栀子苷通过调控单核细胞表型改善小鼠盲肠结扎穿孔技术脓毒症模型预后

目的:探讨不同剂量栀子苷治疗脓毒症的疗效和主要生物学机制。方法:雄性BALB/c小鼠通过盲肠结扎穿孔技术(cecal ligation and puncture, CLP)复制脓毒症模型。在生存实验中,动物被随机分为以下各组,每组20只：于CLP术后0 h、24 h经小鼠尾静脉分别注射栀子苷20 mg/kg、40 mg/kg或生理盐水(对照组);于CLP术后24 h经小鼠尾静脉注射栀子苷40 mg/kg或生理盐水(对照组)。观察不同组别的生存预后;并流式检测单核细胞CD16、MHC-Ⅱ、TLR2、TLR4表达水平;ELISA检测血清TNF-α、IL-1β、IL-6、IL-10浓度;Western Blot测定PPARγ浓度。结果:40 mg/kg栀子苷CLP后0 h和24 h静脉给药能显著改善脓毒症小鼠模型生存预后,小剂量(20 mg/kg)栀子苷和延迟给药(24 h)无显著获益。与对照组相比,有效剂量栀子苷能不同程度地全面抑制脓毒症小鼠血清细胞因子TNF-α、IL-1β、IL-6、IL-10浓度,差异有统计学意义(P<0.05);能降低脓毒症小鼠24 h单核细胞CD16表达,差...     

缺血／再灌注联合盲肠结扎穿孔术大鼠肠系膜血管白细胞与内皮细胞的相互作用

失血性休克导致的缺血／再灌注（HS／R）损伤常诱发脓毒症和微循环紊乱，甚至多器官功能障碍综合征（MODS）。巴西学者最近进行了一项研究，对大鼠实施HS／R联合盲肠结扎穿孔术（CLP），随后行盲肠切除加腹腔灌洗（REL），观察肠系膜血管中白细胞与内皮细胞的相互作用。实验中将81只Wistar大鼠（200～250g）随机分为3组：①对照组：无HS／R损伤；②HS／R损伤Ⅰ组：HS加25％失血量的血液进行再灌注；③HS／R损伤Ⅱ组：HS加25％失血量的血液以及3倍失血量的乳酸林格液（LRS）再灌注。3组动物24h后全部行CLP，再过24h行REL。     

脓毒症大鼠肠道肠三叶因子mRNA表达的变化

目的：探讨脓毒症大鼠肠道肠三叶因子（trefoil factor family,TFF3）mRNA表达的变化。方法：32只SD大鼠随机分为对照组（n=8）和脓毒症组（n=24）。采用盲肠结扎穿孔术（cecal ligation and puncture,CLP）制作大鼠脓毒症模型。于模型建立后3h,6h及12h（n=8）取回肠黏膜,以RT-PCR法检测TFF3 mRNA的表达。结果：脓毒症模型建立3h始TFF3 mRNA表达即显著下降（P〈0.01）,随着时间延长表达进一步下降。结论：脓毒症大鼠肠道TFF3 mRNA表达明显下降,可能是脓毒症肠屏障功能障碍的机制之一并延缓肠道黏膜修复。     

阻滞交感神经改善脓毒症大鼠肠道微循环

因胃肠道局部缺血、缺氧、黏膜屏障作用受损导致的肠道内细菌／毒素移位与脓毒症发生发展密切相关，在生理条件下阻滞交感神经可提高肠道灌注压。瑞士学者研究了硬膜外麻醉阻滞交感神经对脓毒症大鼠肠黏膜微循环障碍的影响。研究者采用大鼠盲肠结扎穿孔术制备脓毒症模型，然后经胸椎硬膜外导管连续24h给予质量分数为0.125％的布比卡因（n=10）和生理盐水（n=9），     

