创伤后肠道菌群移位与微生态防治研究（一）：烧伤后内源性感染症的动物模…

为了深入地研究肠道菌群的生物屏障作用,我们使用Ｗｉｓｔａｒ大鼠制备烧伤动物模型,这亘地分析了烧伤大鼠肠道粘膜和内容物中５类菌群,且对血液和内脏进行了细菌培养,结果显示烧伤大鼠４８ｈ菌群变化明显,烧伤大鼠４８ｈ血培养阳性率普５０％,细菌移位率达４１％,烧伤２４ｈ血浆内毒素升高,二胺氧化酶活性和结肠ＩｇＡ水平下降,发生了内源性感染症。     

双歧杆菌活菌制剂对烧伤鼠肠源性感染的预防作用

本文以３０％烫伤同时口服抗生素,喂饲双歧杆菌制剂的大鼠为模型,将Ｗｉｓｔａｒ大鼠分为烧伤治疗组,烧伤对照组及正常对照组。并于伤后２４、４８、７２、９６小时分别观察盲肠内容物菌群,粘蛋白,分泌型免疫球蛋白Ａ（ＳＩｇＡ）;肠道细菌移位率;腔静脉内毒素含量。结果发现烧伤后肠道菌群有不同程度的变化,念珠菌迅速过度生长、厌氧菌下降、肠内需厌氧菌比例严重失调,治疗组中粘蛋白含量,肠道细菌移位率均明显小于烧伤对照组;而ＳＩｇＡ含量则高于对照组,菌群改变较对照组轻。故认为双歧杆菌制剂有助于预防烧伤肠源性感染的发生。     

鱼油对断奶大鼠脏器指数及肠道菌群影响的研究

目的探讨鱼油对断奶大鼠脏器指数及肠道菌群的影响。方法选取（21±3）d日龄断奶SD大鼠96只,雌雄各半,随机分为2组,实验组饲喂添加0．5％鱼油的饲料,对照组饲喂正常饲料。分别于第7、14、21、28天处死各组大鼠12只,分别测定大鼠肝脏指数、胸腺指数及脾脏指数;应用梯度稀释法和培养法测定大鼠4种肠道正常菌群,即肠杆菌、双歧杆菌、乳酸杆菌、葡萄球菌。结果实验组第7天脾脏指数与对照组相比差异有显著性（P〈0．05）,第28天肝脏指数和胸腺指数与对照组相比差异有显著性（P〈0．05）;实验组第7、14和21天肠道内肠杆菌数量与对照组相比差异有显著性（P〈0．05）,第7天大鼠肠道内葡萄球菌数量与对照组相比差异有显著性（P〈0．05）,第14天大鼠肠道内葡萄球菌数量与对照组相比差异有非常显著性（P〈0．01）,实验组大鼠肠道内乳酸杆菌和双歧杆菌数量与对照组相比虽有所增高,但差异无显著性（P〉0．05）。结论鱼油可以明显提高断奶大鼠的脏器指数,并且可通过增加断奶大鼠肠道内双歧杆菌和乳酸杆菌数量,降低肠道内肠杆菌和葡萄球菌数量,进而调节断奶大鼠的肠内菌群,改善肠道内环境。     

基于16S rRNA基因和细菌基因组间重复序列的DNA指纹图谱技术对肝硬化大鼠肝移植后肠道菌群多样性的研究

目的应用PCR-DGGE和rep-PCR技术对肝硬化大鼠肝移植后肠道菌群多样性进行研究,探讨肠道菌群多样性的变化,并比较这两种方法在菌群分析中的作用。方法首先建立CCL4诱导肝硬化大鼠的肝移植模型,收取对照组、肝硬化成模时、肝移植后7 d和肝移植后30 d的大鼠粪便,提取细菌基因组DNA,采用PCR-DGGE和rep-PCR[BOX-PCR,ERIC-PCR,ERIC2-PCR,(GTG)5-PCR,REP-PCR]进行DNA指纹图谱分析。结果PCR-DGGE可明确将正常大鼠、肝硬化大鼠、肝移植大鼠分为3个簇,并显示出肝硬化、肝移植大鼠肠道菌群多样性明显增多。rep-PCR技术也可将各组分开,其中ERIC-PCR、ERIC2-PCR及REP-PCR三者扩增条带各组差异有显著性,鉴别效果更好。结论应用基于16S rRNA基因和细菌基因组间重复序列的指纹图谱技术对肝硬化大鼠肝移植后肠道微生态的研究具有重要价值,可对临床肝硬化、肝移植患者肠道微生态制剂的使用起到指导作用。     

