TLR4/MyD88/NF-κB 在乌司他丁对脓毒症小鼠肠黏膜屏障保护作用中的作用。
The role of TLR4/MyD88/NF-κB in the protective effect of ulinastatin on the intestinal mucosal barrier in mice with sepsis.
机构信息
Department of Anesthesiology, Shaanxi Provincial Hospital, The Third Affiliated Hospital of Xi'an JiaoTong University, Xi'an, 710068, Shaanxi Province, People's Republic of China.
Xi'an Medical University, Xi'an 710068, Shaanxi Province, People's Republic of China.
出版信息
BMC Anesthesiol. 2023 Dec 15;23(1):414. doi: 10.1186/s12871-023-02374-9.
OBJECTIVE
To investigate the effect of the TLR4/MyD88/NF-κB (Toll-like receptor 4/myeloid differentiation factor/nuclear factor kappa B) signalling pathway on the protective effect of ulinastatin on the intestinal mucosal barrier in mice with sepsis.
METHODS
A mouse model of sepsis was established by classical caecal ligation and perforation. Forty-four SPF C57BL/6 mice were randomly divided into the following four groups with 11 mice in each group: the control group (Con group), ulinastatin group (Uti group), Uti + LPS (lipopolysaccharide, LPS) group (Uti + LPS group) and LPS group. Mice in the Con group and Uti group received saline or ulinastatin injected 2 h after modelling; Mice in the Uti + LPS group received LPS injected 0 h after modelling, other procedures were the same as in the Uti group; Mice in the LPS group received LPS only. At 48 h after surgery, the levels of TNF-α (tumour necrosis factor-α, TNF-α), IL-6 (interleukin-6, IL-6) and IL-1β (interleukin-1β, IL-1β) in vein, and the expression of TLR4, MyD88 and NF-κB mRNA in small intestinal mucosa tissues using ELISA and RT‒PCR.
RESULTS
The pathological specimens showed increased inflammatory injury in the Con and LPS groups, while these injuries and changes improved in the Uti group. The scores of intestinal mucosal injury at 48 h of Uti injection were significantly lower than those of the Con group (P < 0.001), while the scores of intestinal mucosal injury of Uti + LPS were significantly higher than those of the Uti group (P = 0.044). The expression of TNF-α, IL-6 and IL-1β in the Uti decreased significantly at 48 h after surgery than that in the Con group (P = 0.001, P = 0.014, P = 0.004), while the expression of TNF-α, IL-6 and IL-1β in the Uti + LPS group increased significantly after surgery than that in the Uti group (P = 0.026, P = 0.040, P = 0.039). The expression of TLR4, MyD88 and NF-κB mRNA in the Uti group decreased significantly compared with that in the Con group (P = 0.001, P = 0.021, P = 0.007), while the expression of TLR4, MyD88 and NF-κB mRNA in the Uti + LPS group was higher than that in the Uti group (P = 0.023, P = 0.040, P = 0.045).
CONCLUSION
These findings indicate that the protective effect of ulinastatin on the intestinal mucosal barrier against sepsis may be mediated through the TLR4/MyD88/NF-κB pathway.
目的
探讨 Toll 样受体 4/髓样分化因子 88/核因子 κB（Toll-like receptor 4/myeloid differentiation factor 88/nuclear factor kappa B，TLR4/MyD88/NF-κB）信号通路对乌司他丁对脓毒症小鼠肠道黏膜屏障保护作用的影响。
方法
采用经典盲肠结扎穿孔法建立脓毒症小鼠模型。将 44 只 SPF C57BL/6 小鼠随机分为以下 4 组，每组 11 只：对照组（Con 组）、乌司他丁组（Uti 组）、乌司他丁+脂多糖（lipopolysaccharide，LPS）组（Uti+LPS 组）和 LPS 组。Con 组和 Uti 组小鼠造模后 2 h 给予生理盐水或乌司他丁注射；Uti+LPS 组小鼠造模后 0 h 给予 LPS 注射，其他操作同 Uti 组；LPS 组仅给予 LPS。术后 48 h，采用 ELISA 和 RT-PCR 法检测静脉血中 TNF-α、IL-6 和 IL-1β 水平，以及小肠黏膜组织中 TLR4、MyD88 和 NF-κB mRNA 的表达。
结果
病理标本显示 Con 组和 LPS 组炎症损伤增加，而 Uti 组的损伤和变化得到改善。乌司他丁注射后 48 h 时，Uti 组的肠黏膜损伤评分明显低于 Con 组（P<0.001），而 Uti+LPS 组的肠黏膜损伤评分明显高于 Uti 组（P=0.044）。术后 48 h，Uti 组 TNF-α、IL-6 和 IL-1β 的表达明显低于 Con 组（P=0.001、P=0.014、P=0.004），而 Uti+LPS 组 TNF-α、IL-6 和 IL-1β 的表达明显高于 Uti 组（P=0.026、P=0.040、P=0.039）。Uti 组 TLR4、MyD88 和 NF-κB mRNA 的表达明显低于 Con 组（P=0.001、P=0.021、P=0.007），而 Uti+LPS 组 TLR4、MyD88 和 NF-κB mRNA 的表达明显高于 Uti 组（P=0.023、P=0.040、P=0.045）。
结论
这些发现表明，乌司他丁对脓毒症肠道黏膜屏障的保护作用可能是通过 TLR4/MyD88/NF-κB 通路介导的。
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