脂联素预先给药对内毒素血症大鼠肝损伤的影响

目的 评价脂联素预先给药对内毒素血症大鼠肝损伤的影响.方法 雄性Wistar大鼠80只,10～ 14周龄,体重250 ～ 300 g,采用随机数字表法,将其分为4组(n=20):对照组(C组)、内毒素血症组(LPS组)、脂联素预先给药组(APN组)和白细胞介素-10(IL-10)中和抗体干预组(IL-10抗体组).LPS组、APN组和IL-10抗体组腹腔注射脂多糖(LPS)20 mg/kg,APN组和IL-10抗体组于给予LPS前12 h腹腔注射基因重组脂联素6 mg/kg,IL-10抗体组于给予基因重组脂联素前30 min腹腔注射IL-10中和抗体3 mg.每组于注射LPS后2h时取10只大鼠,采集心脏全血,检测血清谷草转氨酶(AST)、谷丙转氨酶(ALT)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-10(IL-10)水平;每组于注射LPS后2h另取10只大鼠,取肝组织,检测丙二醛(MDA)含量、髓过氧化物酶(MPO)活性、IL-10 mRNA以及血红素氧化酶-1(HO-1) mRNA表达.结果 与C组比较,LPS组、APN组和IL-10抗体组血清AST、ALT及TNF-α、IL-1β、IL-10水平、肝组织MDA含量和MPO活性升高,肝组织HO-1 mRNA表达上调,LPS组和APN组IL-10 mRNA表达上调(P＜0.05);与LPS组比较,APN组和IL-10抗体组血清AST、ALT及TNF-α 、IL-1β水平、肝组织MDA含量和MPO活性降低,肝组织HO-l mRNA表达上调,APN组肝组织IL-10 mRNA表达上调(P＜0.05);与APN组比较,IL-10抗体组血清AST、ALT及TNF-α、IL-1β水平、肝组织MDA含量和MPO活性升高,血清IL-10浓度降低,肝组织HO-1 mRNA表达下调(P＜0.05).结论 脂联素预先给药可减轻内毒素血症大鼠肝损伤,其机制与诱导IL-10生成有关.     

极化液对内毒素血症大鼠肝损伤的影响

目的研究葡萄糖-胰岛素-钾(极化液,GIK)对内毒素血症大鼠肝损伤的影响。方法雄性SD大鼠60只,体重200~250g,随机分为三组:对照组,脂多糖组(LPS组,LPS 8mg/kg),GIK组(LPS 8mg/kg+GIK 4ml·kg~(-1)·h~(-1))。采用全自动生化仪检测腹腔注射LPS后3d和5d大鼠血清丙氨酸氨基转移酶(ALT)及天门冬氨酸氨基转移酶(AST)含量,ELISA法检测三组大鼠肝组织匀浆TNF-α含量,并行HE染色观察肝组织病理变化,TUNEL免疫荧光检测肝实质细胞凋亡情况。结果与注射后3d比较,注射后5dLPS组大鼠血清ALT、AST、肝组织匀浆TNF-α含量明显升高,而GIK组大鼠血清ALT、AST、肝组织匀浆TNF-α含量明显下降(P0.05)。与对照组比较,注射后3dLPS组和GIK组大鼠血清ALT、AST、注射后5dLPS组大鼠血清ALT、AST明显升高(P0.05),注射后3、5dLPS组和GIK组大鼠肝组织匀浆TNF-α含量、肝损伤等级评分、肝细胞凋亡指数明显升高(P0.05)。与LPS组比较,注射后3dGIK组大鼠肝组织匀浆TNF-α含量、肝细胞凋亡指数明显降低(P0.05),注射后5dGIK组大鼠血清ALT、AST、肝组织匀浆TNF-α含量、肝损伤等级评分、肝细胞凋亡指数明显降低(P0.05)。结论腹腔注射LPS可引起大鼠肝损伤,导致肝功能改变及肝细胞破坏;GIK可减轻LPS诱导的大鼠肝损伤。     

