c-Jun氨基末端激酶通路在内毒素休克大鼠生物喋呤诱生中的信号机制

目的 观察c-Jun氨基末端激酶(JNK)信号通路抑制剂——curcumin对内毒素休克大鼠生物喋呤(BH4)及一氧化氮(NO)表达的影响，并探讨JNK途径在生物喋呤诱生中的信号转导机制。方法建立内毒素休克模型，60只大鼠随机分为正常对照组(n=8)、内毒素休克组(n=32)和curcumin拮抗组(n=20)。采用逆转录多聚酶链式反应测定肝、肺、肾组织三磷酸鸟苷环水解酶Ⅰ(GTP-CHI)与诱生型一氧化氮合酶(iNOS)mRNA表达水平，分别采用反相高效液相色谱法和Griess比色法测定血浆与组织中BH4、NO水平的变化。结果与正常对照组相比，内毒素使动物肝、肺、肾组织GTP-CHI基因表达和BH4水平明显升高，至伤后24h仍持续于较高水平；与之相应，组织iNOS基因表达和NO水平亦明显升高。curcumin处理可明显下调肝、肺、肾组织GTP-CHI mRNA表达水平，并且肝组织6～24h时、肺组织12h时BH4水平显著降低；各组织iNOS mRNA表达及NO水平亦显著降低。结论抑制JNK信号通路能明显抑制内毒素休克鼠多器官生物喋呤／NO系统的表达，JNK途径参与了生物喋呤诱生的信号转导过程。     

内毒素休克大鼠生物喋呤改变与多器官损害的关系

目的探讨内毒素休克动物肝、肺、肾等重要器官生物喋呤(BH4)变化规律及其与多器官损害的关系。方法采用内毒素休克模型。104只大鼠随机分为正常对照组(n=8)、内毒素休克组(n=48)和BH4合成抑制剂2,4二胺6羟基嘧啶(DAHP)拮抗组(n=48)。肝、肺、肾组织BH4含量用反相高效液相色谱法测定,组织三磷酸鸟苷环水解酶I(GTPCHI,BH4合成的首要限速酶)基因表达采用逆转录PCR方法检测。结果内毒素攻击后0.5～2h动物肝、肺、肾组织BH4含量即开始升高(P<0.05),6～12h达峰值(P<0.01),此后逐渐降低;同时,组织GTPCHImRNA表达均明显高于正常对照组(P<0.05)。给予DAHP处理后,2～24h肝、肺、肾组织BH4水平均显著低于内毒素休克组(P<0.05或0.01),肝、肾组织GTPCHImRNA表达各时相点亦明显下调,肺组织12h后降低至正常对照范围。反映肝、肾功能指标和肺组织髓过氧化物酶活性均有不同程度地下降(P<0.05或0.01)。结论生物喋呤参与了内毒素诱发组织损害的病理生理过程,抑制BH4合成有助于减轻脓毒性休克所致过度炎症反应和多器官功能损害。     

生物喋呤对内毒素休克大鼠肺组织核因子-κB活化的影响及意义

目的　探讨生物喋呤对内毒素休克大鼠肺组织核转录因子 κB (NF κB)通路活化的影响及其在急性肺损伤中的意义。方法　采用内毒素休克模型 ,10 4只大鼠分为正常对照组 (n =8)、内毒素休克组(n =4 8)、 2 ,4 二胺 6 羟基嘧啶 (2 ,4 diamino 6 hydroxy apyrimidine ,DAHP)拮抗组 (n =4 8)。采用逆转录多聚酶链式反应测定肺组织三磷酸鸟苷环水解酶I (GTP CHI)mRNA的表达水平 ,采用凝胶阻滞电泳分析技术测定NF κB的DNA结合活性 ,同时测定肺组织髓过氧化物酶 (MPO)活性。结果　与正常对照组相比 ,内毒素攻击可导致动物肺组织GTP CHImRNA表达明显上调 ,6h后一直维持在较高水平 ;同时 ,NF κB活性于 0 5h迅速增加 ;且持续至伤后 4 8h ,肺组织MPO活性各时间点均显著升高 (P <0 .0 1) ,6h达峰值。采用生物喋呤合成抑制剂DAHP处理后 ,肺组织GTP CHImRNA表达早期改变不明显 ,但 12h后明显受抑 ;MF κB活性 0 5、 2 4、 4 8h明显降低 ;肺组织MPO活性 0 5、 6、 2 4、 4 8h均显著低于内毒素休克组 (P <0 0 5 )。结论　生物喋呤参与了内毒素休克动物肺组织NF κB信号通路的活化过程 ,抑制生物喋呤能降低NF κB的DNA结合活性 ,并减轻急性肺损伤     

