP同源蛋白(CHOP)表达,并计算Bax表达与Bcl-2表达的比值(Bax/Bcl-2).结果 与C组比较,H/R组和H/R+B组细胞活力、SOD活性和caspase-3表达降低,细胞凋亡率、MDA浓度和Bax/Bcl-2升高,活化caspase-3、细胞色素c、GRP78和CHOP表达上调(P＜0.05),B组上述指标差异无统计学意义(P＞0.05);与H/R组比较,H/R+B组细胞活力、SOD活性和caspase-3表达升高,细胞凋亡率、MDA浓度和Bax /Bcl-2降低,活化caspase-3、细胞色素c、GRP78和CHOP表达下调(P＜0.05).结论 小檗碱预先给药可抑制缺氧/复氧诱导人肾小管上皮细胞凋亡,其机制与抑制线粒体应激通路和内质网应激通路有关.     

异丙酚对缺氧复氧后大鼠海马神经元诱生型一氧化氮合酶表达的影响

目的 观察异丙酚对原代培养海马神经元缺氧复氧后诱生型一氧化氮合酶(iNOS)表达及存活率的影响。方法 原代培养SD大鼠海马神经元随机分为四组:正常对照组、缺氧组、缺氧 异丙酚4μg/ml组、缺氧 异丙酚12μg/ml组。MTT法测定体外缺氧4h复氧24h后海马神经元的存活率,免疫细胞化学法测定iNOS表达的程度。结果 与缺氧组比较,两异丙酚组缺氧复氧后海马神经元的存活率提高(P<0.05),iNOS表达的灰度值降低(P<0.05或0.01),iNOS阳性表达率降低,并呈剂量依赖性(P<0.01)。结论 异丙酚可降低大鼠海马神经元缺氧复氧后iNOS的表达,提高存活率。     

右美托咪啶对老年患者术后认知功能和围术期单核细胞Toll样受体2和Toll样受体4表达的影响

目的 探讨右美托咪啶对老年患者术后认知功能和围术期单核细胞Toll样受体2(TLR2)和TLR4表达的影响.方法 择期手术治疗的腰椎间盘突出症和腰椎骨折患者45例,年龄≥65岁,体重53～72 kg,ASA分级Ⅰ或Ⅱ级,采用随机数字表法,将其随机分为3组(n=15):对照组(Ⅰ组)和不同剂量右美托咪啶组(Ⅱ组和Ⅲ组).麻醉诱导结束后静脉输注右美托咪啶负荷剂量1.0μg/kg,输注时间15 min,然后以0.5μg/·kg-1·h-1(Ⅱ组)或1.0 μg·Jg-1·h-1(Ⅲ组)的速率静脉输注至术毕,Ⅰ组给予等容量生理盐水.于麻醉诱导前(T1)、手术开始1.5 h(T2)、术毕(T3)和术后24 h(T4)时取静脉血样,检测外周血单核细胞TLR2及TLR4的表达.分别于术前1d和术后7d时采用简易精神状态量表评分和韦氏成人记忆量表及智力量表评价认知功能,记录术后认知功能障碍的发生情况.结果 与Ⅰ组比较,Ⅱ组和Ⅲ组术后认知功能障碍发生率降低,T2～T4时单核细胞TLR2和TLR4表达下调(P＜0.05);与Ⅱ组比较,Ⅲ组术后认知功能障碍发生率降低,T～T4时单核细胞TLR2和TLR4表达下调(P＜0.05).结论 右美托咪啶可预防老年患者POCD的发生,其机制与抑制单核细胞TLR2和TLR4的表达有关.     

盐酸戊乙奎醚对内毒素性急性肺损伤大鼠肺组织Toll样受体4mRNA和Toll样受体2mRNA表达的影响

目的 探讨盐酸戊乙奎醚对内毒索性急性肺损伤大鼠肺组织Toll样受体4(TLR4)mRNA和Toll样受体2(TLR2)mRNA表达的影响.方法 健康SD大鼠60只,雌雄不拘,体重200～220g,采用随机数字表法,将大鼠随机分为5组(n=12),对照组(C组)、LPS组和低、中、高剂量盐酸戊乙奎醚组(P1组～P3组).C组腹腔注射生理盐水2ml;LPS组腹腔注射LPS 8mg/kg;P1组～P3组分别腹腔注射LPS 8 mg/kg和盐酸戊乙奎醚0.3、1.0和3.0 mg/kg.给药结束后6 h时开胸,心室取血,并取肺组织,采用ELISA法测定血清TNF-α和Ib-6的浓度,RT-PCR法测定肺组织TLR4 mRNA和TLR2 mRNA 的表达水平,并观察肺组织病理学结果.结果 与C组比较,LPS组、P1组～P3组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达均升高(P<0.05);与LPS组比较,P2组和P3组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达均降低(P<0.05),P1组上述指标差异无统计学意义(P>0.05);与P1组比较,P2组和P1组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达均降低(P<0.05);P2组和P3组血清TNF-α、IL-6浓度和肺组织TLR4 mRNA、TLR2 mRNA表达比较差异无统计学意义(P>0.05).P2组和P3组肺组织病理学损伤程度明显轻于LPS组.结论 盐酸戊乙奎醚可通过下调肺组织TLR4 mRNA和耵JR2 mRNA的表达,降低炎性反应,从而减轻大鼠内毒素性急性肺损伤.

