Elucidation of pathophysiology of ischemia reperfusion injury jn isolated perfused rat lungs

离体灌注大鼠肺缺血再灌注损伤病理生理学的阐明

• 批准号: 11470317
• 负责人: IMAI Takasuke
• 金额: $ 8万
• 依托单位: Tokyo Medical & Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470317/
• 关键词: ischemia-reperfusion; isolated rat lung; cyotokine; unilateral ischemia; inflammatory cytokines; RT-PCR; Inteferon-γ; Interferon-γ; 肺遊離環流標本; 虚血・再環流障害; 低酸素症; IL-1 beta; IL-6; 虚血・再潅流障害; 肺循環; 遊離潅流肺標本; mRNA

Isolated rat lung model perfused with Krebs-Henseleit solution added with 4% albumin was developed, in which right and left pulmonary arteries were isolated to enable preferential perfusion to each one of lungs with the same ventilation to the both lung maitained. Our hypothesis is that the genes of inflammatory cytokines would be induced in ischemic lung to cause the ischemia-reperfusion injury. After the left lung were exposed to ischemia for 60-100 min followed by reperfusion for 0-20 min, a few strips weighing about 100 mg were obtained separately in the perfused and ischemic lungs. Total RNA was extracted from these lung specimens by the method described by Chomczynski and Sacchi and DNAs of TNF-α, IL-1β, IL-10 and Interferon-γ were obtained by RT-PCR using MPCR primer, optimal temperatures and cycles. The obtained DNAs were electrophoresed in the agarose gel added with ethidium bromide. The genes of TNF-α, IL-1β and IL-6 in the control right lung lobe were detected more than in the left ischemic lung. Contrarily, the gene of IL-10 in the ischemic lung was detected more than in the control right lobe. Interfeorn-γ did not show significent difference between the both lobes. The reason why the inflammatory cyotokines appeared greater in the control right lung lobe than in the ischemic left lobe was supposed to the amount of endotoxin contained in the standard bovine serum albumin which were preferentially perfused through right lung during ischemia of left lung. Although the inflammatory cytokines were induced in the right and left lungs, left lung only showed ischemia reperfusion injury which suggested that interactions of any mediators induced by inflammatory cytokines and oxygen radicals would attack the cell membrane and cause the pulmonary edema.

开发了用添加了4％白蛋白的Krebs-Henseleit溶液灌注的分离的大鼠肺模型，其中隔离了左右肺动脉，以使每个肺都具有相同通风的每个肺对两个肺的灌注。我们的假设是，炎性细胞因子的基因将在缺血性肺中诱导，以引起缺血 - 再灌注损伤。将左肺暴露于缺血60-100分钟，然后再灌注0-20分钟后，在灌注和缺血性肺中分别获得了一些重约100 mg的条。通过使用MPCR引物，最佳温度和循环获得TNF-α，IL-1β，IL-10和Intferon-γ的Chomczynski和Sacchi和Sacchi和DNA所描述的方法从这些肺样本中提取总RNA。将所获得的DNA电泳在加入溴化乙锭的琼脂糖凝胶中。对照右肺叶中TNF-α，IL-1β和IL-6的基因比左侧缺血性肺更多。相反，与对照右叶中的缺血性肺中IL-10基因的检测到更多。 Itherfeorn-γ在两个裂片之间均未显示出显着差异。右肺叶中炎症性细性因子的表现比缺血性左叶更大的原因预期是标准牛血清白蛋白中包含的内毒素的量，在左肺缺血期间，通过右肺更喜欢通过右肺。尽管炎症性细胞因子是在左右肺部诱导的，但左肺仅显示缺血再灌注损伤，这表明炎性细胞因子和氧自由基引起的任何介质的相互作用会攻击细胞膜并引起肺部水肿。