休克期切痂对烫伤大鼠肝HMGB1表达及肝功能的影响

目的　探讨休克期切痂对烫伤大鼠肝组织高迁移率族蛋白B1(HMGBl)表达及肝脏功能影响。方法　 30 %Ⅲ度烫伤Wistar大鼠随机分为 2 4h和 72h切痂植皮组。RT PCR和免疫组化染色法检测肝脏HMGBlmRNA/蛋白表达 ,同时检测血浆AST、ALT含量。结果　大鼠烫伤后肝组织HMGB1mRNA表达量增加 1～ 2 5倍 ,2 4h切痂组伤后 8d其水平较烫伤对照和 72h切痂组显著减少 (P <0 0 5 )。烫伤后大鼠肝细胞和枯否细胞HMGB1表达阳性率较正常大鼠均显著升高 (P <0 0 1) ,2 4h切痂组 4、 8dHMGB1表达阳性细胞数较烫伤对照和 72h切痂组均显著减少 (P <0 0 1)。同时 ,烫伤大鼠血浆AST和ALT水平升高 (P<0 0 5 ) ,而 2 4h切痂组伤后 4、 8d较烫伤对照组和 72h切痂组显著降低 (P <0 0 1)。结论　烫伤大鼠休克期切痂能够下调肝组织HMGB1表达 ,局部HMGB1诱生参与了肝损伤的病理生理过程。     

血红素氧合酶-1对内毒素休克大鼠肺组织的保护功能

目的　探讨血红素氧合酶是否对内毒素休克大鼠的肺组织有保护作用。方法　将 4 0只健康清洁级Sprague Dawley大鼠随机分为对照组 (C组 ) ,内毒素休克组 (ES组 ) ,内毒素休克 +ZnPP IX组 (EZ组 ) ,ZnPP IX组 (Z组 )。检测肺组织含水率、MDA含量和SOD活性及HO 1mRNA、HO 2mRNA、HO 1和HO 2蛋白水平的变化。结果　①ES组大鼠肺组织含水率及MDA明显低于EZ组 (P值均小于 0 0 5 ) ,但其SOD明显高于EZ组 (P <0 0 5 ) ;②EZ组大鼠其肺组织组织内HO 1mRNA表达水平及HO 1蛋白水平明显低于ES组 (P值均小于 0 0 5 ) ;而各组间肺组织组织内HO 2mRNA表达水平及HO 2蛋白水平均无显著性差异(P值均大于 0 0 5 )。结论　内毒素休克大鼠其肺组织内的HO 1可能对其发挥着保护作用     

Alpha-黑素细胞刺激素对内毒素血症小鼠肝肺组织表达高迁移率族蛋白1的抑制作用

目的 研究Alpha-黑素细胞刺激素( α-MSH)对内毒素血症小鼠肝、肺组织中表达高迁移率族蛋白1(HMGB1)的调控作用.方法 腹腔内注射(ip)LPS(25 g/kg)和D-Gal(100 mg/kg)建立内毒素血症小鼠模型,分别在LPS刺激小鼠1、2、3 h后腹腔注射α-MSH(2.5 mg/kg),24 h后处死小鼠,取小鼠肺、脑、脾、肝、肾组织,应用RT-PCR和Western blot的方法,从基因、蛋白两个水平检测α-MSH对HMGB1表达的调节作用.结果 HMGB1 mRNA及蛋白在内毒素血症小鼠肺、脑、脾、肝、肾中均有表达,但在肝、肺中的表达量高于其他组织;于LPS刺激小鼠3 h之内腹腔注射α-MSH,能显著下调HMGB1 mRNA及蛋白的表达,并且1 h之内给药效果最理想.结论 α-MSH能有效抑制内毒素血症小鼠肝、肺组织中HMGB1的表达.     

