Disruption of gut barrier function and cytokine responses after severe surgical stress-mechanism and treatment

严重手术应激机制和治疗后肠道屏障功能和细胞因子反应的破坏

• 批准号: 06671187
• 负责人: FUKUSHIMA Ryoji
• 金额: $ 1.34万
• 依托单位: Teikyo University School of Medicine (1995)The University of Tokyo (1994)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06671187/
• 关键词: Bacterial Translocation; Cytokines; Myeloperoxidase; Organ Injury; Growth Hormone (GH); Insulin-like Growth Factor I (IGF-I); 臓器障害; 顆粒球; bacterial translocatian; UGF-I; GH; IL-6

Experimenr IEffects of growth hormone (GH) and insulin-like growth factor I (IGF-I) administration on burn induced gut derived sepsis was evaluated. BALB/c mice (n=50) were treated subcutaneously with 4.8mg/kg/day of GH,24mg/kg/day of IGF-I or placebo twice a day for 4 days. Then they were gavaged with 10^<10>E.coli and subjected to 20% full sickness flame burn. All mice received allogeneic blood transfusion 5 days before burn to induce mild immunosuppression. Thirty mice were observed for survival and twenty mice were sacrificed at 20 hours post burn. GH and IGF-I administration reduced the incidence of translocation to MLN and other organs 20 hours post burn and significantly improved survival. We conclude that GH and IGF-I may be useful during critical illness.Experiment IIThe previous investigation revealed that bacterial translocation is highly associated with mortality after burn injury. To further clarify the mechanism of bacterial translocation related organ dysfunction and sub … More sequent death, we investigated the relation between the degree bacterial translocation and neutrophil accumulation in the liver.Blood transfused Balb/c mice were treated with oral 200mg/kg/day enisoprost (PGEl analog) or saline for three days and then they were gavaged with 10^<10 14>C E.coli and 20% full sickness flame burn was inflicted. According to our previous investigation, these mice have a mortality rate of approximately 80% and enisoprost improved the mortality to up to 30%. Animals were sacrificed 24 hour post burn and mesenteric lymph node (MLN), Liver, and spleen were harvested. Bacterial translocation were determined by both radionuclide count (dpm) and viable colony count in the MLN and Liver. Leukocyte accumulation was evaluated by the measurement of myeloperoxidase (MPO) in the liver. Consistent to previous work, enisoprost significantly reduced the translocation. MPO in the liver was significantly greater in the control group compared to enisoprost group. There was a significant correlation between MPO and the degree of bacterial translocation (p<0.05). It can concluded that bacterial translocation enhanced neutrophil accumulation in the liver which may be the cause of organ injury after burn injury.Experiment IIIUsing the same model as experiment II,splenic macrophages were separated and cultured for 24 hours with and without 10mcg/ml of LPS.TNF,IL-1 IL-6 and PGE2 in the cell culture supernatants were measured. LPS stimulated macrophage production of IL-1, IL-6 and PGE2 were significantly greater in enisoprost treated animals. It is likely that prior lack of in vivo maximal stimulation of macrophages in the enisoprost treated animals can produce greater amounts of cytokines when further stimulated with LPS in vitro. Less

实验一观察了生长激素（GH）和胰岛素样生长因子I（IGF-I）对烧伤后肠源性脓毒症的治疗作用。BALB/c小鼠（n=50）用4.8mg/kg/天的GH、24 mg/kg/天的IGF-I或安慰剂皮下处理，每天两次，持续4天。然后，用10 μ L大肠杆菌管饲它们<10>，并使它们经受20%的完全疾病火焰烧伤。所有小鼠在烧伤前5天接受同种异体输血以诱导轻度免疫抑制。观察30只小鼠的存活情况，并在烧伤后20小时处死20只小鼠。GH和IGF-I的管理减少易位到MLN和其他器官的发生率烧伤后20小时，显着提高生存。我们的结论是，GH和IGF-I可能是有用的，在危重病。实验IIThe以前的调查显示，细菌易位是高度相关的烧伤后死亡率。为进一步阐明细菌移位相关器官功能障碍的机制， ...更多信息 给Balb/c小鼠灌胃200 mg/kg/d的前列腺素（PGE 1）或生理盐水，连续3天，然后用10 μ <10 14>C大肠杆菌灌胃，造成20%的全身火焰烧伤。根据我们之前的研究，这些小鼠的死亡率约为80%，而依异前列素将死亡率提高至30%。烧伤后24小时处死动物，收获肠系膜淋巴结（MLN）、肝脏和脾脏。通过放射性核素计数（dpm）和MLN和肝脏中的活菌落计数来确定细菌移位。通过测量肝脏中的髓过氧化物酶（MPO）来评价白细胞蓄积。与先前的工作一致，依异前列素显著减少了易位。对照组肝脏中的MPO显著高于依异前列素组。MPO与细菌移位程度呈显著相关（p&lt;0.05）。实验三采用与实验二相同的模型，分离脾巨噬细胞，在加入和不加入10 mcg/ml LPS的情况下培养24小时，测定细胞培养上清中TNF、IL-1、IL-6和PGE_2的含量。LPS刺激的巨噬细胞产生的IL-1，IL-6和PGE 2显着更大的治疗的动物。可能的是，先前在艾尼前列素处理的动物中缺乏巨噬细胞的体内最大刺激，当在体外用LPS进一步刺激时，可以产生更大量的细胞因子。少