717复方制剂对小白鼠盲肠结扎穿孔术后所致感染性休克的影响

选择了小白鼠盲肠结扎穿孔所致感染性休克的实验模型，探索中医药开闭法（代表药７１７复方制剂）抗感染性休克的作用机制。结果：７１７复方制剂组小白鼠盲肠结扎穿孔术后的存活时间为２８．７５±５．０６小时，明显长于对照组（饮用水组）１２．５５±０．６３小时（Ｐ＜０．０５）；心脏组织匀浆中丙二醛（ＭＤＡ）为２３９．６２±１２．４１ｎｍｏｌ／ｍｇｐｒｏ，低于对照组２９７．８８±１７．０１ｎｍｏｌ／ｍｇｐｒｏ（Ｐ＜０．０５）；肝脏组织匀浆中ＭＤＡ为２７２．４１±５．９２ｎｍｏｌ／ｍｇｐｒｏ，明显低于对照组４２８．７７±１５．１４ｎｍｏｌ／ｍｇｐｒｏ，（Ｐ＜０．０１）。说明７１７复方制剂是通过抗氧化而起保护作用的。     

山莨菪碱对脓毒症大鼠肺组织生物喋呤的影响

目的：探讨山莨菪碱对脓毒症大鼠肺组织生物喋呤产生的影响及其意义。方法：采用大鼠盲肠结扎穿孔（ＣＬＰ）致脓毒症模型,动物随机分为正常对照组、脓毒症组和山莨菪碱治疗组。分别于ＣＬＰ后２、８、１６小时处死动物,检测肺组织生物喋呤及其合成限速酶－－三磷酸鸟苷环水解酶Ⅰ（ＧＴＰ－ＣＨⅠ）ｍＲＮＡ表达和肿瘤坏死因子－α（ＴＮＦ－α）ｍＲＮＡ表达的改变。结果：脓毒症动物肺组织生物喋呤含量显著升高,ＧＴＰ－ＣＨＩ     

免气腹腹腔镜胃十二指肠溃疡穿孔修补术13例分析

消化性溃疡穿孔是普外科常见的急诊手术之一，我院自2005年5月至2007年11月利用免气腹腹腔镜行胃十二指肠穿孔修补术，共13例，现报道如下：     

硫化氢在脓毒症大鼠肺损伤中的作用

目的 探讨硫化氢(H2S)在脓毒症大鼠肺损伤中的作用.方法 雄性SD大鼠60只,随机分为4组:对照组(Ⅰ组)15只,脓毒症组(Ⅱ组)15只,脓毒症+PAG(H2S代谢酶CSE抑制剂)组(Ⅲ组)15只,脓毒症+NaHS(H2S供体)预处理组(Ⅳ组)15只.采用盲肠结扎穿孔法制作脓毒症大鼠模型,观察各组动物的行为学特征,测定肺组织H2S浓度、肺组织CSE mRNA的表达、肺组织湿干重比(W/D)、肺组织MPO水平、肺组织TNF-α和IL-1β的表达,观察肺组织形态学改变.结果 CLP模型组动物出现呼吸加快,与Ⅰ组比较,Ⅱ组动物肺组织H2S、W/D、TNF-α、IL-1β水平明显升高(P＜0.01),MPO活性明显增加(P＜0.01),肺组织CSE mRNA的表达明显升高(P＜0.01);与Ⅱ组比较,Ⅲ组动物呼吸频率接近,肺组织H2S、W/D、TNF-α、IL-1β水平明显降低(P＜0.01),MPO活性明显减弱(P＜0.01),肺组织CSE mRNA的表达明显降低(P＜0.01);与Ⅱ组比较,Ⅳ组动物呼吸频率明显加快,少数出现呼吸衰竭,肺组织W/D、TNF-α、IL-1β水平明显升高(P＜0.01),MPO活性明显增强(P＜0.01),肺组织CSE mRNA的表达有所降低,但差异无统计学意义(P＞0.05);四组肺组织光镜下损伤由轻到重依次为:Ⅰ、Ⅲ、Ⅱ、Ⅳ.结论 给予内源性H2S抑制剂可以使脓毒症大鼠肺组织W/D、TNF-α、IL-1β、MPO水平明显降低,减轻肺组织炎症及水肿损伤,给予外源性H2S可以加重肺炎症损伤.     