己酮可可碱对大鼠内毒素性急性肺损伤炎症反应的影响

目的探讨己酮可可碱对大鼠内毒素(LPS)诱导急性肺损伤(ALI)炎症反应的影响。方法腹腔注射0.01%LPS1mg/kg,16h后在机械通气下气管内滴注1.5mg/kg(0.5ml)LPS建立大鼠内毒素性ALI模型。24只大鼠随机分为生理盐水对照组(C组)、急性肺损伤组(LPS组)和己酮可可碱组(PTX组),每组8只。7h后处死大鼠。酶联免疫吸附法(ELISA)测定支气管肺泡灌洗液(BAL)中肿瘤坏死因子α(TNF-α)、白细胞介素-10(IL-10)的含量,测肺湿/干重比,并观察BAL白蛋白浓度。结果与LPS组相比,PTX组大鼠BAL中TNF-α浓度、肺湿/干重比以及BAL白蛋白浓度显著降低(P<0.05),而IL-10显著升高。结论己酮可可碱能显著抑制内毒素性急性肺损伤大鼠的炎症反应,具有一定的肺保护作用。     

异丙酚对内毒素诱导大鼠急性肺损伤的保护作用

目的 探讨不同剂量异丙酚对内毒素(LPS)诱导大鼠急性肺损伤(ALI)的保护作用。方法 股静脉注射内毒素(LPS)5mg/kg,建立大鼠ALI模型。24只健康雄性Wistar大鼠随机分为四组:对照组(C组,输注生理盐水),LPS对照组(L组,股静脉注射LPS后输注生理盐水),低剂量异丙酚治疗组(Lp1组,股静脉注射LPS后立即输注异丙酚5mg/kg,随后5mg·kg~(-1)·h~(-1)维持),高剂量异丙酚治疗组(Lp2组,股静脉注射LPS后立即输注异丙酚10mg/kg,随后10mg·kg~(-1)·h~(-1)维持),每组6只。于注射LPS后1、2、3、4h抽血并于4h时处死大鼠,酶联免疫吸附法(ELISA)测定血清和支气管肺泡灌洗液(BALF)中肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-10(IL-10)水平;测肺湿/干重比;并观察BALF中性粒细胞计数比、蛋白浓度。结果 L组大鼠肺湿/干重、BALF中性粒细胞计数比及蛋白浓度均明显增加(P<0.05),血清及BALF中TNF-α、IL-1β、IL-10水平显著性升高(P<0.01),而异丙酚治疗组的各项指标均较内毒素组减轻,大剂量作用更明显(P<0.01)。结论 异丙酚对内毒素诱导的大鼠急性肺损伤有保护作用,大剂量作用较明显。     

白细胞介素-10对内毒素血症大鼠肝损伤的影响

目的 评价白细胞介素-10(IL-10)对内毒素血症大鼠肝损伤的影响.方法 清洁级雄性Wistar大鼠100只,10 ～ 14周龄,体重250 ～ 300 g,采用随机数字表法,将其随机分为5组(n=20)∶对照组(C组)、内毒素血症组(LPS组)、1L-10组(IL-10组)、HO-1诱导剂钴原卟啉Ⅸ组(Co组)和HO-1抑制剂锌原卟啉Ⅸ组(Zn组).LPS组、IL-10组、Co组和Zn组腹腔注射脂多糖(LPS) 20 mg/kg,IL-10组、Co组和Zn组于给予LPS前3h腹腔注射重组人IL-10 1 μg,Co组和Zn组分别于给予重组人IL-10后2h腹腔注射钴原卟啉Ⅸ和锌原卟啉Ⅸ25 mg/kg.每组于注射LPS后24 h时取10只大鼠,采集心脏全血,测定血清谷草转氨酶(ALT)、谷丙转氨酶(AST)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)的水平;然后取肝组织,测定谷胱甘肽过氧化物酶(GSH-Px)和超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量和血红素氧合酶-1(HO-1)mRNA表达.每组取10只大鼠,记录给予LPS后72 h内大鼠生存情况.结果 与C组比较,LPS组、IL-10组、Co组和Zn组血清ALT、AST、TNF-α、IL-1β水平和肝组织MDA含量升高,肝组织GSH-Px、SOD活性和生存率降低(P＜0.05);与LPS组比较,IL-10组和Co组血清ALT、AST、TNF-α、IL-1β水平和肝组织MDA含量降低,肝组织GSH-Px、SOD活性、HO-1 mRNA表达和生存率升高(P＜0.05);与IL-10组比较,Zn组血清ALT、AST、TNF-α、IL-1β水平和肝组织MDA含量升高,肝组织GSH-Px、SOD活性和生存率降低(P＜0.05),Co组上述指标差异无统计学意义(P＞0.05).结论 IL-10可减轻内毒素血症大鼠肝损伤,其机制与诱导HO-1表达有关.     

