成年小鼠七氟醚麻醉手术后海马突触结合蛋白-Ⅰ表达的变化

目的 评价突触结合蛋白-Ⅰ(synaptotagmin-Ⅰ,syt-Ⅰ)在七氟醚麻醉的成年小鼠海马中的表达变化. 方法 成年雄性C57BL/6小鼠120只,16周龄,体重(21.2±2.2)g,采用随机数字表法分为4组(每组30只):吸氧+生理盐水+戊巴比妥钠组(C组)、吸氧+生理盐水+腹腔探查术+戊巴比妥钠组(S组)、七氟醚麻醉+腹腔探查术+生理盐水组(SS组)和七氟醚+腹腔探查术+地塞米松组(SD组).麻醉前1h,SD组腹腔注射地塞米松(2 mg/kg),余3组腹腔注射等量生理盐水.C组、S组1.5％戊巴比妥钠腹腔注射麻醉,SS组、SD组3.4％～3.6％七氟醚麻醉,S组、SS组、SD组行腹腔探查术.麻醉手术结束24 h后,水迷宫训练后测定各组的潜伏期、穿越平台次数、目标象限游泳时间.水迷宫测试结束后2h进行条件恐惧训练,条件恐惧训练结束24 h后进行场景恐惧和条件恐惧测试.行为学测试结束后取小鼠海马组织,Western blot检测syt-Ⅰ、IL-1β、S100A8含量,RT-PCR检测S100A8、Toll样受体4(toll-like receptor 4,TLR4)、IL-6、IL-1β、TNF-α,mRNA含量. 结果 与C组比较,S组和SS组第3、4、5天潜伏期延长、穿越平台次数减少、目标象限游泳时间缩短、场景恐惧时间下降(P均＜0.05);与SS组比较,SD组潜伏期缩短、穿越平台次数增加、目标象限游泳时间增加、场景恐惧时间增加(P均＜0.05).4组条件恐惧僵直反应差异无统计学意义(P＞0.05).与C组比较:S组和SS组IL-1β和S100A8蛋白及其mRNA表达增加,TNF-α mRNA表达增加(P＜0.05);SS组syt-Ⅰ及TLR4 mRNA含量降低,IL-1β和S100A8蛋白及其mRNA表达增加(P＜0.05). 结论 syt-Ⅰ表达下降参与了七氟醚麻醉手术后成年小鼠认知功能障碍.     

白细胞介素-33调节Th1/Th2细胞对小鼠肝热缺血再灌注损伤的保护作用

目的 研究白细胞介素(interleukin,IL)-33对小鼠肝热缺血再灌注(ischemia/reperfusion,I/R)损伤的保护作用,寻求缓解肝I/R损伤的有效方法.方法 采用小鼠肝脏热缺血再灌注损伤模型.首先检测小鼠肝脏I/R损伤时IL-33的mRNA和蛋白水平的变化.小鼠分对照组、模型组、重组IL-33干预组和抗ST2L抗体干预组,检测I/R损伤6h后天冬氨酸转氨酶(aspartateaminotransferase,AST)和丙氨酸转氨酶(alanine aminotransferase,ALT)水平、肝组织病理变化和血清肿瘤坏死因子(tumor necrosis factor,TNF)-α、干扰素(interferon,IFN)-γ、IL-4、IL-5、IL-13水平.结果 肝脏I/R损伤时,IL-33的mRNA和蛋白水平水平显著升高(t再灌注2h=-3.574,t再灌注6 h=-4.147; P＜0.05).重组IL-33干预后小鼠血清肝酶水平明显下降(tALT=4.592,tAST=3.471;P＜0.05),病理损伤程度减轻,IL-4、IL-5、IL-13水平升高(tIL-4=-4.995,tIL-5=-4.584,tIL-13=-4.431;P＜0.05),IFN-γ,水平下降(t=5.402,P＜0.05).抗ST2L抗体干预则有相反的作用.IL-33组和抗ST2L抗体组小鼠的血清TNF-α水平与模型组相比,差异无统计学意义(tTNF-α=0.261,P ＞0.05).结论 小鼠肝脏热缺血再灌注损伤时IL-33mRNA及血清蛋白表达水平显著增高,对发生I/R损伤的肝脏发挥保护作用,其机制可能与促进Th1/Th2平衡向后者偏移有关.     