肠道菌群在应激诱发大鼠高血压中的作用

观察应激性高血压大鼠肠道菌群的变化,探索肠道菌群在应激性高血压发生中的作用。应激性高血压模型的制备:大鼠每天2次接受足底电刺激加噪声的应激,每次2 h,中间间隔4 h,共14 d。血压的测量:大鼠股动脉插管,Powerlab系统记录血压。伪无菌鼠的制备:在大鼠的饮用水中加入氨苄西林(1 g/L)、万古霉素(500 mg/L)、新霉素(1 g/L)和甲硝唑(1 g/L)4种抗生素,共持续4周。采用Illumina MiSeq测序技术,测定大鼠粪便中微生物16S rRNA V3-V4区序列,并对群落结构和多样性进行比较分析。慢性应激导致大鼠血压升高,心率加快,血清去甲肾上腺素浓度增加;α多样性分析显示应激大鼠肠道菌群丰富度和多样性下降,β多样性分析显示应激组大鼠肠道菌群物种组成变少,差异性变大;伪无菌鼠的肠道菌群物种单一,仅有少量的变形菌门。给予伪无菌鼠相同的慢性应激,不能引起大鼠血压升高、心率增快,血清去甲肾上腺素浓度未升高。应激引起大鼠肠道菌群紊乱,应激性高血压的发生可能依赖于肠道菌群的状态。     

益生菌对烧伤大鼠肠道膜菌群和sIgA的影响

目的探索益生菌对严重烧伤大鼠早期肠道菌群的调理作用,为防治肠源性感染寻找新途径.方法选用健康Wistar大鼠100只,体重180～220 g,雌雄各半,随机分为:益生菌治疗组(BR),烧伤对照组(BC)和正常对照组(NC),建立30% Ⅲ°烫伤肠源性感染的动物模型,按时分批活杀取材,检测盲肠膜菌群和肠黏膜sIgA的含量.结果 BC组盲肠中双歧杆菌数量与NC组比较,明显减少(P＜0.01),而BR组与NC组比较,差异无显著性(P＞0.05);BC组肠道中酵母菌和大肠埃希菌与NC比较明显增加,BR组差异无显著性;肠粘液sIgA水平与上述指标有类似变化.结论 30%Ⅲ°烫伤大鼠肠道内容物中,双歧杆菌明显下降,大肠埃希菌、酵母菌迅速过生长,导致肠道微生态失衡,应用BR治疗后,促进肠黏膜机械屏障功能的恢复,调理肠道微生态平衡,对肠黏膜起占位性保护作用,提高了肠道局部和全身益生菌免疫的功能.     

SD大鼠断奶后肠道菌群及肠道形态的变化

目的通过对断奶日龄不同的SD大鼠肠道菌群和肠道形态学指标变化的观察研究,为临床上研究肠道相关性疾病提供参考数据。方法选取断奶日龄为21 d的SD大鼠60只,雌雄各半,分别在断奶后第0、7、14、21、28天测定4种肠道正常菌群及肠道黏膜形态变化。结果SD大鼠断奶后不同时间点,不同肠段内大肠杆菌、双歧杆菌、乳酸杆菌和葡萄球菌呈现不同的变化规律,且断奶后第7天肠道细菌移位率明显提高,同时断奶后不同时间点,大鼠不同肠段黏膜厚度、肌层厚度、绒毛长度和宽度都呈现上升趋势。结论断奶后第7天和14天,SD大鼠肠道菌群和肠道形态学变化明显,且易发生肠道细菌移位。     