Abstract：

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.     

盐酸戊乙奎醚对内毒素性急性肺损伤大鼠肺组织Toll样受体4mRNA和Toll样受体2mRNA表达的影响

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.     

盐酸戊乙奎醚对内毒素性急性肺损伤大鼠肺组织Toll样受体4mRNA和Toll样受体2mRNA表达的影响

Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.     

心肌肽素预先给药对大鼠海马神经元缺氧复氧损伤的影响

目的 研究心肌肽素(CMP)预先给药对原代培养大鼠海马神经元缺氧复氧损伤的影响 及相关机制。方法原代培养SD大鼠海马神经元,随机分为4组:对照组、缺氧复氧组、CMP 10 μg／ml 组、CMP 100 μg／ml组。对照组不实行任何处理,缺氧复氧组海马神经缺氧4 h复氧48 h,建立神经元缺 氧复氧损伤模型。CMP 10 μg／ml、CMP 100 μg／ml组缺氧前30 min于培养液中加入相应浓度的CMP,缺 氧复氧组加入相应的溶剂。采用四唑蓝比色法测定神经元的细胞活力,采用细胞免疫化学法测定神 经元Bcl-2蛋白表达水平。结果与对照组比较。缺氧复氧组、CMP10 μg／ml组、CMP100μg／ml组海马 神经元缺氧复氧后细胞活力降低,Bcl-2蛋白表达升高(P<0．05),CMP 100 μg／ml组细胞活力及Bcl-2 蛋白表达高于缺氧复氧组和CMP10μg／ml组(P<0．05),CMP 10μg/ml组与缺氧复氧组之间上述两指 标比较差异无统计学意义(P>0．05)。结论CMP100μg／ml可增强Bcl-2蛋白的表达,减轻缺氧复氧 后海马神经元损伤。     

右美托咪啶对脂多糖诱导大鼠外周血单核细胞Toll样受体4mRNA表达的影响

目的 探讨右美托咪啶对脂多糖(LPS)诱导大鼠外周血单核细胞Toll样受体4(TLR4)mRNA表达的影响.方法 健康雄性Wistar大鼠40只,取外周血分离培养单核细胞,采用随机数字表法,将其随机分为5组(n=8),A组:阴性对照;B组:单核细胞中加入LPS(终浓度为1μg/ml);C组:单核细胞中加入LPS(终浓度为1μg/ml)+右美托咪啶(终浓度为0.5 ng/ml);D组:单核细胞中加入LPS(终浓度为1μg/ml)+右美托咪啶(终浓度为5.0 ng/ml);E组:单核细胞中加入LPS(终浓度为1μg/ml)+右美托咪啶(终浓度为50.0 ng/ml).孵育24 h后,收集上清液,采用ELISA法测定TNF-α、IL-1β和IL-6的浓度,采用RT-PCR法测定TLR4 mRNA的表达.结果 与A组比较,B组TNF-α、IL-1p、IL-6的浓度升高,TLR4 mRNA表达上调(P＜0.01);与B组比较,C组、D组和E组TNF-α、IL-1β、IL-6的浓度降低,TLR4 mRNA表达下调(P＜0.05或0.01);与C组比较,D组和E组TNF-α、IL-1β、IL-6的浓度降低(P＜0.01),TLR4 mRNA表达差异无统计学意义(P＞0.05);D组和E组各指标比较差异无统计学意义(P＞0.05).结论 右美托咪啶可通过下调TLR4 mRNA表达,抑制TLR4的合成,从而抑制LPS诱导大鼠外周血单核细胞TNF-α、IL-1β和IL-6的生成与释放.

Abstract：

Objective To investigate the effects of different concentrations of dexmedetomidine on the expression of Toll-like receptor 4 (TLR4) mRNA in rat peripheral blood monocytes exposed to lipopolysaccharide ( LPS ). Methods Peripheral blood monocytes isolated from male Wistar rats were seeded in 24-well plate in RPMI 1640 liquid culture medium in CO2 incubator at 37 ℃ and 5% CO2 for 2 h, and were randomly divided into 5 groups ( n = 8 each): group A negative control; group B was exposed to LPS 1 μg/ml and C, D and E groups were exposed to LPS 1 μg/ml + dexmetomidine 0.5, 5.0 and 50.0 ng/ml respectively. The monocytes were then incubated for 24 h. The concentrations of TNF-α, IL-1β and IL-6 in the supernatant of the cultured monocytes were detected by ELISA. The expression of TLR4 mRNA in the monocytes was detected by RT-PCR.Results Exposure to LPS significantly increased the expression of TLR4 mRNA and the concentrations of TNF-α, IL-1β and IL -6 in group B as compared with group A ( P ＜ 0.01 ). Dexmedetomidine attenuated the LPS-induced increase in the expression of TLR 4 mRNA and the concentrations of TNF-α, IL-1β and IL-6 in a dose-dependent manner ( P ＜0.05or 0.01 ). Conclusion Dexmedetomidine can inhibit the synthesis of TLR4 and inhibit the secretion and dilivery of TNF-α, IL-1β and IL-6 by down-regulating the gene expression of TLR4 in rat peripheral blood monocytes exposed to LPS.