鞭毛蛋白与盲肠结扎穿孔脓毒症致大鼠肺损伤的对比观察

目的 观察鞭毛蛋白与盲肠结扎穿孔脓毒症致大鼠肺损伤的差异。方法 鞭毛蛋白经颈静脉注射以复制大鼠急性肺损伤(ALI)模型。ELISA法检测大鼠外周血血清中TNF-α含量,并观察动脉血气分析、肺组织湿干比值(W/D)和肺病理变化程度。结果 用鞭毛蛋白成功建立了大鼠肺损伤模型。鞭毛蛋白致肺损伤大鼠的动脉氧分压(PaO2)、TNF-α、W/D和肺病理变化程度较脓毒症致肺损伤大鼠有显著性差别。结论 鞭毛蛋白可用于急性肺损伤模型的建立。     

脓毒症大鼠心肌细胞凋亡及糖皮质激素的作用

目的 观察脓毒症状态下大鼠心肌细胞凋亡的变化并探讨糖皮质激素对心肌细胞凋亡的影响及其机制,为临床治疗脓毒症心脏损害提供依据.方法 采用盲肠结扎穿孔术制作脓毒症模型.60只体质量为230 ～ 280 g的Wistar大鼠被随机(随机数字法)分为糖皮质激素实验组(n=30)和对照组(n=30).每组大鼠均行盲肠结扎穿孔术,每组术后4h开始给予舒普深200 mg/kg,2次/d.实验组在以上基础上给予地塞米松0.5 mg/kg.每组随机分为3个亚组(6、24、72 h组),于相应时间点取出各组大鼠心脏,分离左心室,制备单细胞悬液,进行流式细胞仪检测心肌细胞凋亡率.使用SPSS 11.5统计软件系统,所有数据以均数±标准差表示((x-)±s),两组间采取独立样本t检验,多组间数据比较采用单因素方差分析.同时左心室行免疫组化检测抗细胞凋亡蛋白B淋巴细胞瘤/白血病-2(B-cell lymphoma/leukemia-2,Bcl-2)的表达.结果 激素组大鼠各时间点心肌细胞凋亡率分别较对照组大鼠心肌细胞凋亡率明显减少(F=9.11,t=5.681,P＜0.01)(6 ht =11.416,P＜0.01;24 h=6.217,P＜0.01;72 h t=3.76,P＜0.01);6h对照组大鼠的心肌细胞凋亡率显著低于24 h及72 h对照组大鼠(F=13.254,sig =0.000,P＜0.01;sig=0.004,P＜0.01);24 h对照组大鼠的心肌细胞凋亡率显著高于72 h对照组大鼠(sig=0.039,P＜0.05);激素各组大鼠心肌细胞凋亡率差异无统计学意义(F=2.488,6/24 h sig=0.132,P＞0.05; 24/72 h sig=0.549,P＞0.05;6/72 h sig=0.053,P＞0.05).结论 脓毒症时存在心肌细胞凋亡,以24 h最明显,脓毒症时应用糖皮质激素能够减少各个阶段心肌细胞凋亡.     

Cecal ligation and puncture induced sepsis impairs host defense against Enterococcus faecium peritonitis

Purpose  Multiresistant and vancomycin resistant Enterococcus faecium (VRE) can cause serious infections in hospitalized patients with various co-morbid diseases. We investigated the course of VRE peritonitis after cecal ligation and puncture (CLP)-induced sepsis and compared this to sham operated mice. Methods  Mice were subjected to CLP or sham surgery. Forty-eight hours thereafter four groups were created by subjecting mice to peritoneal injection of either VRE or saline. Results  Mice infected with VRE after CLP were severely impaired in eliminating VRE from the peritoneal cavity and distant body sites. These mice failed to mount an early inflammatory response at the primary site of VRE infection. VRE superinfection did not influence CLP-induced organ damage or polymicrobial bacterial loads. Conclusions  Sublethal polymicrobial sepsis greatly facilitates infection and dissemination of VRE. VRE does not influence the course of CLP-induced sepsis.     