大鼠肥胖抑郁模型建立及肠道菌群多样性的性别差异研究

目的 肥胖抑郁大鼠模型建立并对比研究肠道菌群多样性的变化。方法 采用高脂饲料喂养和CUMS经典造抑郁方法建立肥胖抑郁模型。将SD大鼠随机分为正常组(C组)、单纯肥胖组(F组)、单纯抑郁组(Y组)和肥胖抑郁组(FY组),整个造模持续5个月，通过糖水偏好实验、旷场实验和强迫游泳实验来评估大鼠抑郁样行为，检测模型鼠血清雌激素水平和血脂4项，并采集各组大鼠粪便样品，通过16S rDNA高通量测序法和生物信息分析软件分析测序结果，比较各组大鼠肠道菌群多样性的变化。结果 高脂饮食和CUMS方法成功制作肥胖抑郁鼠模型，肥胖和肥胖抑郁雌性大鼠雌激素水平明显升高，而雄性各组间雌激素水平无显著性差异。雌雄鼠的FY组肠道菌群α多样性具有显著差别，拟杆菌门和厚壁菌门B/F比值下降，且雄鼠肥胖抑郁B/F值只有雌鼠的一半左右。在属水平，研究结果表明雌雄鼠FY组和F组肠道菌群中Blautia等8个菌群变化一致，但是雌性与雄性在拟杆菌属等各有少量菌群的变化不一致。雌雄肥胖抑郁模型中，肠道菌群革兰氏阳性菌和阴性菌的组成出现差异，G⁺的构成，雌性和雄性均由由厚壁菌门和放线菌门构成，但构成比例不一样。...     

16S rDNA测序研究高脂饮食诱导的高脂血症大鼠肠道菌群变化

目的 通过高脂饮食诱导大鼠高脂血症,采用16S rDNA测序检测高脂血症大鼠肠道菌群变化情况。方法 SD大鼠20只(清洁级),按体质量随机分为模型组和对照组,对照组大鼠给予维持饲料,模型组大鼠给予高脂饲料。1周后检测血清总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)和高密度脂蛋白胆固醇(HDL-C)水平。采集大鼠粪便,采用16S rDNA测序法对大鼠肠道菌群进行分析,考察高脂血症大鼠肠道菌群变化情况。结果 高脂血症大鼠肠道菌群生物多样性(Alpha多样性和Beta多样性)发生显著变化,肠型与对照组差异明显,在门、科、属等多个水平差异均具有统计学意义。其中,厚壁菌门(Firmicutes)和念珠菌门(Candidatus saccharibacteria)数量显著下降,拟杆菌门(Bacteroidetes)、疣微菌门(Verrucomicrobia)、变形菌门(Proteobacteria)和放线菌门(Actinobacteria)数量显著升高。结论 高脂饮食诱导的高脂血症可引起大鼠肠道菌群发生显著变化。     

