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投稿时间：2024-09-03 网络发布日期：2024-11-20
投稿时间：2024-09-03 网络发布日期：2024-11-20
中文摘要: 目的 探讨乌司他丁（UTI）对脓毒症诱导的急性肺损伤（ALI）大鼠氧化应激水平以及肠道菌群的影响，为脓毒症ALI的诊治提供理论支持。 方法 将18只雄性Wistar大鼠按随机数字表法分为假手术组（Sham组）、盲肠结扎穿孔术（CLP）致脓毒症组（CLP组）和脓毒症+UTI干预组（CLP+UTI组，CLP术后即刻腹腔注射UTI 100 kU/kg），每组6只。造模后24 h处死大鼠取肺组织，光镜下观察肺组织病理学改变；采用酶联免疫吸附试验（ELISA）检测肺组织中超氧化物歧化酶（SOD）活性和丙二醛（MDA）含量；蛋白质免疫印迹试验（western blotting）法测定大鼠肺组织核因子E2相关因子2（Nrf2）和血红素加氧酶-1（HO-1）的表达；采用实时荧光定量聚合酶链反应（RT-qPCR）法检测Nrf2、HO-1、肿瘤坏死因子α（TNF-α）和白细胞介素-6（IL-6）的mRNA表达；采用16S rRNA基因测序检测大鼠肠道菌群的变化。结果 与Sham组相比，CLP组的组织病理学结果显示肺泡有渗出、炎性细胞浸润、肺组织结构紊乱；肺组织炎症因子TNF-α和IL-6的mRNA表达均升高，SOD活性降低，MDA含量升高（P＜0.05）；肺组织Nrf2和HO-1的mRNA及蛋白表达降低（P＜0.05）。与CLP组相比，CLP+UTI组肺组织损伤明显缓解，肺组织炎症因子TNF-α和IL-6的mRNA表达显著降低（P＜0.05），SOD活性升高［SOD（U/mg prot）：11.83±0.99 vs 6.74±0.43，P＜0.05］，MDA含量降低［MDA（nmol/mg prot）: 0.16±0.04 vs 0.38±0.03，P＜0.05］；肺组织 Nrf2和HO-1的mRNA及蛋白表达升高。结论 乌司他丁对脓毒症诱导的急性肺损伤有明显保护作用，可能通过调节肠道菌群进而改善炎症损伤，并减少氧化应激损伤。
Abstract:Objective To investigate the effect of ulinastatin (UTI) on the level of oxidative stress as well as intestinal flora in rats with sepsis-induced acute lung injury (ALI), providing theoretical support for the diagnosis and treatment of sepsis-induced ALI. Methods Eighteen male Wistar rats were divided into three groups: sham operation group (Sham group), cecum ligation and puncture (CLP) induced sepsis group (CLP group), and sepsis+UTI intervention group (CLP+UTI group, intraperitoneal injection of 100 kU/kg UTI instantly after CLP), with 6 rats in each group. Lung tissues were obtained from rats executed 24 hours after modeling, and pathological changes were observed under the light microscope. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content were detected by enzyme linked immunosorbent assay (ELISA). The expression of nuclear factor erythroid 2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) was measured by western blotting. The mRNA expression of Nrf2, HO-1, tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) was detected by real-time quantitative polymerase chain reaction (RT-qPCR). The changes of intestinal flora in rats were detected by 16S rRNA gene sequencing. Results Compared with the Sham group, histopathologic analysis observed that the lungs of the CLP group exhibited alveolar effusion, inflammatory cell infiltration, and structural disorganization; the mRNA expression of TNF-α and IL-6 were significantly elevated in lung tissue, SOD activity decreased, the content of MDA increased; the mRNA and protein expressions of Nrf2 and HO-1 in lung tissue were decreased. Compared with the CLP group, the lung tissue damage in the CLP+UTI group was significantly relieved, the mRNA expression of TNF-α and IL-6 were significantly decreased in lung tissue (P＜0.05), SOD activity increased ［SOD (U/mg prot): 11.83±0.99 vs 6.74±0.43, P＜0.05］, the content of MDA decreased ［MDA (nmol/mg prot): 0.16±0.04 vs 0.38±0.03, P＜0.05］; the mRNA and protein expressions of Nrf2 and HO-1 in lung tissue were increased (P＜0.05). Conclusion UTI has a significant protective effect against sepsis-induced ALI, possibly by modulating the intestinal flora which in turn ameliorates inflammatory injury and reduces oxidative stress injury.
文章编号： 中图分类号：R631 文献标志码：A
基金项目:新疆维吾尔自治区自然科学基金项目（2022D01C477）；国家“十四五”重点专项（2021YFC2500800）；新疆维吾尔自治区科技支疆项目（2021E02064）
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引用文本：
张怡人,陈梦晓,李祥,等.乌司他丁对脓毒症诱导的急性肺损伤大鼠氧化应激和肠道菌群的影响[J].中国临床研究,2024,37(11):1674-1679.
张怡人,陈梦晓,李祥,等.乌司他丁对脓毒症诱导的急性肺损伤大鼠氧化应激和肠道菌群的影响[J].中国临床研究,2024,37(11):1674-1679.