δ阿片受体激动剂D-丙2,D-亮5脑啡肽对脓毒症大鼠死亡率的影响

目的观察δ阿片受体激动剂D-丙2,D-亮5脑啡肽（[D-Ala2,D-Leu5]enkephalin,DADLE）对脓毒症大鼠存活时间和死亡率的影响。方法采用盲肠结扎穿孔（CLP）方法制备大鼠脓毒症模型。120只SD大鼠（雌雄不拘）,按随机数字表法分为假手术组（n=20）、模型对照组（n=20）,及4个DADLE治疗组。各DADLE组按给药剂量及方式分为制模前给药3mg/kg组（n=20）、制模前给药5mg/kg组（n=20）、制模后给药3mg/kg组（n=20）、制模后给药5mg/kg组（n=20）。观察各组大鼠CLP术后的存活时间,计算7d累积死亡率。结果与CLP模型未治疗组比较,各DADLE治疗组大鼠CLP术后的存活时间显著延长（P〈0.05）,各组7d死亡率分别为85%、60%、70%及75%,均较CLP组明显降低 但各DADLE治疗组之间无统计学差异（P〉0.05）。结论δ阿片受体激动剂DADLE能够明显延长脓毒症大鼠的存活时间,降低脓毒症大鼠的死亡率。     

Imipramine reverses the depressive symptoms in sepsis survivor rats

Objective To evaluate the antidepressant effect of imipramine on depressive symptoms observed in sepsis survivors rats. Design and setting Prospective, controlled experiment in an animal basic science laboratory. Subjects Male Wistar rats weighing 300–350 g. Interventions The rats underwent cecal ligation and perforation (CLP; sepsis group) with “basic support” (saline at 50 ml/kg immediately and 12 h after CLP plus ceftriaxone at 30 mg/kg and clindamycin at 25 mg/kg 6, 12, and 18 h after CLP) or sham-operated (control group). After 10 days of recovery rats received intraperitoneal injections of imipramine 10 mg/kg or saline and were subjected to the forced swimming test. Measurements and results The observed increase in the immobility time in the forced swimming test in animals subjected to CLP, as a parameter of depressive behavior, was reversed by imipramine. Conclusions The depressive symptoms evaluated by forced swimming test had been reversed after imipramine administration. Our data provide evidence that CLP-induced depressive symptoms are sensitive to antidepressants. This research was supported in part by UNESC (Brazil), FAPESC (Brazil), and CNPq (Brazil).     

脓毒症早期大鼠下丘脑-垂体-肾上腺轴超微结构变化与功能的关系

目的 观察脓毒症早期下丘脑-垂体-肾上腺(HPA)轴结构及功能的变化,并探讨其相互关系.方法 雄性SD大鼠30只,按随机数字表法均分为正常对照组、假手术组和脓毒症组.采用盲肠结扎穿孔术(CLP)制作脓毒症大鼠模型,术后6 h取血并处死动物,检测血浆促肾上腺皮质激素(ACTH)、皮质酮(CoRT)以及下丘脑促肾上腺皮质激素释放激素(CRH)水平;透射电镜下观察HPA轴的超微结构改变.结果 脓毒症组血浆ACTH、CORT和下丘脑CRH水平均明显高于正常对照组和假手术组[ACTH(pmol/L):5.78±0.36比1.94±0.31、2.51±0.10;CORT(nmol/L):88.48±4.47比22.02±1.62、34.20±2.51,CRH(μg/L):101.92±6.61比61.65±6.05、66.65±4.03,P＜0.05或P＜0.01].透射电镜下观察:脓毒症组大鼠下丘脑粗面内质网囊状扩张、脱颗粒,高尔基体肿胀,垂体ACTH细胞分泌颗粒增多,肾上腺组织脂滴减少.结论 脓毒症早期大鼠HPA轴处于过度激活状态,血浆ACTH、CORT和下丘脑CRH水平明显升高;HPA轴的超微结构明显改变,且与功能变化有密切联系.