新生鼠坏死性小肠结肠炎肠道菌群变化及意义

目的探讨新生鼠坏死性小肠结肠炎(NEC)时肠道菌群变化,旨在阐明肠道菌群在NEC发病中的作用,为探讨新生儿NEC的发病机制及寻求有效的防治措施提供理论依据。方法SD新生大鼠出生48 h开始给予鼠乳代用品人工喂养,100%氮气缺氧90 s,4℃冷刺激10 min,每天2次,连续3 d,建立新生SD大鼠NEC模型。40只新生SD大鼠随机分成NEC模型组(A组)和正常对照组(B组)。每组动物各20只。在最后一次缺氧、冷刺激后24 h空腹断头处死大鼠,留取回盲部近端肠管组织进行肠组织损伤评分,组织学评分≥2确定为NEC;实验前后留取两组新生鼠粪便,按照张秀荣方法进行肠道菌群检测。实验前后比较采用配对t检验,组间比较采用方差分析,α=0.05为显著性检验标准。结果模型组新生鼠相继出现腹泻、腹胀、萎靡、活动减少,生长减慢,对照组新生大鼠进食及排便均正常,无腹胀及胃潴留,活动度良好,皮下脂肪丰满。模型组新生大鼠NEC的发生率为100%(20/20),对照组无1例发生NEC。实验组和对照组肠损伤病理评分(x±s)分别为:3.25±0.85、0.45±0.51,t=12.622,P<0.01。模型组和对照组新生鼠实验前肠道细菌总数,杆菌、球菌总数,G 杆菌、G 球菌,G-杆菌、G-球菌数差异均无显著性,肠道菌群中杆菌和球菌的比例都在正常范围中。实验结束时,正常对照组新生大鼠肠道菌群总数明显增多,其中以G 杆菌增加为主;G-杆菌属及G 球菌属菌数占肠道菌群的比率在实验前后差异无显著性;模型组新生大鼠肠道群总数亦明显增多,而且明显高于正常对照组。实验结束时肠道菌群数量,其中主要是G 球菌显著增加,而G 杆菌却较对照组明显减少,与实验前相比,明显下降,差异有显著性;模型组实验结束时肠道菌群中杆菌和球菌比值减少、倒置。结论NEC发病前正常肠道菌群已发生质和量的变化,肠道菌群紊乱在NEC发病机制中起关键作用;及时纠正肠道菌群紊乱可能会降低新生大鼠发生NEC危险性。     

Redistribution of blood flow after thermal injury and hemorrhagic shock

Diminished mucosal mass and a diminished rate of DNA synthesis by the intestinal mucosa have been identified in the rat after thermal injury. Because these changes may be associated with ischemia, the distribution of intestinal blood flow was studied after a thermal injury and compared with the blood flow distribution after hemorrhagic shock. For the thermal injury, anesthetized animals received a standardized 20% body surface area, full-thickness injury and were given intraperitoneal saline resuscitation. By the use of 46Sc- or 141Ce-labeled microspheres, no changes in intestinal and hepatic blood flow occurred after thermal injury. In contrast, a marked redistribution of blood flow was identified after hemorrhagic shock in which a decrease in arterial blood flow was identified to the stomach and to the small and large intestine. Although clinical shock was not present, the cardiac output decreased to a comparable degree in the hemorrhagic shock and the thermal injury. These studies indicate that although physiological changes in intestinal mucosa can be demonstrated after burn injury, these changes are not due to decreases in mesenteric arterial blood flow.     

Alcohol ingestion before burn injury decreases splanchnic blood flow and oxygen delivery

Recent studies from our laboratory have shown that alcohol and burn injury impair intestinal barrier and immune functions. Although multiple factors can contribute to impaired intestinal barrier function, such an alteration could result from a decrease in intestinal blood flow (BF) and oxygen delivery (DO2). Therefore, in this study, we tested the hypothesis that alcohol ingestion before burn injury reduces splanchnic blood flow and oxygen delivery. Rats (250 g) were gavaged with alcohol to achieve a blood ethanol level in the range of 100 mg/dl before burn or sham injury (25% total body surface area). Day 1 after injury, animals were anesthetized with methoxyflurane. Blood pressure, cardiac output (CO), +/-dP/dt, organ BF (in ml.min(-1).100 g(-1)), and DO2 (in mg.ml(-1).100 g(-1)) were determined. CO and organ BF were determined using a radioactive microsphere technique. Our results indicate that blood pressure, CO, and +dP/dt were decreased in rats receiving a combined insult of alcohol and burn injury compared with rats receiving either burn injury or alcohol alone. This is accompanied by a decrease in BF and DO2 to the liver and intestine. No significant change in BF to the coronary arteries (heart), brain, lung, skin, and muscles was observed after alcohol and burn injury. In conclusion, the results presented here suggest that alcohol ingestion before burn injury reduces splanchnic BF and DO2. Such decreases in BF and DO2 may cause hypoxic insult to the intestine and liver. Although a hypoxic insult to the liver would result in a release of proinflammatory mediators, a similar insult to the intestine will likely perturb both intestinal immune cell and barrier functions, as observed in our previous study.     