不同生理阶段小鼠雌激素受体水平及IL-13、TNF-α含量的变化

目的观察小鼠在不同生理阶段雌激素及其受体及白细胞介素13(IL-13)和肿瘤坏死因子α(TNF-α)含量的变化,探讨雌激素水平对小鼠免疫功能的影响。方法清洁级健康雌性C57BL/6小鼠60只,其中2月龄小鼠(40只)随机分为发情间期组(对照组)、妊娠组,10月龄小鼠拟为自然绝经组,每组20只。3个月后进行实验,RT-PCR技术检测3组小鼠脾脏IL-13、TNF-αmRNA、ERαmRNA和ERβmRNA水平,ELISA检测血清雌二醇(E_2)水平,比较三组间各个指标差异。结果与对照组相比,妊娠组血E_2水平[(82.27±8.70)pmol/L vs.(55.23±5.11)pmol/L]和ERαmRNA水平[(3.93±1.54)vs.(2.70±1.17)]显著升高,自然绝经组E_2水平[(23.52±4.46)pmol/L vs.(55.23±5.11)pmol/L]和ERαmRNA水平[(1.15±0.71)vs.(2.70±1.17)]显著降低(P0.05);妊娠组小鼠脾脏IL-13、TNF-αmRNA水平显著降低,而绝经期小鼠脾脏IL-13、TNF-αmRNA水平显著升高,差异有统计学意义(P0.05);三组小鼠ERβmRNA水平无显著性差异(P0.05)。结论绝经组小鼠IL-13、TNF-α浓度水平较妊娠组及发情间期组升高,可能与其雌激素水平及ERα受体下降有关。     

脂联素基因重组慢病毒对早期糖尿病肾病小鼠肾脏的保护作用

目的 探讨静脉注射小鼠脂联素基因重组慢病毒对糖尿病肾病(DN)模型小鼠的肾脏保护作用及其机制.方法 40只C57BL/6小鼠按数字随机法分为正常对照组(NC组)、糖尿病肾病组(DN组)、阴性对照病毒(Lenti-IRES-EGFP)治疗组(DL组)、脂联素基因重组慢病毒( Lenti-Acdc-IRES-EGFP)治疗组(DA组),每组10只.应用链脲菌素(SFZ)结合高脂饮食诱导建立DN模型.重组慢病毒注射8周后,检测各组小鼠血浆脂联素水平、肾功能、尿白蛋白排泄率、肾组织病理改变.用免疫组织化学法原位检测肾组织增殖细胞核抗原( PCNA)表达.用Western印迹检测肾组织腺苷酸活化蛋白激酶α(AMPKα)、哺乳动物雷帕霉素靶蛋白( mTOR)水平.结果 成功构建脂联素超表达DN动物模型.与NC组比较,第12周末DA组小鼠肾质量指数(肾质量/体质量,KWI)、平均肾小球体积(MGV)、系膜面积比(FMA)、24h尿蛋白(UTP)均明显升高(P＜0.05),但低于DN组、DL组(P＜0.05).DA组肾组织PCNA阳性细胞数显著低于DN组、DL组(P＜0.01).与DN组、DL组相比,DA组小鼠肾组织AMPKα磷酸化水平显著升高(P＜0.01),mTOR磷酸化水平降低(P＜0.01).结论 脂联素通过激活AMPK信号通路,抑制mTOR信号活化,进而抑制早期DN时肾组织异常增殖.     

不同降温方式对脑外伤后伤灶区c-fos mRNA和神经生长因子表达的影响

目的 通过对大鼠重型脑挫裂伤后体温的干预,观察不同降温方式对脑外伤后伤灶区c-fos mRNA和神经生长因子(nerve growth factor,NGF)表达的影响,了解其表达强度是否与伤后不同降温方式对脑损伤的保护机制有关. 方法 健康成年SD大鼠336只,随机分为四组(每组84只),除假手术组外各组动物均按自由落体法造成重度脑挫裂伤;每组又按处死时间分为七个亚组(每亚组12只),分别检测c-fos mRNA(6只)与NGF(6只)的表达. 结果 ①伤后4h、8h、12 h与24h,全身亚低温组与局部亚低温组c-fos mRNA表达明显高于TBI组(P＜0.01),其他各时点无差异;TBI组在伤后4h、8h、12 h、24h与3d,c-fos mRNA表达均明显高于假手术组(P＜0.01).②伤后4h、8h,全身亚低温组与局部亚低温组NGF表达与TBI组无明显差异(P＞0.05);而在伤后12 h、24h、3d、5d与7d,两亚低温组NGF表达明显高于TBI组((P＜0.05或P＜0.01).③全身亚低温组与局部亚低温组在伤后各时间点c-fos mRNA和NGF表达无显著性差异(P＞0.05). 结论 脑外伤后亚低温可促进c-fos mRNA和NGF表达上调,全身亚低温与局部亚低温两种不同降温方式无明显差异.     