Abstract：

Objective To observe the changes in ultrastructure and function of hypothalamicpituitary-adrenal axis (HPAA), and to approach the relationship between them in early stage of sepsis in rats. Methods Thirty male Sprague-Dawley (SD) rats were randomly divided into normal control group,sham group, sepsis group. The sepsis model was reproduced by cecal ligation and puncture (CLP). The rats were sacrificed after collection of blood at 6 hours after CLP, and the levels of adrenocorticotropic hormone (ACTH) and corticosterone (CORT) in the plasma, and the corticotropin release hormone (CRH) in the tissue of hypothalamus were detected. The histopathological changes in HPAA were observed with transmission electron microscopy. Results The levels of ACTH and CORT in plasma, and the CRH in hypothalamus tissue of sepsis group were increased in the early stage of sepsis compared with the normal control group or sham group [ACTH (pmol/L): 5. 78 ± 0. 36 vs. 1.94 ±0.31, 2. 51 ± 0.10; CORT (nmol/L), 88.48±4.47 vs. 22.02±1.62, 34.20±2.51; CRH (μg/L): 101. 92±6. 61 vs. 61.65±6.05,66. 65±4. 03, P＜0. 05 or P＜0. 01]. The changes in ultrastructure of the hypothalamus, pituitary and adrenal were also found. In sepsis group, the ultrastructure of hypothalamus was as follows. Rough endoplasmic reticulum expansion and degranulation of rough endoplasmic reticulum, and swelling of Golgi complex were found. A large number of endocrine granules could be seen in ATCH cells in the pituitary with depletion of adrenal lipid droplets. Conclusion In septic rats, the HPAA was excessively activated, and ACTH and CORT in plasma, and CRH in hypothalamus were significantly increased in early stage of sepsis.The changes in ultrastructure of HPAA were obvious, and the change in function was closely related to the ultrastructural changes.     

Esmolol reduces apoptosis and inflammation in early sepsis rats with abdominal infection

Background

Esmolol is a highly selective beta 1 receptor blocker with various effects such as slowing heart rate, lowering blood pressure and reducing myocardial oxygen consumption. However, few studies have reported the use of beta blockers in sepsis with multiple organ dysfunctions. This study aimed to investigate the effects of esmolol on reducing apoptosis and inflammation in early sepsis rats with abdominal infection.

乌司他丁对脓毒症大鼠肠道防御素5 mRNA表达的影响

目的 探讨乌司他丁(ulinastatin,LTI)对脓毒症大鼠肠道潘氏细胞防御素5(rat defemin-5,RD-5)mRNA表达的影响.方法 实验在中山大学医学院药理实验室完成.60只SD大鼠随机分为对照组、脓毒症组、预处理组及治疗组(n=15).后三组采用盲肠结扎穿孔术(cecal ligation andpuncture,CLP)制作大鼠脓毒症模型.预处理组在CLP前2h经尾静脉注射UTI 25 000 U/kg,治疗组在CLP后2 h经尾静脉注射UTI 50 000 U/kg.于模犁建立后12 h取回肠黏膜,观察其病理改变并以RT-PCR法检测RD-5 mRNA的表达.数据以SPSS 13.0统计软件处理,采用方差分析及LSD-t检验进行数据分析.结果 RD-5 mRNA在脓毒症组显著下降(P<0.05),预处理组及治疗组较脓毒症组有明显升高(P<0.05),预处理组较治疗组明显升高(P<0.05).结论 脓毒症大鼠RD-5mRNA表达明显下降,乌司他丁可显著上调其表达,保护肠黏膜,预防用药较治疗给药可能更有意义.     