杀菌/通透性增加蛋白对脓毒症大鼠组织TNF-α表达的影响及意义

目的　探讨杀菌 /通透性增加蛋白 (BPI)对脓毒症动物肝、肺肾组织肿瘤坏死因子 α (TNF -α)表达及多器官功能障碍的影响与意义。方法　采用大鼠盲肠结扎穿孔 (CLP)造成脓毒症模型。动物分为正常对照组 (10只 )、CLP组 (2 0只 )及BPI治疗组 (2 0只 )。CLP后 12h和 2 4h处死动物 ,分别检测肝、肺、肾组织TNF αmRNA表达、TNF α蛋白水平和器官功能指标。结果　CLP后 12h动物肝、肺、肾组织TNF αmRNA表达均显著增强 ,分别为伤前基础值的 2 4 6倍 (P <0 0 5 )、 2 86倍 (P <0 0 1)、 2 2 7倍 (P<0 0 1) ,同时各组织TNF α水平迅速升高 (P <0 0 1)。重组BPI治疗可不同程度地抑制肝、肺、肾组织TNF -αmRNA表达上调 (P <0 0 5 ) ,12h各组织TNF α水平显著降低 (P <0 0 5 ) ,均恢复至伤前正常范围。此外 ,BPI组动物血清丙氨酸转胺酶、肌酐水平在CLP后 12h显著低于未治疗组 ,CLP后 2 4h肺组织髓过氧化物酶活性也明显下降 (P <0 0 1)。结论　严重腹腔感染早期应用重组BPI治疗有助于下调TNF α的过量表达 ,从而抑制机体过度炎症反应 ,防止多器官功能障碍的发生与发展     

生物喋呤对烫伤脓毒症大鼠一氧化氮合成的影响

目的：探讨生物喋呤BH4在金黄色葡萄球菌（简称金葡菌）脓毒症中的生物学效应，阐明BH4对一氧化氮（NO）诱生的调控作用。方法：76只Wistar大鼠随机分为正常对照组（n=10)、烫伤对照组（n=10)、烫伤后金葡菌感染组（n=40)和羟基嘧啶（DAHP）拮抗组（n=16)。无菌留取动物肝、肺组织采用RT－PCR方法检测三磷酸鸟苷环水解酶I（GTP－CHI）、诱生型一氧化氮合酶（iNOS)基因表达，同时测定组织中BH4和NO的水平。结果：烫伤后金葡菌感染组动物肝、肺组织中GTP－CHI基因表达明显上调，BH4产生显著增加，iNOS mRNA表达和NO的水平亦明显升高，DAHP组GTP－CHI基因表达上调和BH4合成NO的产生亦明显下降。结论：烫伤后金葡菌感染可诱导体内BH4的合成，BH4在基因和蛋白水平调控着iNOS所介导的NO大量生成，从而对金葡菌脓毒症的病理过程起促进作用。     