抗氧化剂对内毒素性休克大鼠α1-肾上腺素能受体mRNA表达的影响

目的评价抗氧化剂对内毒素性休克大鼠α1-肾上腺素能受体(α1-AR)mRNA表达的影响.方法雄性SD大鼠40只,随机分成5组(n=8)空白对照组(C组);内毒素休克组(LPS组,静脉注射LPS 15 mg·kg-1);异丙酚组(P组,注射LPS后1 h,静脉注射异丙酚10 mg·kg-1,继以10 mg·kg-1·h-1静脉持续泵注4 h);尿酸组(UA组,注射LPS后1 h,腹腔注射UA 200 mg·kg-1);N-乙酰5-甲氧基色胺组(MLT组,注射LPS后1 h,腹腔注射MLT 10 mg·kg-1).各组大鼠于注射LPS后6h处死,迅速取胸主动脉、下腔静脉、心脏、肝脏、肺脏、肾脏组织.提取各组织的总RNA,逆转录聚合酶链反应(RT-PCR)法检测各组织α1A-AR、α1B-AR和α1D-AR mRNA的表达.结果与C组比较,LPS组α1-AR三种亚型mRNA在胸主动脉、肝脏、肺脏、肾脏组织表达均下降(P＜0.05),α1A-AR、α1B-AR mRNA在下腔静脉、心脏组织表达均下降(P＜0.05).与LPS组比较,P组α1A-AR mRNA在肺脏和肾脏组织表达增加(P＜0.05),α1B-AR、α1D-AR mRNA在胸主动脉、肝脏、肺脏、肾脏组织表达增加(P＜0.05),α1B-AR mRNA在下腔静脉和心脏组织表达增加(P＜0.05);UA组α1A-AR mRNA在肾脏组织表达增加(P＜0.05),α1B-AR mRNA在肺脏以外各组织表达均增加(P＜0.05),α1D-AR mRNA在胸主动脉、肝脏、肺脏、肾脏组织表达增加(P＜0.05);但MLT组α1-AR三种亚型mRNA表达水平变化无统计学意义(P＞0.05).结论内毒素性休克大鼠α1-AR的基因表达普遍下调,抗氧化剂的抗休克作用机制与α1-AR基因表达上调有关.     

安定-氯胺酮麻醉对烧伤小鼠早期炎症反应的影响

目的研究安定-氯胺酮麻醉对烧伤小鼠腹腔巨噬细胞糖皮质激素受体(GR)及血清炎性细胞因子的影响,探讨其对创伤早期炎症反应的作用.方法 120只BALB/C健康雄性小鼠随机分为正常对照组、烧伤对照组、麻醉对照组、先处理组(麻醉后15 min烧伤)、后处理组(烧伤后15 min麻醉).采用安定-氯胺酮麻醉,小鼠烧伤模型为背部15%～20%Ⅲ度烧伤.于烧伤或麻醉后4 h颈椎脱臼法处死小鼠,收集全血及腹腔巨噬细胞.采用ELISA法测定血清肿瘤坏死因子(TNF-α)、IL-1β、IL-10浓度,采用Western blot技术测定腹腔巨噬细胞GR表达.取部分正常对照组和烧伤对照组腹腔巨噬细胞在体外与安定、氯胺酮共同培养,并测定腹腔巨噬细胞GR水平.结果与正常对照组比较,烧伤对照组血清TNF-α、IL-1β、IL-10浓度增高,先处理组血清IL-10浓度增高,后处理组血清IL-10浓度增高(P＜0.01);与烧伤对照组比较,先处理组和后处理组血清TNF-α、IL-1β、IL-10浓度降低(P＜0.01或0.05).与正常对照组比较,烧伤对照组、麻醉对照组和后处理组腹腔巨噬细胞GR表达水平降低(P＜0.05或0.01);与烧伤对照组比较,先处理组和后处理组腹腔巨噬细胞GR表达水平明显增高(P＜0.05或0.01);与先处理组比较,后处理组GR表达水平明显降低(P＜0.05).正常组或烧伤组腹腔巨细胞经安定-氯胺酮培养后GR表达差异无统计学意义(P＞0.05),正常组GR表达高于烧伤组(P＜0.01).结论安定-氯胺酮麻醉对小鼠烧伤后腹腔巨噬细胞GR表达水平下调以及炎性细胞因子的释放有一定抑制作用.     