脓毒症大鼠肠黏膜免疫屏障功能的变化

目的 研究脓毒症对大鼠肠黏膜免疫屏障功能的影响.方法 60只SD大鼠随机(随机数字法)分为对照组(n=15)和脓毒症组(n=45),采用盲肠结扎穿孔术(CLP)建立脓毒症模型.模型建立后3 h、6 h和12 h留取回肠黏膜和全血标本.分别进行肠黏膜形态学观察、肠防御素5(RD-5)及肠三叶因子3(TFF_3)Mrna表达水平检测、肠黏膜淋巴细胞凋亡分析,以及外周血中肠源性细菌DNA定性检测.结果 CLP所致脓毒症导致大鼠回肠黏膜明显损害,主要表现为上皮脱落、固有层分离、毛细血管出血和溃疡形成;脓毒症组模型建立后3 h即出现RD-5和TFF_3 Mrna表达显著性减少(与正常组比较,P<0.05),且6 h和12 h组进行下降(与3 h组比较,P<0.05),肠黏膜淋巴细胞凋亡数亦显著增加(P<0.05);同时,脓毒症组全血肠源性细菌DNA扩增全部阳性.结论 脓毒症时大鼠肠黏膜免疫屏障功能显著减退,且随脓毒症的发展而进行性恶化.     

Changes of the immunological barrier of intestinal mucosa in rats with sepsis

BACKGROUND:

Sepsis has become the greatest threat to in-patients, with a mortality of over 25%. The dysfunction of gut barrier, especially the immunological barrier, plays an important role in the development of sepsis. This dysfunction occurs after surgery, but the magnitude of change does not differentiate patients with sepsis from those without sepsis. Increased intestinal permeability before surgery is of no value in predicating sepsis. The present study aimed to observe the changes of intestinal mucosal immunologic barrier in rat models of sepsis induced by cecal ligation and puncture.

METHODS:

Sixty Sprague-Dawley rats were randomly divided into a sepsis group (n=45) and a control group (n=15). The rats in the sepsis group were subjected to cecal ligation and puncture (CLP), whereas the rats in the control group underwent a sham operation. The ileac mucosa and segments were harvested 3, 6 and 12 hours after CLP, and blood samples were collected. Pathological changes, protein levels of defensin-5 (RD-5) and trefoil factor-3 (TFF₃) mRNA, and lymphocytes apoptosis in the intestinal mucosa were determined. In an additional experiment, the gut-origin bacterial DNA in blood was detected.

RESULTS:

The intestinal mucosa showed marked injury with loss of ileal villi, desquamation of epithelium, detachment of lamina propria, hemorrhage and ulceration in the sepsis group. The expression of TFF₃ mRNA and level of RD-5 protein were decreased and the apoptosis of mucosal lymphocyte increased (P<0.05) in the sepsis group compared with the control group. Significant differences were observed in RD-5 and TFF₃ mRNA 3 hours after CLP and they were progressively increased 6 and 12 hours after CLP in the sepsis group compared with the control group (P<0.05, RD-5 F=11.76, TFF₃ F=16.86 and apoptosis F=122.52). In addition, the gut-origin bacterial DNA detected in plasma was positive in the sepsis group.

CONCLUSION:

The immunological function of the intestinal mucosa was impaired in septic rats and further deteriorated in the course of sepsis.KEY WORDS: Sepsis, Mucosal immunology, Defensin-5, Trefoil factor family 3, Cecal ligation and puncture     

Repair of damaged intestinal mucosa in a mouse model of sepsis

BACKGROUND:

The intestine is not only the main target attacked by sepsis but also the vital organ which mediated sepsis. The recovery of the damaged intestinal barrier structure and function is related to the occurrence and outcome of multiple organ dysfunction syndrome (MODS). How to protect and reduce the damage of the intestinal mucosa and how to promote the reconstruction of the intestinal mucosa have been the important topics in sepsis for many years. This study aimed to investigate the influential factors of intestinal mucosal reconstruction after intestinal epithelial injury in vivo in a mouse model of sepsis.