严重腹腔感染大鼠组织Toll样受体2/4基因表达及其调节机制

目的 :研究腹腔感染后主要脏器 Toll样受体 (TL R) 2 / 4 m RNA表达的变化规律及组织分布特点 ,并对其诱生机制进行初步探讨。方法 :采用大鼠盲肠结扎穿孔 (CL P)造成严重脓毒症模型。动物分为正常对照组(10只 )、假手术组 (10只 )、CL P组 (6 0只 )及杀菌 /通透性增加蛋白 (BPI)治疗组 (2 0只 )。分别检测肝、肺、肾、小肠组织 TL R2 / 4 m RNA表达及血浆肿瘤坏死因子α(TNFα)和白介素 10 (IL 10 )水平。结果 :CL P后 2 h肝、肺、肾及小肠组织中 TL R2 / 4 m RNA表达开始增高 ,伤后 6～ 12 h各组织 TL R2 / 4 m RNA表达迅速达峰值。 TL R4 m RNA表达于伤后 2 4～ 4 8h开始下降 ,CL P后 72 h趋于伤前范围甚至阴性 ,而 TL R2 m RNA表达则持续增高至 72 h。早期给予 BPI治疗后 ,动物 12～ 2 4 h肝、肺、肾及小肠组织 TL R2 m RNA水平均显著降低(P<0 .0 5或 P<0 .0 1) ,同时 CL P后 12 h各组织 TL R4 m RNA表达亦不同程度下调 (P<0 .0 5或 P<0 .0 1)。BPI治疗组伤后 12 h血浆 TNFα水平显著降低 (P<0 .0 5 ) ,恢复至伤前正常范围 ,但伤后 2 4 h血浆 IL 10水平显著升高 (P<0 .0 1)。结论 :严重腹腔感染可迅速上调体内多器官 TL R2 / 4的基因表达 ,诱导促炎细胞因子产生 ,TL Rs表达与内毒素的直接刺激作用密     

选择性消化道脱污染对烫伤大鼠组织白细胞介素－18mRNA表达的影响

目的：探讨选择性消化道脱污染对烫伤大鼠组织白细胞介素－18mRNA表达的影响。方法：采用大鼠30％体表面积Ⅲ度烫伤模型,实验动物随机分为正常对照组,烫伤延迟复苏组,选择性消化道脱污染（SDD）预防组,肠,肺,肝,肾等组织IL－18mRNA含量采用逆转录聚合酶链式反应检测。结果：烫伤后体循环内毒素水平显著升高,8h,24h达高峰（P＜0．01）,给予SDD预治疗可显著降低内毒素峰值（P＜0．05）,另一方面,烫伤后2h肠,肺,肝,肾等组织IL－18mRNA表达较伤前即有显著升高,烫伤后8h达高峰（P＜0．01）,且一直持续至伤后24h,给予SDD预治疗后可不同程度抑制IL－18mRNA的表达（P＜0．05－0．01）,续集：肠,肺,肝,肾等组织IL－18mRNA表达在烫伤早期即显著增多,并呈一逐渐升高的趋势,创伤后内毒素血症对机体多种组织IL－18mRNA基因表达具有重要影响,SDD预治疗后可不同程度抑制IL－18mRNA的表达。     

Effect of 2,4-diamino-6-hydroxy-pyrimidine on postburn Staphylococcus aureus sepsis in rats