七氟烷对急性肺损伤大鼠肺组织细胞凋亡的影响及其机制

目的 观察吸入七氟烷对脂多糖(lipopolysaccharides,LPS)致急性肺损伤(acute lung injury,ALI)大鼠肺组织细胞凋亡的影响,探讨可能机制.方法 18只雄性SD大鼠按照随机数字表分入3组,每组6只:①对照组;②模型组(LPS组);③七氟烷预处理组.LPS组气管内滴注LPS 5 mg/...     

右美托咪定对脂多糖诱导的脓毒症大鼠全身炎症反应和肺损伤的影响

目的观察脂多糖(LPS)对大鼠血浆和肺组织中的炎性因子和Toll样受体4(TLR4)表达的影响及右美托咪定的干预作用。方法雄性SD大鼠40只,250~300g,随机分为四组。Control组(n=10)生理盐水1ml·kg~(-1)·h~(-1)大鼠尾静脉泵注6h;DEX组(n=10)右美托咪定(负荷量6.5μg·kg~(-1)·h~(-1),10min;5μg·kg~(-1)·h~(-1)维持)大鼠尾静脉泵注6h;LPS组(n=10)经大鼠尾静脉注射7.5mg/kg的LPS后继续生理盐水泵注6h;LPS+DEX组(n=10)经大鼠尾静脉注射7.5mg/kg的LPS后泵注右美托咪定6h。以6h为实验终结点,并于此时行右心室取血和肺组织标本的制备。采用ELISA法检测血浆IL-1、IL-6、TNF-α浓度,Western blot法检测肺组织TLR4、髓样分化因子88(MyD88)、NF-κB蛋白含量;检测大鼠肺组织湿/干比(W/D);并用Murakami法评测肺损伤程度。结果与Control组比较,LPS组大鼠血浆IL-1、IL-6和TNF-α浓度明显升高,肺组织中的TLR4、MyD88、NF-κB蛋白含量明显升高(P0.01),肺W/D明显升高;LPS+DEX组上述指标差异均无统计学意义。Control组和DEX组未见明显肺损伤,LPS组肺间质水肿、炎性细胞浸润明显,LPS+DEX组肺损伤程度明显减轻(P0.01)。结论 LPS的刺激可以明显升高大鼠血浆中炎性因子以及肺组织中TLR4的表达水平,右美托咪定的干预可以减轻这一趋势,缓解大鼠的全身炎症和肺水肿的程度。     

血红素氧化酶-1介导脂联素在脓毒症肺损伤中的保护作用

目的 探讨血红素氧化酶-1(heme oxygenase,HO-1)介导脂联素(adiponectin,APN)在脓毒症肺损伤中的保护作用.方法 80只雄性Wistar大鼠采用随机数字表法分为4组(每组20只):对照组(C组)、脓毒症模型组(LPS组)、脂联素预处理组(APN组)和HO-1抑制剂锌原卟啉IX(zinc protoporphyrin IX,ZnPP IX)干预组(Zn组).各组分别于腹腔注射脂多糖(lipopolysaccharide,LPS) (20 mg/kg)或0.9％氯化钠溶液后6h,采用酶联免疫吸附法检测血清中肿瘤坏死因子(tumor necrosis factor-alpha,TNF-α)和白介素(interlukin-6,IL-6)的含量.采用分光光度法测定一氧化氮(nitric oxide,NO)浓度、肺组织丙二醛(malonaldehyde,MDA)和髓过氧化物酶(myeloperoxidase,MPO)活性变化.采用实时荧光定量聚合酶链反应(real-time PCR)法检测肺组织HO-1、诱导型一氧化氮合成酶(inducible nitric oxide synthase,iNOS)mRNA的表达水平.观察5d内大鼠生存情况,并绘制生存曲线. 结果 与C组比较,LPS组、APN组和Zn组血清TNF-α [(321.3±11.2)、(132.5±10.2)、(311.7±12.4)vs (52.1±1.4)] ng/L、IL-6[(2 229.26±210.25)、(1 134.14±11.24)、(2 028.27±167.04)vs(55.38±0.23)] ng/L、NO[(103±10)、(79±8)、(97±9)vs(48±3)] μmol/L和肺组织匀浆MDA[(16.209±0.151) 、(5.943±0.310)、(13.238±0.326)vs(3.081 ±0.017)] μmol/g、MPO[(12.42±0.46)、(6.77±0.14)、(10.45±0.21)vs(2.74±0.06)] U/g水平显著升高(P＜0.05),生存率显著下降(20％ vs 100％;70％ vs 100％;30％ vs 100％,P＜0.05);与LPS组比较,APN组血清TNF-α [(132.5±10.2)vs (321.3±11.2)] ng/L、IL-6 [(1 134.14±107.13)vs(2229.26±210.25)] ng/L、NO[(79±8)vs (103±10)] μmol/L和肺组织匀浆MDA[(5.943±0.310)vs (16.209±0.151)] μmol/g、MPO [(6.77±0.14)vs (12.42±0.46)] U/g水平显著降低(P＜0.05),肺组织HO-1 mRNA[(8.73±0.24)vs(1.54±0.03)]显著升高(P＜0.05),iNOS mRNA[(1.42±0.15)vs(2.01±0.10)]显著下降(P＜0.05),生存率显著增高(70％ vs 20％,P＜0.05);与APN组比较,Zn组血清TNF-α[(311.7±12.4)vs(132.5±10.2)] ng/L、IL-6[(2 028.27±167.04)vs(1 134.14±107.13)]ng/L、NO[(97±9)vs(79±8)] μmol/L和肺组织匀浆MDA[(13.238±0.326)vs(5.943±0.310)] μmol/g、MPO[(10.45±0.21)vs(6.77±0.14)] U/g水平显著升高(P＜0.05),肺组织HO-1 mRNA[(2.66±0.14)vs (8.73±0.24)]显著下降(P＜0.05),iNOS mRNA[(2.06±0.23)vs(1.42±0.15)]显著升高(P＜0.05),生存率显著下降(30％ vs 70％,P＜0.05). 结论 HO-1可能介导了APN在脓毒症性肺损伤中的保护作用.     