地塞米松通过降低S100A8表达改善小鼠七氟醚麻醉手术后的认知功能障碍

目的 观察地塞米松对七氟醚麻醉手术后认知功能的改善及其作用机制. 方法 100只成年雄性C57BL/6小鼠,采用随机数字表法分为5组(每组20只):对照组(C组)、麻醉手术组(S组)、高剂量地塞米松治疗组(H组)、中剂量地塞米松治疗组(M组)和低剂量地塞米松治疗组(L组).C组不做处理,其余各组在麻醉后行腹腔探查术.H组、M组和L组分别给予20、2.0、0.2 mg/kg地塞米松,于术前1h经腹腔注射给药.术后24 h每组随机取10只小鼠海马组织,免疫组化检测海马小胶质细胞Iba1表达,RT-PCR检测S100A8、Toll样受体4(toll-like receptor 4,TLR4)、IL-6、IL-1β、TNF~ mRNA含量,Western blot检测S100A8蛋白.剩余小鼠进行5d水迷宫训练后,测定逃避潜伏期、穿越平台次数、目标象限游泳时间. 结果 与C组比较,S组小鼠术后逃避潜伏期[5d,(26.5±3.8)s]延长,穿越平台次数[(1.24±0.21)次]减少,目标象限游泳时间[(9.1±1.7)s]缩短(P＜0.05),小胶质细胞Iba1表达增加,S100A8(3.10±0.18)、TLR4(2.903±0.021)、IL-6(2.63±0.13)、IL-1β(3.733±0.160)、TNF-α(1.924±0.038) mRNA含量增加(P＜0.05).与S组比较,M组小鼠给药后逃避潜伏期缩短[5d,(17.8±2.3)s],穿越平台次数增加[(2.38±0.33)次],目标象限游泳时间延长[(14.5±2.0)s](P＜0.05);H组、M组小鼠小胶质细胞Iba1表达减少,S100A8(1.11±0.12、1.35±0.08)、TLR4(0.872±0.112、1.149±0.060)、IL-6(1.13±0.06、1.32±0.08)、IL-1β(0.755±0.059、0.941±0.133)、TNF-α(1.081 ±0.052、1.041±0.021)mRNA含量减少(P＜0.05). 结论 腹腔注射地塞米松(2.0 mg/kg),可改善小鼠七氟醚麻醉手术后认知功能,其机制可能与减少S100A8蛋白,抑制小胶质细胞活化,减少炎症因子表达有关.     

吸入不同浓度七氟醚麻醉对大鼠肺组织炎性反应的影响

目的 评价吸入不同浓度七氟醚麻醉对大鼠肺组织炎性反应的影响.方法 成年Wistar大鼠120只,雌雄不拘,体重200～250 g,随机分为4组:正常对照组(C组,n=12)、氧气组(O组,n=36)、1.5%七氟醚组(S1组,n=36)和3%七氟醚组(S2组,n=36).除C组外,其余各组根据吸入气体时间不同分为3个亚组(n=12),O组的3个亚组分别为4 h亚组、8 h亚组和10 h亚组;S1组和S2组的3个亚组分别为4 h亚组、8 h亚组和恢复亚组.C组仅吸入空气,O组吸入40% O2,S1组和S2组的4 h亚组和8 h亚组分别吸入40%O2+1.5%七氟醚或40%O2+3.0%七氟醚,恢复亚组吸入40%O2+1.5%或40%O2+3.0%七氟醚8 h后停止吸入七氟醚,然后吸入40%O2 2 h.于停止吸入气体时处死大鼠,测定支气管肺泡灌洗液(BALF)中TNF-α浓度;取肺组织,测定TNF-α mRNA表达和髓过氧化物酶(MPO)活性.结果 与C组比较,其他各组TNF-α浓度、TNF-α mRNA表达和MPO差异无统计学意义(P＞0.05);与O组的各亚组比较,S1组和S2组的各亚组上述指标差异无统计学意义(P＞0.05);S1组和S2组的各亚组间上述指标差异无统计学意义(P＞0.05).结论 吸入1.5%或3.0%七氟醚麻醉不诱发大鼠肺组织炎性反应.     