OBJECTIVE: Guanosine triphosphate-cyclohydrolase I (GTP-CHI) is the first and rate-limiting enzyme for the de novo biosynthesis of biopterin. The objective of present study was to observe the effect of 2,4-diamino-6-hydroxy-pyrimidine (DAHP), an inhibitor of GTP-CHI, on the development of postburn Staphylococcus aureus sepsis. DESIGN: A prospective, controlled animal study. SETTING: A research laboratory in a hospital. SUBJECTS: Male Wistar rats. INTERVENTIONS: Fifty-six male Wistar rats were randomly divided into four groups as follows: normal control group (n = 10), scald control group (n = 10), postburn sepsis group (n = 20), and DAHP treatment group (n = 16). In the scald control group, rats were subjected to a 20% total body surface area third-degree scald injury and then were killed at 24 hrs. In the postburn sepsis group (n = 20), rats were inflicted with 20% total body surface area third-degree scald followed by Staphylococcus aureus challenge, and they were further divided into 2- and 6-hr groups. In the DAHP treatment group (n = 16), animals were intraperitoneally injected with a dose of 1 g/kg DAHP before Staphylococcus aureus challenge and then were further divided into 2- and 6-hr groups. Tissue samples from liver, kidneys, lungs, and heart were collected to determine GTP-CHI, inducible nitric oxide synthase, and tumor necrosis factor-alpha messenger RNA expression. Meanwhile, biopterin and nitric oxide concentrations in these tissues were also measured. MEASUREMENTS AND MAIN RESULTS: After the scald injury followed by Staphylococcus aureus challenge, GTP-CHI messenger RNA expression and biopterin concentrations were significantly elevated in various tissues such as liver, heart, kidneys, and lungs, as were the values of inducible nitric oxide synthase messenger RNA expression and nitric oxide formation (p <.01). Pretreatment with DAHP significantly reduced GTP-CHI/biopterin induction (p <.05-.01), and the up-regulation of inducible nitric oxide synthase/nitric oxide was also suppressed. Furthermore, DAHP administration inhibited the gene expression of tumor necrosis factor-alpha. Two hours after septic challenge, tumor necrosis factor-alpha messenger RNA expression in liver, kidneys, and lungs in the DAHP-treated group was 35.7%, 37.3%, and 33.0% of that in the postburn septic group, respectively. Additionally, in animals without DAHP treatment, the 6-hr mortality rate was 55.6% (20 of 36), whereas it was only 25.0% in DAHP-treated animals (4 of 16, p =.08). CONCLUSIONS: Early treatment with DAHP might be a potential strategy to prevent the development of postburn Staphylococcal sepsis, which appears to be associated with down-regulation of biopterin and nitric oxide formation by DAHP.     

JAK/STAT通路介导脓毒症大鼠肝组织高迁移率族蛋白B1 mRNA表达的研究

目的：探讨Janusk激酶／信号转导和转录激活子（JAK／STAT）通路对盲肠结扎穿孔术（CLP）所致脓毒症大鼠肝组织高迁移率族蛋白B1（HMGB1）mRNA表达和急性肝损害的影响。方法：采用CLP模型，大鼠随机分为正常对照组、CLP脓毒症组、JAK2激酶抑制剂AG490和STAT抑制剂雷帕霉素（RPM）处理组。采用逆转录多聚酶链式反应测定肝HMGB1 mRNA，全自动生化分析仪测定肝功能指标。结果：与正常对照组相比，CLP后6－48h HMGB1 mRNA表达显著升高（P＜0．01）；血清天冬氨酸转氨酶（AST）在6－48h增高明显（P＜0．05），丙氨酸转氨酶（ALT）、AST在24h升高非常显著（P＜0．01）。与CLP组相比，AG490预处理组24h HMGB1 mRNA和ALT水平显著下降（P均＜0．01），24h和48h AST亦明显降低（P均＜0．01）；同样，RPM干预后HMGB1 mRNA表达在6h 和24h显著抑制（P＜0．05和P＜0．01），ALT、AST在24h和48h均不同程度下降（P＜0．01和P＜0．05）。结论：抑制JAK／STAT通路活化可明显下调肝组织HMGB1 mRNA表达，并有助于减轻CLP所致急性肝损伤。     