内毒素预处理对内毒素血症大鼠肺的作用及机制探讨

目的 观察内毒素预处理对内毒素血症大鼠肺的作用及其机制。方法 将雄性Wistar大鼠84只随机分为7组：生理盐水(NS)组,内毒素脂多糖(LPS)2h、4h、6h组和LPS预处理2h、4h、6h组,每组12只。LPS预处理各组大鼠经腹腔注射LPS0．25mg／kg,24h后再注射LPS0．5mg/kg,NS组和LPS各组在上述时间均给予等容量NS;第2次腹腔注射72h后,LPS各组和LPS预处理各组大鼠经静脉注入(静注)LPS 10mg／kg,NS组注射等量NS。NS组在静注NS后6h,LPS2h、4h、6h组和LPS预处理2h、4h、6h组在静注LPS后2、4、6h时各取6只大鼠取血,行血气分析;取左侧肺组织检测细胞间黏附分子-1(ICAM-1)mRNA及抑制性κB-α(IκB-α)蛋白表达;计数右肺支气管肺泡灌洗液(BALF)中白细胞数,测蛋白含量。上述7组另取6只大鼠,在上述相同时点取全肺,计算肺体指数,测定髓过氧化酶(MPO)。结果 LPS各组大鼠较NS组大鼠肺体指数、BALF中白细胞数和蛋白及肺组织MPO含量均增加,氧分压和HCO3^-下降;而LPS预处理各组大鼠上述各指标变化明显减轻。肺组织ICAM-1 mRNA在LPS2h、4h和6h组表达递增,而在LPS预处理各组表达显著减少;LPS2h组肺组织IκB-α蛋白表达较NS组减少,而LPS预处理2h组较LPS2h组表达增加。结论 内毒素预处理可防止内毒素血症时的肺损伤,可能与内毒素预处理使肺组织IκB-α蛋白生成增加和(或)消耗减少有关。     

中药丹参、生脉治疗全身炎症反应综合征的实验研究

目的探讨丹参、生脉治疗全身炎症反应综合征(SIRS)的作用及其机理。方法建立大鼠SIRS动物模型。40只Wistar大鼠随机分为4组，每组10只。对照组：生理盐水(5ml／kg) 脂多糖(LPS，1mg／kg)；丹参组：复方丹参注射液(5ml／kg) LPS(1mg／kg)；生脉组：生脉注射液(5ml／kg) LPS(1mg／kg)；联合用药组：复方丹参注射液(2，5ml／kg) 生脉注射液(2．5ml／kg) LPS(1mg／kg)。行血常规、血清TNF-α和IL-6水平检测，并行肝、肺及肾脏组织的病理组织学检查。结果丹参注射液及生脉注射液均能明显降低LPS腹腔注射后的血清TNF-α和IL-6水平，下调过高的炎症反应，并减轻肝、肺、肾等器官的损害．使SIRS大鼠48h存活率明显提高。结论本实验为丹参、生脉的抗SIRS临床应用提供了理论基础。     