肿瘤坏死因子-α对急性肝衰竭小鼠肠黏膜通透性的影响及微生态制剂的保护作用

目的 研究急性肝衰竭小鼠生化指标及肠黏膜通透性改变,并探讨双歧杆菌乳杆菌三联活菌对肠黏膜通透性影响的机制及其对肝脏的保护作用.方法 采用数字表法将30只健康清洁级6～8周龄雄性BALB/c小鼠随机分为对照组、急性肝衰竭(ALF)组和微生态制剂双歧杆菌乳杆菌三联活菌干预组(干预组),每组各10只.干预组予双歧杆菌乳杆菌三联活菌(900 mg·kg-1·d-1)灌胃,对照组及ALF组予等量等渗盐水灌胃(9 mL·kg-1 ·d-1).2周后ALF组和干预组腹腔注射D-氨基半乳糖(3.0 g/kg),建立肝衰竭模型.注射后9h处死小鼠,留取血清、血浆观察生化指标.采用实时荧光定量PCR检测肝组织中肿瘤坏死因子-α(TNF-α)mRNA和回肠组织紧密连接蛋白(ZO)-1 mRNA的表达,采用免疫印迹检测回肠组织中ZO-1蛋白的表达.采用单因素方差分析或Kraskal-Wallis秩和检验比较各组间生化指标、TNF-α mRNA、ZO-1 mRNA以及ZO-1蛋白表达的差异,并采用Pearson检验分析ZO-1蛋白与血清TNF-α和血浆内毒素之间的相关性.结果 ALF组小鼠血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、TNF-α及血浆内毒素水平均明显高于对照组,差异有统计学意义(P＜0.01);干预组上述指标均较ALF组明显下降,差异有统计学意义(P＜0.01).ALF组小鼠肝组织TNF-α mRNA表达明显高于对照组,差异有统计学意义(Z =4.038,P＜0.01);干预组表达量较ALF组明显下降,差异有统计学意义(Z =3.780,P＜0.01).ALF组小鼠回肠ZO-1mRNA/蛋白表达均明显低于对照组,差异有统计学意义(P＜0.01);干预组表达量较ALF组明显增加,差异有统计学意义(P＜0.01).各组小鼠回肠ZO-1蛋白与血清TNF-α和血浆内毒素水平均呈负相关(r=-0.946和-0.919,P＜0.01).结论 TNF-α可能是导致肠黏膜通透性升高的原因之一.双歧杆菌乳杆菌三联活菌既能通过减轻内毒素对肝脏的损害而发挥保肝作用,又能上调回肠ZO-1蛋白的表达以改善肠黏膜通透性.     

Effects of metformin on expression of AMP - activated protein kinase in rat glomerular mesangial cells

ObjectiveTo observe the effects of metformin on expression of Adenosine 5’- monophosphate (AMP)-activated protein kinase (AMPK), nuclear factor-κB (NF-κB) and transforming growth factor β1 (TGF - β1) in cultured rat glomerular mesangial cells (MCs), and explore its reno - protective mechanisms. Methods MCs were cultured in the medium with normal glucose (group NG, 5.6 mmol/L), high glucose (group HG, 25mmol/L) and different concentrations of metformin (group M1, M2, M3). After 48 h exposure, the supernatants and MCs were collected. The expression of NF-κB and TGF-β1 mRNA was analyzed by real time-PCR. Total-AMPK, phospho-Thr-172 AMPK (p-AMPK), NF -κB p65 and TGF-β1 were visualized by Western blot. ResultsThe real time-PCR and Western blot result showed MCs could express AMPK, NF-κB and TGF-β1 mRNA and protein. After stimulated by HG, the levels of intracellular NF - κB and TGF - β1 expressions were significantly increased compared with group NG (P＜0.05); The levels of NF-κB and TGF-β1 were significantly decreased in group M1, M2 and group M3 compared with group HG in a dose-dependent manner. After stimulated by HG, the level of intracellular p-AMPK were down-regulated compared with group NG(all P＜0.05); The expression of p-AMPK increased with the rising of metformin concentration, presenting the opposite trend (P＜0.05), while the level of total-AMPK protein was unchanged with exposure to HG or different concentrations of metformin(P＞0.05). ConclusionMetformin can suppress the expression of NF- κB and TGF-β1 of glomerular MCs induced by HG via AMPK activation, which may partly contribute to its reno-protection.     

MicroRNA-148b influences high glucose-induced endoplasmic reticulum stress in rat mesangial cell by targeting AMPKα1

Objective To observe the expression of microRNA-148b (miR-148b) induced by high glucose in rat mesangial cells, and to explore its effect on its target gene AMP-activated protein kinase α1 (AMPKα1) and extracellular matrix excretion. Methods Rat mesangial cells were divided into 3 groups: normal glucose (NG, 5.5 mmol/L glucose) group, hypertonic (MA, 5.5 mmol/L glucose+ 19.5 mmol/L mannitol) group and high-glucose (HG, 25.0 mmol/L glucose) group. MiR-148b expression was detected by real time PCR. Then miR-148b inhibitor was transfected to rat mesangial cells. Their protein expressions of AMPKα1, glucose regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), fibronectin (FN) and collagen Ⅳ were detected by Western blotting. The expression of AMPKα1 mRNA was detected by real time PCR. The expression of collagen Ⅳ was also detected by immunofluorescence. Results Compared with NG group, HG group showed up-regulated miR-148b expression, down-regulated AMPKα1 mRNA and protein expressions, and up-regulated CHOP, GRP78, collagen Ⅳ and FN expressions (all P＜0.05). HG-induced mesangial cells with miR-148b inhibitor had up-regulated AMPKα1 mRNA and protein expressions, and down-regulated CHOP, GRP78, collagen Ⅳ, FN expressions as compared with HG-induced cells without miR-148b inhibitor (all P＜0.05). Conclusions HG can up-regulate miR-148b expression and down-regulate AMPKα1 expression in rat mesangial cells, then activate endoplasmic reticulum stress to induce extracellular matrix excretion. MiR-148b inhibitor up-regulates AMPKα1 expression, inhibits endoplasmic reticulum stress and reduces extracellular matrix excretion.     