脓毒症大鼠肺高迁移率族蛋白B1的表达及意义

目的 观察创伤弧菌脓毒症大鼠肺组织HMGB1表达和肺损伤的动态变化,并探讨HMGB1在创伤弧菌脓毒症肺损伤中作用.方法 温州医学院生命科学院实验室,清洁级SD大鼠60只,随机分为正常组(A组,n=10)和创伤弧菌脓毒症组(B组,n=50),采用大鼠左下肢皮下注射创伤弧菌悬液(浓度为6×108cfu/mL,剂量为0.1 mL/100 g)制作大鼠创伤弧菌脓毒症模型),B组于染菌后1、6、12、24、48 h后活杀(各时间点n=10),采用逆转录聚合酶链式反应(RT-PCR)和蛋白免疫印迹(Western blot)分别检测大鼠肺组织HMGB1基因与蛋白的表达,检测肺含水分数和光镜观察肺组织病理变化,数据采用单因素方差分析,并用LSD法进行组间两两比较,P＜0.05为差异有统计学意义.结果 B组染菌后12 h(1.161±0.358,P=0.013)、24 h(1.679±0.235,P=0.000)及48 h(1.258±0.274,P=0.004)大鼠肺组织HMGB1 mRNA表达量较A组(0.652±0.177)明显增高(P＜0.05),并于24 h达到高峰;与A组(0.594±0.190)比较,B组HMGB1蛋白表达量于感染后6 h(1.408±0.567,P=0.026)(P＜0.05)逐渐增加,24 h达到高峰(2.415±1.064,P=0.000);与A组(0.699±0.054)比较,B组大鼠肺含水分数于感染后6 h(0.759±0.030,P=0.001)、12 h(0.767±0.023,P=0.000)、24 h(0.771±0.043,P=0.000)和48 h(0.789±0.137,P=0.000)明显增大(P＜0.05),呈逐渐递增趋势;感染后12 h,大鼠肺内血管明显充血,间质水肿并伴炎性浸润,且逐渐加重,到48 h肺泡腔塌陷明显,肺泡间隔分界不清.结论 大鼠创伤弧菌脓毒症可导致肺脏损伤,HMGB1的表达增加可能是创伤弧菌脓毒症大鼠肺组织损伤的机制之一.

Abstract：

Objective To observe the dynamic changes of high mobility group protein B1 ( HMGB1 )expression in the lung of rats with Vibrio vulnificus sepsis so as to unravel the role of HMGB1 in lung injury.Methods Sixty rats of clean grade were randomly divided into normal control group ( A group, n = 10) and Vibrio vulnificus sepsis group (B group, n =50). Sepsis model was made in rats with subcutaneous injection of Vibrio vulnificus with concentration of 6 × 108 cfu/ml in dose of 0. 1 ml/100 g into left lower limb.The rats of group B were sacrificed 1 h, 6 h, 12 h, 24 h and 48 h after infection for taking lung tissues to detect the water content of lung and to observe the histopathological changes in lung under light microscope.The expression of HMGB1 mRNA and the level of HMGB1 protein in the lungs were detected by RT-PCR and Western blot, respectively. Data were analysed with ANOVA and LSD method for comparison between groups, and P ＜0.05 was considered statistically significant. Results Compared with the group A (0.652±0. 177), the expressions of HMGB1 mRNA in lung of rats of group B were significantly higher in 12 hours (1. 161 ±0.358, P=0.013), 24 hours (1.679 ±0.235, P =0.000) and 48 hours (1.258 ±0.274, P=0.004) and reached the peak in 24 h. Compared with group A (0.594 ±0. 190), the level of HMGB1 protein in rats of group B 6 h after infection ( 1. 408 ± 0. 567, P = 0. 026) was significantly increased (P＜0.05), and it reached peak in 24 h (2.415 ± 1.064, P =0.000) after infection. Compared with group A (0.699 ± 0.054), the lung water contents in rats of group B were significantly increased in 6 h (0.759±0.030, P=0.001), in 12 h (0.767 ±0.023, P =0.000), in 24 h (0.771 ±0.043, P=0.000) and in 48 h (0.789 ±0.137, P=0.000) after infection. Compared with group A, the pathological changes in the lung of rats in group B showed clearly marked pulmonary vascular congestion, interstitial edema and inflammatory cell infiltration, and those changes became more and more serious until alveolar sacs entirely collapsed and the boundaries of the alveolar septa could not be clearly identified in 48 h. Conclusions Vibrio vulnificus sepsis leads to the lung injury of infected rats, and the increase in the expression of HMGB1 mRNA in lung might be one of the mechanisms of lung injury in rats with Vibrio vulnificus sepsis.     