Effects of Adiponectin on Mortality and Its Mechanism in a Sepsis Mouse Model

ABSTRACT

The mortality of sepsis is increasing and conventional therapies for it have no better therapeutic effects. We investigated the effects of adiponectin (APN) on mortality and high mobility group box 1 (HMGB1) in polymicrobial sepsis mouse models. Sepsis models were established by cecal ligation and puncture (CLP) in BALB/c mice. Animals were randomly divided into four groups including control group (C group), model group (CLP group), early APN treatment group (APN + CLP group), and late APN treatment group (CLP + APN group). Mice in each group were killed at 6, 12, 24, and 48 hr after CLP, respectively, to collect samples for determining the levels of serum interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), high mobility group protein-1 (HMGB1), and the expression of lung tissue HMGB1 mRNA. The survival curves in the four groups were drawn. The mortality rates were significantly lower in APN + CLP (30%) and CLP + APN (40%) groups than in CLP group (80%) seven days after CLP. Serum levels of cytokines (IL-6 and TNF-α) and HMGB1 were significantly reduced (p < .05) in APN + CLP and CLP + APN groups compared with those of CLP group. There was a significant correlation between serum HMGB1 and lung HMGB1 mRNA (r = 0.891). The levels of HMGB1 and HMGB1 mRNA were higher in CLP, APN + CLP, and CLP + APN groups than in C group (p < .01), but were lower in APN + CLP and CLP + APN groups than in CLP group (p < .01). APN can reduce the mortality rate and plays an anti-inflammatory role in polymicrobial sepsis mouse models through inhibiting HMGB1.     

血必净注射液对烫伤大鼠高迁移率族蛋白B1表达和急性肝损伤的影响

目的 了解血必净注射液对烫伤大鼠肝组织高迁移率族蛋白B1(HMGB1)表达及急性肝损伤的影响.方法 制作大鼠30%TBSA Ⅱ度烫伤模型,将78只大鼠按随机数字表法分为假伤组(18只)、烫伤组(30只)和血必净治疗组(30只),分别于伤后8、24、72 h处死取材.观察肝组织病理学变化;测定血清丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平;RT-PCR法检测肝组织HMGB1 mRNA表达,蛋白质印迹法及免疫组织化学法检测肝组织HMGB1蛋白相对表达量,结果 分别以积分吸光度比值、吸光度值表示.结果 光学显微镜下可见烫伤组大鼠肝组织有大量炎性细胞浸润,以伤后24 h较多;治疗组则明显减少.与假伤组比较,烫伤组大鼠血清ALT、AST水平显著增高(P<0.05或P<0.01),肝组织HMGB1基因和蛋白表达于伤后8～72 h显著增强(P<0.05或P<0.01).与烫伤组比较,治疗组大鼠血清ALT、AST水平有不同程度下降(P<0.05或P<0.01);伤后24、72 h肝组织HMGB1 mRNA表达分别为0.60±0.15、0.55±0.07,蛋白水平分别为163±13、160±16,显著低于烫伤组的0.75±0.12、0.78±0.11和200±13、175±14(P<0.05或P<0.01).结论 HMGB1 作为晚期炎性因子,参与了严重烫伤大鼠肝组织炎性反应的病理过程.血必净治疗可明显下调HMGB1表达,有助于减轻延迟复苏导致的急性肝损伤.     

