Cao Ying-Ya, Wang Zhong-Han, Xu Qian-Cheng, Chen Qun, Wang Zhen, Lu Wei-Hua
Department of Intensive Care Unit, The First Affiliated Hospital of Wannan Medical College, Wuhu 241001, China.
Korean J Physiol Pharmacol. 2021 Jul 1;25(4):375-383. doi: 10.4196/kjpp.2021.25.4.375.
The intestinal barrier function disrupted in sepsis, while little is known about the variation in different phases of sepsis. In this study, mouse models of sepsis were established by caecal ligation and puncture (CLP). The H&E staining of sections and serum diamine oxidase concentration were evaluated at different timepoint after CLP. TUNEL assay and EdU staining were performed to evaluate the apoptosis and proliferation of intestinal epithelium. Relative protein expression was assessed by Western blotting and serum concentrations of pro-inflammatory cytokines was measured by ELISA. The disruption of intestinal barrier worsened in the first 24 h after the onset of sepsis and gradually recovered over the next 24 h. The percentage of apoptotic cell increased in the first 24 h and dropped at 48 h, accompanied with the proliferative rate of intestinal epithelium inhibited in the first 6 h and regained in the later period. Furthermore, the activity of nuclear factor kappa B (NF-κB) presented similar trend with the intestinal barrier function, shared positive correction with apoptosis of intestinal epithelium. These findings reveal the conversion process of intestinal barrier function in sepsis and this process is closely correlated with the activity of NF-κB signaling.
脓毒症时肠道屏障功能受损，而关于脓毒症不同阶段的变化情况却知之甚少。在本研究中，通过盲肠结扎和穿刺（CLP）建立脓毒症小鼠模型。在CLP后的不同时间点评估切片的苏木精-伊红（H&E）染色和血清二胺氧化酶浓度。进行TUNEL检测和EdU染色以评估肠上皮细胞的凋亡和增殖情况。通过蛋白质免疫印迹法评估相关蛋白表达，并通过酶联免疫吸附测定（ELISA）测量促炎细胞因子的血清浓度。脓毒症发作后的最初24小时内肠道屏障破坏加剧，在接下来的24小时内逐渐恢复。凋亡细胞百分比在最初24小时增加，在48小时下降，同时肠上皮细胞的增殖率在最初6小时受到抑制，在后期恢复。此外，核因子κB（NF-κB）的活性与肠道屏障功能呈现相似趋势，与肠上皮细胞凋亡呈正相关。这些发现揭示了脓毒症中肠道屏障功能的转变过程，且该过程与NF-κB信号通路的活性密切相关。