Recombinant bactericidal/permeability-increasing protein inhibits endotoxin-induced high-mobility group box 1 protein gene expression in sepsis

We investigated in vivo the effect of recombinant bactericidal/permeability-increasing protein (rBPI21) on high-mobility group box 1 protein (HMGB1) expression in sepsis and its potential mechanism. Using a sepsis model induced by cecal ligation and puncture (CLP), rats were randomly divided into four groups as follows: normal control group, sham-operated group, CLP group, and BPI treatment group. Animals were killed at designated time points, and blood and tissue samples from liver, lungs, kidneys, and small intestine were harvested to determine related variables. In addition, we observed the effect of treatment with rBPI21 on survival rate in septic rats. The results showed that endotoxin content and expression levels of HMGB1 and LPS binding protein/CD14 mRNA in various organs were significantly increased at 12 and 24 h after CLP, which can be attenuated by treatment with rBPI21 (P<0.05-0.01). Meanwhile, treatment with rBPI21 in septic rats can markedly reduce serum alanine aminotransferase, creatinine levels, and pulmonary myeloperoxidase activity at 12 and 24 h after CLP, increase diamine oxidase activity at both time points (P<0.05-0.01), and improve the 1- to 10-day survival rates in animals subjected to CLP (P=0.012). These findings suggest that treatment with rBPI21 can significantly reduce endotoxin contents and expression levels of HMGB1 and LPS binding protein/CD14 mRNA in various organs in sepsis induced by CLP, and can protect against multiple organ damage resulting from sepsis. The effect of rBPI21 inhibiting HMGB1 gene expression in sepsis might be associated with endotoxin-dependent mechanisms.     

髓系细胞触发受体-1对重症急性胰腺炎大鼠肠屏障功能的影响

目的 探讨髓系细胞触发受体-1（triggering receptor-1 on myeloid cells,TREM-1）的表达与重症急性胰腺炎（severe acute pancreatitis,SAP）肠屏障功能障碍的关系。方法 雄性Wistar大鼠64只,随机（随机数字法）分为假手术组(SO)和SAP组,每组32只,采用逆行胰胆管注射5％牛磺胆酸钠制备SAP大鼠模型,分别于造模后2,6,12,24h时点取血和回肠组织。改良分光光度法检测血浆D-乳酸、二胺氧化酶（diamine oxidase,DAO）和内毒素浓度。逆转录-聚合酶链反应( RT-PCR)方法检测回肠组织TREM-1、白细胞介素-1β(interleukin-1β,IL-1β)和肿瘤坏死因子-α（tumor necrosis factor-α,TNF-α）mRNA的表达水平。对数据进行单因素方差分析和spearman相关性分析,P＜0.05表示差异具有统计学意义。结果SAP组各时点血浆D-乳酸、DAO和内毒素的水平均高于假手术组(P ＜0.01,P＜0.05);SAP组各时点回肠组织TREM-1,IL-1β和TNF-α mRNA的表达水平较假手术组显著增高(P ＜0.01,P＜0.05),TREM-1 mRNA表达水平与IL-1β及TNF-α mRNA表达水平均呈正相关(r=0.956,P =0.044;r =0.986,P=0.015),IL-1β mRNA表达水平与TNF-α mRNA表达水平无明显相关性(P=0.133)。结论 SAP时,大鼠肠组织内TREM-1表达上调,促进炎症介质释放和肠黏膜损伤加重,TREM-1在SAP肠屏障功能障碍的发生发展中起重要作用。     

Low-dose terlipressin improves systemic and splanchnic hemodynamics in fluid-challenged endotoxic rats