Toll样受体4在高迁移率族蛋白B1影响小鼠调节性T细胞免疫功能中的作用

目的 探讨腹腔注射高迁移率族蛋白B1(HMGB1)后不同基因型小鼠天然调节性T细胞(CD4~+CD25~+Treg)免疫功能的改变及其受体作用机制.方法 分别给C3H/HeN和C3H/HeJ[分别为Toll样受体4(TLR4)野生型(TLR4~(+/+))和天然突变型(TLR4~(-/-)]小鼠腹腔注射不同剂量HMGB1(0、10、20μg/只),饲养48 h后采用免疫磁珠法分离小鼠脾脏CD4~+CD25~+Treg.体外培养12 h后采用流式细胞仪检测CD4~+CD25~+Treg表达细胞毒性T淋巴细胞相关抗原-4(CTLA-4)表达强度,并应用酶联免疫吸附试验(ELISA)检测CD4~+CD25~+Treg生成白细胞介素(IL)-10量.结果 与对照组比较,20 μg HMGB1攻击后C3H/HeN小鼠CD4~+CD25~+Treg表达CTLA-4水平显著下降(78.70±11.42与60.76±7.64,P<0.01),同时细胞牛成IL-10量也明显降低[(96.89±6.25)ng/L与(47.11±4.25)ng/L,P<0.01];但不同剂量HMGB1攻击可引起C3H/HeJ小鼠CD4~+CD25~+Treg表达CTLA-4明显上调和生成IL-10量不同程度地增加(P<0.01).结论 HMGB1攻击可显著影响CD4~+CD25~+Treg介导的免疫状态,TLR4在HMGB1诱导CD4~+CD25~+Treg免疫活性过程中发挥了重要负向调控作用.     

Immunotherapy improves immune homeostasis and increases survival rate of septic patients

Objective: To investigate the efficacy of immunotherapy on septic patients with Ulinastatin plus Thymosin-α1. Methods: Seventy postoperative septic patients were divided into two groups at random: the immunotherapy group (n=36) and the conventional therapy group (n=34). Patients in the immunotherapy group received intravenous Ulinastatin of 200 000 U, 3 times per day for 3 days, Ulinastatin of 100 000 U, 3 times per day for 4 days, and subcutaneous injection of Thymosin-α1 of 1.6 mg, twice per day for 3 days, then once per day for 4 days. While conventional therapies such as antibiotics and fluid resuscitation were undertaken in both groups. The expression levels of serum tumor necrosis factor-α (TNF-α), interleukin- 10 (IL-10), IgG, C3, T lymphocyte subsets, CD14+ monocyte human leukocyte antigen (locus) DR (HLA-DR) and patients' 28-day survival rate of the two groups were observed and evaluated. Results: The survival rate was significantly higher in the immunotherapy group (63.9%; 23/36) compared with the conventional therapy group (41.2%; 14/34). The serum TNF-α levels [(1.38±0.50) ng/ml in the immunotherapy group vs (1.88±0.53) ng/ml in the conventional group, P<0.05] and the serum IL- 10 levels [(217.52±15.71) ng/ml vs (101.53±16.57) ng/ml, P<0.05] were significantly different between the two groups. The serum IgG levels in the immunotherapy group [(17.65±6.81) g/L] were significantly higher than in the conventional group [(11.94±5.32) g/L]. There were also significant differences in the expression levels of CD4+ T lymphocyte (35%±13% in the immunotherapy group vs 21%±7% in the conventional group, P<0.05) and CD14+ monocyte HLA-DR (50%±5% in the former vs 35%±4% in the latter, P<0.05). Conclusions: Immunotherapy with Ulinastatin plus Thymosin-α1 can enhance the inflammatory response, improve the immune homeostasis, and increase the survival rate of septic patients.     

二氯甲烷对盲肠结扎穿孔诱导大鼠急性肺损伤的保护作用

目的 观察二氯甲烷对盲肠结扎穿孔模型大鼠急性肺损伤(acute lung injury,ALI)的保护作用.方法 40只SD大鼠按随机数字表法分为SHAM组、CLP组、MC组和DSF+MC组4组,每组10只.10 h后右侧颈总动脉放血处死.左肺行支气管肺泡灌洗,灌洗液进行白细胞计数,检测蛋白浓度和TNF-α及IL-10水平;右肺下叶观察大鼠肺组织病理变化,其他肺叶匀浆后检测丙二醛和髓过氧化物(myeloperoxidasedeficieney,MPO).另取40只大鼠随机分组处理同上,手术后分笼,观察其活动、刺激后反应;记录72 h生存率.结果 CLP组大鼠肺部炎症和损伤明显,与CLP组相比,MC组肺泡壁无增厚,肺泡内炎症细胞浸润减少,肺组织内TNF-α降低(55±16vs 164±36,P<0.01),IL-10增高(534.6±187.6 vs 75.2±26.1,P<0.01),丙二醛降低(7.8±1.7vs 30.4±2.2,P<0.01),术后72 h生存率显著增高(80%vs 0%,P<0.01).与MC组相比,MC+DSF组肺泡壁增厚,肺泡内炎症细胞浸润增多;肺组织内TNF-α增高(138±23 vs 55±16,P<0.01),IL-10降低(138.3±26.4vs 534.6±187.6,P<0.01),丙二醛升高(26.5±1.9 vs 7.8±1.7,P<0.05),术后72 h生存率显著增高(80% vs0%,P<0.01).结论 二氯甲烷对盲肠结扎穿孔所致的ALI具有明显的保护作用.     