异氟醚麻醉诱导大鼠脑海马神经炎症并激活Toll样受体2通路

目的 观察异氟醚麻醉与神经炎症的关系并探究其中的反应机制. 方法 将15只雌性SD大鼠用完全随机分组法分为3组(每组5只):对照组行热板实验,麻醉组行异氟醚麻醉、热板实验,空白组不做上述处理.热板实验后取大鼠大脑进行切片制作,免疫荧光双标检测观察Toll样受体2(toll like receptor 2,TLR2)在大脑海马区的表达,Westemblot检测海马区TNF-α、IL-1β、IL-6、单核细胞趋化蛋白-1(monocyte chemoattractant protein-1,MCP-1)以及TLR2表达水平. 结果 与对照组比较,麻醉组大鼠热缩足反射潜伏期(paw withdrawal latency,PWL)在异氟醚麻醉2h[(3 1±4)s比(19±3)s]和24 h[(25±4)s比(19±4)s]后显著增高(P＜0.05),而48 h时与对照组PWL比较,差异无统计学意义(P＞0.05).与对照组比较,麻醉组大鼠海马区TNF-α、IL-1β、IL-6、MCP-1以及TLR2表达水平均升高(P＜0.01). 结论 异氟醚麻醉促进海马区炎症因子表达,并激活TLR2信号通路.     

异氟醚、七氟醚吸入麻醉对鼠骨骼肌微循环白细胞活动的影响

目的 探讨异氟醚、七氟醚吸入麻醉对鼠骨骼肌微循环白细胞活动的影响。方法 选择SD雄性大鼠20只,随机分为两组,制备提睾肌微循环模型。吸入异氟醚、七氟醚麻醉后,分别记录吸入异氟醚、七氟醚1．5MAC3h内微循环、小动脉A1的直径和血流速度,微循环毛细血管后微静脉的白细胞滚动和粘附数量。结果 吸入异氟醚、七氟醚1．5MAC3h内HR,MAP,CVP和A1的直径和血流速度无明显改变（P＞0．05）。微循环毛细血管后微静脉的白细胞流动和粘附数量显著增加（P＜0．01）。结论 长时间吸入异氟醚、七氟醚后,可引起大鼠骨骼肌微循环毛细血管后微静脉的白细胞滚动和粘附数量显著增加。     

AMPK attenuates inflammation to reduce fibrosis induced by acute ischemia reperfusion injury in mice

Objective To observe the effect of adenosine monophosphate activated protein kinase (AMPK) on attenuating inflammation in fibrosis induced by acute ischemia reperfusion injury (IRI) in mice. Methods Forty eight male C57BL/6 mice were randomly divided into four groups: sham operation group (sham group), IRI group, AMPK inhibitor+IRI group (AMPK/IRI group) and normal saline+IRI group (NS/IRI group), 12 mice each group. The mice with renal IRI were occluded for 30 min through clipping bilateral renal pedicle, then released renal perfusion. Mice in sham group were performed the separation of renal pedicle without clipping. Mice in AMPK/IRI group and NS/IRI group were respectively intraperitoneal injected AMPK inhibitor and normal saline before IRI. At the 2 d after operation, 6 randomly-selected mice from each group were blooded by extraction eyeball to detect BUN and Scr. The renal histopathological changes were observed through HE staining. The mRNA expression of IL-1β, IL-6 and TNF-α was detected by real time PCR, and the level of AMPK phosphorylation was detected by Western blotting. At the 14 d after operation, Collagen 1 (COL1), α-SMA and fibronectin (FN) were detected by immunofluorescence and Western blotting in 6 remained mice from each group. The degree of kidney fibrosis was observed through sirus red staining. Results Compared with those in sham group, tubular interstitial damage was aggravated (P＜0.05), BUN and Scr were increased (P＜0.05), the mRNA expression of IL-1β, IL-6 and TNF-α was increased at the 2 d after operation (all P＜0.05), and the level of AMPK phosphorylation was activated in IRI group and NS/IRI group (all P＜0.05); the degree of kidney fibrosis and the expression of COL1, α-SMA and FN were increased obviously at the 14 d (all P＜0.05). Compared with those in IRI group, in AMPK/IRI group tubular interstitial damage was aggravated (P＜0.05), BUN and Scr were increased (all P＜0.05), the mRNA expression of IL-1β, IL-6 and TNF-α was increased at the 2 d (all P＜0.05), and the level of AMPK phosphorylation was decreased (P＜0.05). Moreover, the degree of kidney fibrosis and the expression of COL1, α-SMA and FN were increased obviously at the 14 d in AMPK/IRI group (all P＜0.05). Conclusions AMPK can ameliorate the acute renal ischemia reperfusion injury induce fibrosis in mice, and the mechanism may be related to the decrease of inflammatory reaction.     