Bombesin improves burn-induced intestinal injury in the rat

This study was designed to determine the effect of exogenous bombesin (10 microg/kg/day, subcutaneously, three times a day) on intestinal hypomotility and neutrophil infiltration in the early and late phases of burn injury (partial-thickness, second-degree burn of the skin). In acute (2 h after burn injury) or chronic (3 days after) burn groups, intestinal transit was delayed, which was reversed by bombesin treatment. In the acute burn group, but not in the chronic group, increased MPO activity was also reduced by bombesin treatment. The results demonstrate that bombesin ameliorates the intestinal inflammation due to burn injury, involving a neutrophil-dependent mechanism.     

Myosin light chain kinase mediates intestinal barrier disruption following burn injury

Background

Severe burn injury results in the loss of intestinal barrier function, however, the underlying mechanism remains unclear. Myosin light chain (MLC) phosphorylation mediated by MLC kinase (MLCK) is critical to the pathophysiological regulation of intestinal barrier function. We hypothesized that the MLCK-dependent MLC phosphorylation mediates the regulation of intestinal barrier function following burn injury, and that MLCK inhibition attenuates the burn-induced intestinal barrier disfunction.

Methodology/Principal Findings

Male balb/c mice were assigned randomly to either sham burn (control) or 30% total body surface area (TBSA) full thickness burn without or with intraperitoneal injection of ML-9 (2 mg/kg), an MLCK inhibitor. In vivo intestinal permeability to fluorescein isothiocyanate (FITC)-dextran was measured. Intestinal mucosa injury was assessed histologically. Tight junction proteins ZO-1, occludin and claudin-1 was analyzed by immunofluorescent assay. Expression of MLCK and phosphorylated MLC in ileal mucosa was assessed by Western blot. Intestinal permeability was increased significantly after burn injury, which was accompanied by mucosa injury, tight junction protein alterations, and increase of both MLCK and MLC phosphorylation. Treatment with ML-9 attenuated the burn-caused increase of intestinal permeability, mucosa injury, tight junction protein alterations, and decreased MLC phosphorylation, but not MLCK expression.

Conclusions/Significance

The MLCK-dependent MLC phosphorylation mediates intestinal epithelial barrier dysfunction after severe burn injury. It is suggested that MLCK-dependent MLC phosphorylation may be a critical target for the therapeutic treatment of intestinal epithelial barrier disruption after severe burn injury.     

Heme oxygenase-1 protects against neutrophil-mediated intestinal damage by down-regulation of neutrophil p47phox and p67phox activity and O2- production in a two-hit model of alcohol intoxication and burn injury

Heme oxygenase-1 (HO-1) has been demonstrated to protect against tissue injury. Furthermore, HO-1 is also shown to be antioxidant. Our recent findings indicate that acute alcohol (EtOH) intoxication exacerbates postburn intestinal and lung tissue damage, and this was found to be neutrophil dependent. Because neutrophil-mediated tissue injury involves the release of superoxide anions (O(2)(-)), the present study examined the role of HO-1 in neutrophil O(2)(-) production following EtOH and burn injury. Furthermore, we investigated whether HO-1 antioxidant properties are mediated via modulation of p47(phox) and/or p67(phox) proteins. Male rats (approximately 250 g) were gavaged with EtOH to achieve a blood EtOH level of approximately 100 mg/dL before burn or sham injury (approximately 12.5% total body surface area). Some rats were treated with HO-1 activator cobalt protoporphyrin IX chloride (Copp; 25 mg/kg body weight) at the time of injury. On day 1 after injury, we found that EtOH combined with burn injury significantly increased neutrophil O(2)(-) production and p47(phox) and p67(phox) activation and decreased caspase-3 activity and apoptosis. This was accompanied with a decrease in neutrophil HO-1 levels. The treatment of animals with HO-1 activator Copp normalized neutrophil HO-1, O(2)(-), p47(phox), and p67(phox) following EtOH and burn injury. The expression of caspase-3, however, was further decreased in Copp-treated sham and EtOH plus burn groups. Moreover, Copp treatment also prevented the increase in intestinal edema and permeability following EtOH and burn injury. Altogether, these findings provide a new insight into the mechanism by which HO-1 regulates neutrophil O(2)(-) production and protect the intestine from damage following EtOH and burn injury.     

