Naringin attenuates MLC phosphorylation and NF-κB activation to protect sepsis-induced intestinal injury via RhoA/ROCK pathway
Naringin attenuates MLC phosphorylation and NF-κB activation to protect sepsis-induced intestinal injury via RhoA/ROCK pathway
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DOI：
10.1016/j.biopha.2018.03.163
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发表时间：
2018-07-01
影响因子：
7.5
通讯作者：
Yang, Mingshi
中科院分区：
文献类型：
--
作者：
Li, Zhiling;Gao, Ming;Yang, Mingshi
背景：脓毒症通常与宿主免疫系统的过度刺激有关，导致多器官功能衰竭。据报道，柚皮苷具有多种生物效应。本研究旨在探讨柚皮苷对脓毒症引起的肠屏障功能损伤的体内外保护作用。方法：将小鼠随机分为4组，分别为假手术组（n=20）、CLP+药组（n=20）、CLP+NG （30 mg/kg）组（n=20）和CLP+NG （60 mg/kg）组（n=20）。盲肠结扎穿刺（CLP）致脓毒症。采用H&E染色、透射电镜观察肠黏膜形态。采用ELISA法测定大鼠体内外肠道通透性和炎症反应。Western blot和RhoA活性测定测定紧密连接蛋白水平和指示信号通路的激活。MTT法测定细胞活力。结果：柚皮苷提高了CLP小鼠的存活率，减轻了败血症引起的肠黏膜损伤。此外，柚皮苷改善了受损的肠通透性，抑制了tnf - α和IL-6的释放，同时增加了CLP小鼠和脂多糖（LPS）刺激的MODE-K细胞中IL-10的水平，并呈剂量依赖性。柚皮苷在体内外通过RhoA/ROCK/NF-kappa B/MLCK/MLC信号通路增加紧密连接蛋白ZO-1和clclin -1的表达。结论：柚皮苷在体内外通过RhoA/ROCK/NF-kappa B/MLCK/MLC信号通路改善脓毒症诱导的肠道损伤。
Background: Sepsis is commonly associated with excessive stimulation of host immune system and result in multi-organ failure dysfunction. Naringin has been reported to exhibit a variety of biological effects. The present study aimed to investigate the protective effect of naringin on sepsis-induced injury of intestinal barrier function in vivo and in vitro.Methods: Mice were randomly divided into 4 groups named sham (n=20), CLP + vehicle (n=20), CLP+NG (30 mg/kg) (n=20) and CLP+NG (60 mg/kg) (n=20) groups. Sepsis was induced by cecal ligation and puncture (CLP). H&E staining and transmission electron microscopy (TEM) were performed to observe intestinal mucosal morphology. ELISA was used to determine the intestinal permeability and inflammatory response in vivo and in vitro. Western blot and RhoA activity assay were performed to determine the levels of tight junction proteins and the activation of indicated signaling pathways. MTT assay was used to determine cell viability.Results: Naringin improved survival rate of CLP mice and alleviated sepsis-induced intestinal mucosal injury. Furthermore, naringin improved impaired intestinal permeability and inhibited the release of TNF-alpha and IL-6, while increased IL-10 level in CLP mice and lipopolysaccharide (LPS)-stimulated MODE-K cells in a dose-dependent manner. Naringin increased the expression of tight junction proteins ZO-1 and claudin-1 via RhoA/ROCK/NF-kappa B/MLCK/MLC signaling pathway in vivo and in vitro.Conclusion: Naringin improved sepsis-induced intestinal injury via RhoA/ROCK/NF-kappa B/MLCK/MLC signaling pathway in vivo and in vitro.