异氟烷对发育期大鼠海马IL-1β mRNA、IL-6 mRNA和TNF-α mRNA表达的影响

目的 探讨异氟烷对发育期大鼠海马IL-1β mRNA、IL-6 mRNA和TNF-α mRNA表达的影响.方法 出生7 d的SD大鼠64只,随机分为2组(n=32):对照组(C组)和异氟烷麻醉组(I组).I组大鼠吸入1.5%异氟烷麻醉6 h,C组吸入空气.I组于麻醉前(T0,基础状态)、麻醉2 h(T1)、4 h(T2)、6 h(T3)、麻醉停止后4 h(L)、6 h(T5)、12 h(T6)、24 h(T7)时,C组在对应时点各取4只大鼠,测定海马IL-1β mRNA、IL-6 mRNA及TNF-α mRNA的表达水平.结果 C组各时点海马IL-1β mRNA、IL-6 mRNA和TNF-α mRNA比较差异无统计学意义(P＞0.05);与C组比较,I组海马T1～5 时IL-1βmRNA表达上调,T2,3 时IL-6 mRNA表达上调,T1～6时TNF-α mRNA表达上调(P＜ 0.05).结论 1.5%异氟烷麻醉可短暂上调发育期大鼠海马IL-1β mRNA、IL-6 mRNA和TNF-α mRNA的表达.     

Comparation of the effects of insulin-like growth factor-1 receptor inhibitor and insulin on renal interstitial macrophage infiltration in mice with type 2 diabetic kidney disease

Objective To compare the effect of insulin-like growth factor-1 receptor (IGF-1R) inhibitor and insulin on renal interstitial macrophage infiltration in mice with type 2 diabetic kidney disease (DKD) mice. Methods Twenty-four male C57BL/6 mice were selected. After 1 week of adaptive feeding, 6 rats were randomly selected as the control group. The other mice were intraperitoneally injected with streptozotocin (30 mg/kg) after 8 weeks of high-fat and high-sugar feeding. After 72 h, the type 2 diabetes mellitus (DM) models were successfully established if random blood glucose was greater than 16.7 mmol/L. After 8 weeks, if the proteinuria of DM mice increased, the DKD models were successful. DKD mice were divided into 3 groups by random number remainder method: DKD group (n=6), DKD+insulin group (insulin group, n=6, subcutaneous injection of 1-2 U/d insulin) and DKD+IGF-1R inhibitor (IGF-1R inhibitor group, n=6, administered with 30 mg?kg-1?d-1 IGF-1R inhibitor). They were continuously treated for 8 weeks. Random blood glucose was tested by glucometer. Blood and urine were collected, and biochemical indicators, such as serum creatinine, urea nitrogen and urine protein were measured by biochemical analyzer. Renal pathological changes were detected by hematoxylin-eosin staining (HE) and periodic acid-schiff staining (PAS). Suppressor of cytokine signaling 2 (SOCS2) mRNA and insulin-like growth factor-1 (IGF-1) mRNA were detected by in situ hybridization. The protein expressions of SOCS2, F4/80, Toll-like receptor 4 (TLR4) and CD68 were detected by immunohistochemistry. Results Compared with the control group, blood glucose, serum creatinine, serum urea nitrogen and urinary protein excretion rate were significantly higher in DKD mice (all P＜0.05), and CD68+ cells number, F4/80+ cells number and the expression of TLR4 in the tubulointerstitial of DKD mice were significantly higher (all P＜0.05). After intervention with insulin or IGF-1R inhibitor, serum creatinine, serum urea nitrogen and urinary protein excretion rate of DKD mice were significantly reduced (all P＜0.05). Insulin intervention could significantly reduce blood glucose in mice (P＜0.05), but had no significant effect on macrophages. Although IGF-1R inhibitor did not significantly reduce blood glucose, it could significantly reduce the number of CD68, F4/80 positive cells and the expression of TLR4 protein in renal interstitium of DKD mice (all P＜0.05). Compared with the DKD group, insulin intervention significantly reduced the expression of IGF-1 protein and mRNA (both P＜0.01), and increased the expression of SOCS2 mRNA and protein (both P＜0.01). And the expression of SOCS2 protein was correlated with the number of F4/80+ cells in insulin group (R2=0.8461, P=0.005). However, IGF-1R inhibitors had no significant effect on SOCS2 expression, but had better inhibition of macrophage infiltration. Conclusion IGF-1R inhibitor has a better inhibitory effect on DKD renal interstitial macrophage infiltration than insulin. The mechanism may be related to the fact that IGF-1R inhibitor does not up-regulate SOCS2 expression, whereas insulin up-regulates SOCS2 expression to activate some potential pathways.     