METHODS:

Mice were subjected to cecal ligation and puncture (CLP) for induction of sepsis to assess intestinal mucosal damage, epithelial cell apoptosis, and transformed number of goblet cells, and to detect the concentration of TNF-α, IL-1 and TGF-β1 and TFF3 (trefoil factor 3) expression in the small intestinal mucosa. All above were performed by HE staining, western blot, ELISA and immunohistochemistry respectively. The experimental animals were divided into a sepsis group and a sham-operation group. The animals with sepsis were separately killed at 6 (7 animals), 24 (7 animals) and 48 hours (7 animals) after CLP.

RESULTS:

Injured intestinal mucosa was observed in the 3 groups under a light microscope, in which damage scores in the 24-hour and 48-hour groups were higher than in the 6-hour group and no difference was found between the two groups. Moreover, less of goblet cells or other epithelial cells adjacent to the injured surface migrated into the wound to cover the denuded area. The number of goblet cells was substantially decreased in the three CLP groups compared with the sham-operation group. Protein levels of IL-1 and TNF-α were significantly increased by 3–4 fold at all time points when compared with the sham-operation group, and cleaved caspase-3 by 4 fold. Although TFF3 expression was modestly increased for 6 hours after the onset of CLP, it appeared to decline at 24 hours and 48 hours as shown by Western blot. A similar tendency was observed upon TGF-β1, i.e. the protein level was not elevated at 24 hours and 48 hours, but increased modestly at 6 hours.

CONCLUSIONS:

Sepsis from CLP shows less restitution on the surface of injured intestinal mucosa. There is evidence that both constant inflammatory reaction and epithelial cell apoptosis may affect mucosal reestablishment of the intestine at the onset of sepsis. Mucosa after severe sepsis showed the state of high inflammation, and declined goblet cell function and mucosal reconstruction, which affected the repair of damaged intestinal barrier. Constant inflammatory reaction, and declined goblet cell function and mucosal reconstruction ability may affect the reestablishment of intestinal mucosa at the onset of sepsis.KEY WORDS: Sepsis, Cecal ligation and puncture, Intestinal mucosa, Restitution, Goblet cells, Intestinal trefoil factor 3, Transforming growth factor β1, Cysteine-containing aspartate-specific proteases     

影响脓毒症受损肠黏膜修复的机制

目的 探讨影响脓毒症肠黏膜损害后修复的因素。方法 采用肓肠结扎穿孔(CLP)所致脓毒症模型,分别以CLP后6,24,48 h不同时间段观测肠黏膜损伤程度和修复过程,前者包括形态学观察及细胞凋亡的测定,后者包括肠黏膜修复的杯状细胞变化、黏膜肠三叶因子3(TFF3)、转化生长因子β1(TGF-β1)以及TNF-α、IL-1含量。结果 形态学观察显示肠黏膜呈持续损害状态,6h的损害积分明显小于24h,48 h组(P＜0.05),后两组之间差异无统计学意义(P＞0.05);磷酸化caspase-3蛋白在3组均高于sham组4倍以上;黏膜IL-1,TNF-α含量明显高于sham组3～4倍,其中24h及48 h组明显高于6h组。肠黏膜的修复过程不明显,损伤黏膜未见到明显的杯状细胞积聚;TFF3在6h组轻度增高,24h及48 h组表达下降;杯状细胞数量在CLP的3个组明显减少;TGF-β1在6h组增高,其他两组均接近于sham组。结论 严重脓毒症肠黏膜持续的高炎症状态、杯状细胞功能以及黏膜重建能力下降,影响了受损肠屏障的修复。