Gut-associated lymphoid T cell suppression enhances bacterial translocation in alcohol and burn injury

The mechanism of alcohol-mediated increased infection in burn patients remains unknown. With the use of a rat model of acute alcohol and burn injury, the present study ascertained whether acute alcohol exposure before thermal injury enhances gut bacterial translocation. On day 2 postinjury, we found a severalfold increase in gut bacterial translocation in rats receiving both alcohol and burn injury compared with the animals receiving either injury alone. Whereas there were no demonstrable changes in intestinal morphology in any group of animals, a significant increase in intestinal permeability was observed in ethanol- and burn-injured rats compared with the rats receiving either injury alone. We further examined the role of intestinal immune defense by determining the gut-associated lymphoid (Peyer's patches and mesenteric lymph nodes) T cell effector responses 2 days after alcohol and burn injury. Although there was a decrease in the proliferation and interferon-gamma by gut lymphoid T cells after burn injury alone; the suppression was maximum in the group of rats receiving both alcohol and burn injuries. Furthermore, the depletion of CD3(+) cells in healthy rats resulted in bacterial accumulation in mesenteric lymph nodes; such CD3(+) cell depletion in alcohol- and burn-injured rats furthered the spread of bacteria to spleen and circulation. In conclusion, our data suggest that the increased intestinal permeability and a suppression of intestinal immune defense in rats receiving alcohol and burn injury may cause an increase in bacterial translocation and their spread to extraintestinal sites.     

Alteration in intestine tight junction protein phosphorylation and apoptosis is associated with increase in IL-18 levels following alcohol intoxication and burn injury

Intestinal mucosal barrier is the first line of defense against bacteria and their products originating from the intestinal lumen. We have shown a role for IL-18 in impaired gut barrier function following acute alcohol (EtOH) intoxication combined with burn injury. To further delineate the mechanism, this study examined whether IL-18 alters intestine tight junction proteins or induces mucosal apoptosis under these conditions. To accomplish this, rats were gavaged with EtOH (3.2 g/kg) prior to ~ 12.5% total body surface area burn or sham injury. One day after injury, EtOH combined with burn injury resulted in a significant decrease in total occludin protein and its phosphorylation in small intestine compared to either EtOH or burn injury alone. There was no change in claudin-1 protein content but its phosphorylation on tyrosine was decreased following EtOH and burn injury. This was accompanied with an increase in mucosal apoptosis (p < 0.05). The treatment of rats with anti-IL-18 antibody at the time of burn injury prevented intestine apoptosis and normalized tight junction proteins following EtOH and burn injury. Altogether, these findings suggest that IL-18 modulates tight junction proteins and cause apoptosis leading to impaired intestinal mucosal integrity following EtOH intoxication combined with burn injury.     

Inhibition of long myosin light-chain kinase activation alleviates intestinal damage after binge ethanol exposure and burn injury

Laboratory evidence suggests that intestinal permeability is elevated following either binge ethanol exposure or burn injury alone, and this barrier dysfunction is further perturbed when these insults are combined. We and others have previously reported a rise in both systemic and local proinflammatory cytokine production in mice after the combined insult. Knowing that long myosin light-chain kinase (MLCK) is important for epithelial barrier maintenance and can be activated by proinflammatory cytokines, we examined whether inhibition of MLCK alleviated detrimental intestinal responses seen after ethanol exposure and burn injury. To accomplish this, mice were given vehicle or a single binge ethanol exposure followed by a sham or dorsal scald burn injury. Following injury, one group of mice received membrane permeant inhibitor of MLCK (PIK). At 6 and 24 h postinjury, bacterial translocation and intestinal levels of proinflammatory cytokines were measured, and changes in tight junction protein localization and total intestinal morphology were analyzed. Elevated morphological damage, ileal IL-1β and IL-6 levels, and bacterial translocation were seen in mice exposed to ethanol and burn injury relative to either insult alone. This increase was not seen in mice receiving PIK after injury. Ethanol-exposed and burn-injured mice had reduced zonula occludens protein-1 and occludin localization to the tight junction relative to sham-injured mice. However, the observed changes in junctional complexes were not seen in our PIK-treated mice following the combined insult. These data suggest that MLCK activity may promote morphological and inflammatory responses in the ileum following ethanol exposure and burn injury.     