Effects of Sevoflurane and Isoflurane on Renal Function and on Possible Markers of Nephrotoxicity

Background: Low-flow sevoflurane anesthesia is associated with increasing circuit concentrations of compound A, which is nephrotoxic in rats, but the effect of compound A and low-flow sevoflurane anesthesia on renal function in humans is unclear. The authors compared the effects of high- and low-flow sevoflurane and isoflurane anesthesia on renal function and on several possible markers of nephrotoxicity in humans.

Methods: Forty-two patients without preexisting renal disease underwent either low-flow isoflurane (1 l/min, n = 14), low-flow sevoflurane (1 l/min, n = 14), or high-flow sevoflurane (6 l/min, n = 14) anesthesia for body-surface-area surgery scheduled to last at least 4 h. Twenty-four-hour urinary excretion of N-acetyl-[small beta, Greek]-glucosaminidase (NAG), [small beta, Greek]2-microglobulin, protein, glucose, blood urea nitrogen (BUN), and serum creatinine concentrations were measured before and after anesthesia.

Results: There were no differences in blood urea nitrogen, creatinine, and creatinine clearance among the three groups after anesthesia. Increased urinary N-acetyl-[small beta, Greek]-glucosaminidase excretions were seen in the low-flow and high-flow sevoflurane groups, but not in the low-flow isoflurane group (P < 0.01). Ten patients in the low-flow sevoflurane group had 24-h urinary excretion of protein that exceeded the normal ranges after anesthesia, but only one patient in the isoflurane and none in the high-flow sevoflurane groups had this.     

初生期异氟烷暴露对小鼠脑细胞发育、成年期行为、学习和记忆的影响

背景 异氟烷等吸入麻醉药广泛地应用于婴幼儿.异氟烷、咪达唑仑以及氧化亚氮暴露后,新生小鼠神经变性和神经认知功能损害的一些研究,已经引起越来越多人对儿科麻醉安全问题的关注.新生小鼠异氟烷暴露较长时间会触发可致神经认知功能损害的低血糖症.本研究即检测初生期异氟烷暴露和血糖对小鼠的脑细胞存活力、自主活动能力以及空间学习记忆能...     

异氟醚对大鼠海马高级糖基化终末产物受体表达的影响

目的 探讨异氟醚对大鼠海马高级糖基化终末产物受体(RAGE)表达的影响.方法 雄性老龄SD大鼠45只,月龄24月;雄性成年SD大鼠45只,月龄4月,分为老龄组和成年组(n=45),每组再分为3个亚组(n=15):老龄对照组(OC组)和成年对照组(AC组)吸入含30%氧气的空氧混合气体;老龄单次吸入异氟醚组(OS组)和成年单次吸入异氟醚组(AS组)吸入1.5%异氟醚2 h;老龄多次吸入异氟醚组(OR组)和成年多次吸入异氟醚组(AR组)吸入1.5%异氟醚3次,2 h/次,每天1次.吸入异氟醚后1 d各组随机取8只大鼠行Morris水迷宫实验测定认知功能,余大鼠处死取海马,采用RT-PCR法检测RAGE mRNA的表达水平,免疫组织化学法检测RAGE蛋白的表达水平.结果 与OC组比较,OS组和OR组认知功能减退,海马RAGE mRNA及其蛋白的表达上调(P＜0.05);与AC组比较,AS组和AR组认知功能减退,AR组海马RAGE mRNA及其蛋白的表达上调(P＜0.05);与OS组比较,OR组认知功能减退,海马RAGE mRNA表达上调(P＜0.05);与AS组比较,AR组认知功能减退,海马RAGE mRNA及其蛋白的表达上调(P＜0.05).结论 异氟醚可导致老龄和成年大鼠认知功能降低,尤其对老龄大鼠影响明显,可能与其上调海马RAGE表达有关.