OBJECTIVE: Vasopressin has been used to treat arterial hypotension associated with hyperdynamic vasoplegic states, but detrimental effects on splanchnic circulation have been reported. We tested the effects of a low-dose vasopressin analogue, terlipressin (6 microg/kg), on systemic and splanchnic hemodynamics in fluid-challenged endotoxic rats (lipopolysaccharide, 30 mg/kg in 1 hr). DESIGN: Prospective, randomized, controlled experimental study with repeated measures. SETTING: Investigational animal laboratory. SUBJECTS: A total of 77 rats were divided into five groups: group C, control (17 rats); group E, LPS (18 rats); group EF, LPS plus fluid challenge (18 rats); group EFT, LPS plus fluid challenge plus terlipressin (18 rats); and group ET, LPS plus terlipressin (seven rats). INTERVENTIONS: Rats were anesthetized, mechanically ventilated, and instrumented to measure heart rate, mean arterial pressure, and abdominal aortic and mesenteric vein indexed blood flows; ileal microcirculation was assessed by laser Doppler. After LPS infusion, rats experienced an endotoxic shock and were resuscitated after the allocation group. The fluid challenge was targeted to maintain mean arterial pressure of >90 mm Hg and aortic blood flow at baseline values. MEASUREMENTS AND MAIN RESULTS: Terlipressin significantly (p <.05) increased mean arterial pressure without decreasing indexed aortic blood flow and heart rate in the fluid-challenged endotoxic rats (EFT) compared with EF rats and had detrimental effects in hypodynamic endotoxic rats (ET). Fluid challenge significantly (p <.05) increased mesenteric vein blood flow in both the EF and EFT groups, and terlipressin had no detrimental effect on mesenteric blood flow. Terlipressin significantly (p <.05) increased ileal microcirculation in fluid-challenged endotoxic rats (EF and EFT) but not in hypodynamic endotoxic rats (E and ET). CONCLUSION: Low-dose terlipressin in fluid-challenged endotoxic rats improved systemic and splanchnic hemodynamics and improved the ileal microcirculation.     

“717复方”制剂对内毒素损伤大鼠肝细胞线粒体的保护作用

目的：观察“７１７复方”制剂对内毒素休克大鼠肝细胞线粒体的保护作用。方法：采用静脉注射内毒素制备大鼠内毒素休克模型，观察“７１７复方”对休克大鼠肝细胞线粒体功能的影响。结果：模型组大鼠肝细胞线粒体呼吸控制率（ＲＣＲ）明显下降，与正常对照组比较（４．５８±０．３１比５．７３±０．３５）有显著性差异（Ｐ＜０．０５）；电镜下，内毒素休克模型线粒体明显肿胀，嵴减少、模糊不清。“７１７复方”制剂组大鼠肝细胞线粒体ＲＣＲ（５．６８±０．４１）则接近正常，与模型组比较有极显著性差异（Ｐ＜０．０１）；其电镜下超微结构病变也较轻。体外实验测定肝细胞线粒体膜通透性（膜吸光度下降百分比），“７１７复方”制剂组明显低于内毒素组（Ｐ＜０．０１）。结论：“７１７复方”制剂具有拮抗内毒素损伤线粒体的作用     

大黄抗内毒素性休克大鼠炎性介质作用的实验研究

目的：研究大黄对内毒素性休克大鼠炎性介质作用的机制。方法：选用大鼠内毒素性休克模型。随机分为６组：单纯手术组、内毒素组、大黄预防用药组（１５０ｍｇ／ｋｇ组和７５０ｍｇ／ｋｇ组）和大黄治疗组（１５０ｍｇ／ｋｇ组和７５０ｍｇ／ｋｇ组）。检测磷脂酶Ａ２（ＰＬＡ２）和血小板活化因子（ＰＡＦ）的活性。结果：内毒素注射前６组大鼠平均动脉压（ＭＡＰ）无显著性差异；注射内毒素后４小时ＭＡＰ明显降低；大黄预防用药组和大黄治疗组ＭＡＰ则与注射内毒素前及单纯手术组比较均无明显变化，并均显著高于内毒素组注射内毒素４小时后。注射内毒素后４小时，血清和小肠组织中ＰＬＡ２活性及ＰＡＦ含量均明显增高；与内毒素组注射内毒素后４小时比较，大黄预防组和治疗组则血清和小肠组织中ＰＬＡ２活性和ＰＡＦ含量显著降低。结论：大黄对内毒素性休克所致炎症反应有明显的预防和治疗作用