卡巴胆碱改善犬50%TBSA烧伤胃内补液时氧动力学指标

目的:研究卡巴胆碱(CAR)对50%TBSA烧伤休克期胃内补液时氧动力学指标的影响。方法:成年雄性Beagle犬12只,先期无菌手术行颈动、静脉置管和肠造口术,24h后用凝固汽油燃烧法造成50%体表面积Ⅲ度烧伤。随机分为2组(n=6):胃内补液组和胃内补液+CAR组。伤后第一个24h从胃内分别输注葡萄糖-电解质溶液(GES)和含CAR的GES液(20μg/kgCAR溶于GES);伤后24h起实施静脉延迟补液,补液量和速率均根据Park-land公式确定。测定动物非麻醉状态下的平均动脉压(MAP)、肠粘膜血流量(IMBF)和血乳酸(LAC)含量,通过血气分析测定动、静脉氧分压和血氧含量、计算氧供量(DO2)、氧耗量(VO2)和氧摄取(Oext),并统计动物72h死亡数。结果:两组犬MAP和IMBF伤后均显著降低,LAC显著升高;伤后72hMAP回升至0h水平,但IMBF和LAC仍低于或高于0h水平。伤后2h胃内补液+CAR组MAP显著高于胃内补液组(P0.01),但之后两组MAP水平无统计学差别。胃内补液/CAR组IMBF伤后高于胃内补液组,伤后8h起LAC也显著低于胃内补液组(P0.05或P0.01)。伤后两组犬DO、VO2和Oext水平较伤前均显著降低(P0.01),伤后72hDO2恢复至0h水平,但VO2和Oext仍显著低于0h。胃内补液/CAR组DO2、VO2和Oext水平伤后8h起始终高于胃内补液组(P0.05或P0.01)。伤后72h胃内补液组死亡数为3/6,胃内补液/CAR为2/6。结论:50%TBSA烧伤胃内补液时加入卡巴胆碱能显著改善氧动力学指标,降低高乳酸血症,提高口服液体复苏的疗效。     

Corticosterone suppresses mesenteric lymph node T cells by inhibiting p38/ERK pathway and promotes bacterial translocation after alcohol and burn injury

Previous studies showed that alcohol (EtOH) intoxication before burn injury suppresses mesenteric lymph node (MLN) T cell functions and increases gut bacterial translocation. In this study, we examined whether corticosterone (Cort) plays any role in suppressing MLN T cell function and bacterial accumulation after EtOH intoxication and burn injury. Rats were gavaged with EtOH to achieve a blood EtOH level of approximately 100 mg/dl before receiving 25% total body surface area burn or sham injury. A group of rats was treated with the Cort synthesis inhibitor metyrapone (25 mg/kg) at the time of injury and on day 1 after injury. Two days after injury, a significant increase in blood Cort levels and suppression of MLN T cell proliferation and IL-2 production was observed in rats receiving combined insult of EtOH intoxication and burn injury compared with rats receiving EtOH intoxication or burn injury alone. There was no change in T cell apoptosis after combined insult of EtOH and burn injury. Furthermore, T cell suppression was accompanied by a significant decrease in p38 and ERK1/2 activation (phosphorylation). There was no difference in JNK activation after EtOH and burn injury. Treatment of rats with metyrapone prevented the suppression of MLN T cell proliferation, IL-2 production, and p38 and ERK1/2 phosphorylation. Restoration of T cell function in metyrapone-treated animals was also associated with the decrease in bacterial accumulation in MLN. These findings suggest that EtOH intoxication before burn injury augments Cort release, which suppresses MLN T cell function by inhibiting p38 and ERK1/2 activation and promotes bacterial accumulation in MLN after EtOH and burn injury.