不同免疫调节方式对烧伤脓毒症大鼠炎性反应的影响

Objective To investigate the effect of Thymosin and growth hormone(GH) on inflam-matory response in burn rats or burn rats with sepsis. Methods Sixty-four SD rats were randomly divided into normal control group (NC, without treatment), sepsis group (S, with injection of LPS), sepsis + Thy-mosin group (ST, with successive injection of Thymosin and LPS), sepsis + GH group [SGH, with succes-sive injection of recombinant human GH (rhGH) and LPS], burn group, burn + sepsis group (BS, with in-jection of LPS after burn), burn + sepsis + Thymosin group (BST, with successive injection of Thymosin and LPS after burn), burn + sepsis + GH (BSGH, with successive injection of rhGH and LPS after burn), with 8 rats in each group. Specimens of spleen tissues were harvested to determine HLA-DR in lymphocyte and e-valuate inflammatory cell infiltration (score). Specimens of peripheral blood were collected to determine Toll-like receptor 4 (TLR4) level in monocyte and serum level of TNF-α, IL-4, IL-6, IL-10. Results Compared with those in NC group, serum level of IL-10 in S group decreased obviously, while other indices increased obviously (P<0.01). The levels of HLA-DR and TLR4 and serum level of TNF-α were similar between SGH and ST groups (P>0.05). Compared with those in SGH group [(2.87±0.04) score, and IL-6 (0.0083±0.0018) μg/mg, IL-4 (0.0102±0.0021) μg/mg, IL-10 (0.0310±0.0027) μg/mg, re-spectively], degree of inflammatory cell infiltration (1.50±0.76) score and serum levels of IL-6, IL-4, IL-10 of rats in ST group decreased obviously (0.0064±0.0012, 0.0058±0.0024, 0.0230±0.0021 μg/mg, respectively, P<0.01). The levels of HLA-DR, TLR4 and inflammatory cell infiltration degree of spleen in B group were respectively higher than those in NC group and lower than those in BS group. Com-pared with those in NC group, serum levels of TNF-α, IL-6 in B group increased significantly, while IL-4, IL-10 showed an opposite tendency. There was no obvious difference between BST and BSGH groups in ser-um levels of HLA-DR and IL-6 (P>0.05). Compared with those in BST group, inflammatory cell infiltra-tion degree in spleen and the levels of TLR, TNF-α obviously decreased (P<0.01), while IL-4 and IL-10 levels increased in BSGH group (P<0.01). Conclusions Inhibitive effects between Thymosin and GH on extensive inflammatory reaction were similar with or without trauma, and GH has better effect as compared with Thymosin when with trauma.     

内毒素耐受对脓毒症大鼠肠道功能的保护作用

目的：动态观察内毒素耐受大鼠在内毒素血症时肠道炎症反应和免疫功能的变化,探讨内毒素耐受对内毒素血症大鼠肠道的保护作用及其机制。方法：选择健康成年雄性SD大鼠70只,随机分成3组：正常对照组（10只）、内毒素（LPS）组（30只）和内毒素耐受组（30只）。内毒素耐受组大鼠于实验第1日经腹腔注射LPS 0.25 mg/kg,24h后经腹腔注射LPS 0.5 mg/kg,LPS组在上述时间均给予等量生理盐水;实验第5日上述两组大鼠均腹腔注射大剂量LPS 10 mg/kg,于注射大剂量LPS前（0h）和注射后2、4、6、12h取大鼠肠组织制备肠黏膜上清液,用双抗体夹心法测定肠黏膜组织中分泌型免疫球蛋白A（sIgA）的含量,用免疫组化技术检测肠组织细胞间黏附分子-1（ICAM-1）的表达,并与正常对照组进行比较;光学显微镜下观察3组大鼠肠组织病理学变化,并进行病理学评分。结果：内毒素耐受组在给予小剂量LPS后没有出现体重下降。内毒素耐受组肠黏膜中slgA的含量较正常对照组明显升高,6 h最为明显（P〈0.05）,LPS组sIgA含量2 h明显升高（P〈0.05）,从6 h开始下降（P〈0.05）。内毒素耐受组给予大剂量LPS后肠黏膜ICAM-1的表达没有呈上升趋势,各时间点ICAM-1的表达与正常对照组比较差异有显著性,以6h ICAM-1的表达最低,而LPS组肠黏膜ICAM-1的表达随时间的延长逐渐升高,明显高于内毒素耐受组。肠组织病理学观察可见：LPS组肠黏膜水肿,绒毛脱落,并伴有大量炎性细胞浸润;内毒素耐受组上述表现较LPS组减轻。两组肠黏膜炎症损伤病理学评分[LPS组（3.89±0.45）分;内毒素耐受组（2.65±0.36）分]差异有显著性（P〈0.01）。结论：内毒素耐受可减轻内毒素血症导致的肠道黏膜屏障的损害,升高sIgA水平,减少ICAM-1表达,从而起到肠保护作用。     

右美托咪定预处理对大鼠心肌缺血/再灌注损伤时心肌组织高迁移率族蛋白B1的影响

目的 探讨右美托咪定(dexmedetomidine,Dex)预处理对大鼠心肌缺血/再灌注损伤(myocardial ischemia/reperfusion injury,MFRI)时心肌组织高迁移率族蛋白B1(high mobility group box-1 protein,HMGB1)表达量的影响. 方法 健康雄性SD大鼠52只,体重250～300 g,按随机数字表法分为4组(每组13只):假手术组(S组)、缺血/再灌注(ischemia/reperfusion,I/R)组(I/R组)、Dex+I/R组(D组)、Dex+育亨宾+I/R组(D/Y组).结扎左冠状动脉前降支30 min,恢复灌注120 min制备MI/RI模型.再灌120 min时采血ELISA法测定血清中IL-6、TNF-α浓度,随后处死大鼠摘取心脏,用2,3,5-氯化三苯基四氮唑(2,3,5-triphenyltetrazolium chloride,TTC)染色法测定心肌梗死面积,Western blot法测定心肌组织HMGB1蛋白的表达量. 结果 与S组比较,I/R组血清中IL-6、TNF-α含量[(336±41)、(636±25) ng/L]均显著增高(P＜0.05),心肌梗死面积明显增大(P＜0.05),心肌组织HMGB1蛋白表达量明显升高(P＜0.05);与豫组比较,D组血清中IL-6、TNF-α含量[(153±10)、(233±9) ng/L]均明显降低(P＜0.05),心肌梗死面积明显减少(P＜0.05),心肌组织HMGB1蛋白表达量显著降低(P＜0.05);与D组比较,D/Y组血清中IL-6、TNF-α含量[(230±20)、(386±32) ng/L]均显著增高(P＜0.05),心肌梗死面积明显增大(P＜0.05),心肌组织HMGB1蛋白表达量明显升高(P＜0.05). 结论 Dex预处理减轻MI/RI心肌组织HMGB1的表达量,减轻I/R损伤的炎症反应.Dex通过α2受体发挥保护作用.     

姜黄素抑制TLR4/HMGB1通路保护脂多糖诱导急性肺损伤的作用

目的探讨姜黄素对脂多糖(LPS)诱导急性肺损伤的保护作用及相关机制。方法将24只SD大鼠随机分为对照组、LPS组和LPS+姜黄素组,每组8只。检测肺损伤指标(氧分压、肺干湿重比、肺病理评分)、肺组织炎症反应程度[肿瘤坏死因子(TNF)-α、白介素(IL)-6、单核细胞趋化蛋白(MCP)-1以及肺组织中Toll样受体4(TLR4)、高迁移率族蛋白B1(HMGB1)表达水平]。结果LPS组氧分压显著低于对照组(P<0.05),而肺干湿重比、肺病理评分、TNF-α、IL-6、MCP-1、TLR4、HMGB1表达水平显著高于对照组(P<0.05);LPS+姜黄素组氧分压高于LPS组(P<0.05),肺干湿重比、肺病理评分、TNF-α、IL-6、MCP-1、TLR4、HMGB1表达水平显著低于LPS组(P<0.05)。结论姜黄素可能通过抑制TLR4/HMGB1通路,降低肺炎症反应程度,发挥保护LPS诱导急性肺损伤作用。