Let-7b通过靶向P38丝裂原活化蛋白激酶对闭合蛋白表达的保护作用在预防肠道屏障功能障碍中的研究
Protective Effects of Let-7b on the Expression of Occludin by Targeting P38 MAPK in Preventing Intestinal Barrier Dysfunction.
作者信息
Liu Zhihua, Tian Yinghai, Jiang Yanqiong, Chen Shihua, Liu Ting, Moyer Mary Pat, Qin Huanlong, Zhou Xinke
机构信息
Department of AnoRectal Surgery, the Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
Department of Center Laboratory, the Fifth Affiliated Hospital of Guangzhou Medical University, Guangzhou, China.
出版信息
Cell Physiol Biochem. 2018;45(1):343-355. doi: 10.1159/000486815. Epub 2018 Jan 19.
BACKGROUND/AIMS: Let-7b was dramatically reduced after a dicer knockout of mice with intestinal barrier function injuries. This paper aims to investigate the molecular mechanism of let-7b by targeting p38 MAPK in preventing intestinal barrier dysfunction.
METHODS
A total of 186 patients were enrolled, with 93 in the control group and 93 in the PRO group. Only 158 patients completed the entire study, whereas the others either did not meet the inclusion criteria or refused to participate. To further verify the role of let-7b, intestinal epithelial conditional knockout (IKO) mice of mmu-let-7b model were established. Serum let-7b, zonulin, IL-6, and TNF-α concentrations were measured by ELISA or quantitative RT-PCR. Permeability assay was done by ussing chamber. The apoptotic cells were identified using an In Situ Cell Death Detection Kit. Protein was detected by western blot.
RESULTS
Probiotics can lower infection-related complications, as well as increase the serum and tissue let-7b levels. P38 MAPK was identified as the target of let-7b, as verified by NCM460 cells. P38 MAPK expression was increased, whereas tight-junction (TJ) proteins were significantly decreased in let-7b IKO mice (both P<0.05). Negative regulation of p38 MAPK molecular signaling pathways was involved in the protective effects of let-7b on intestinal barrier function.
CONCLUSION
Let-7b was identified as a novel diagnosis biomarker or a potential treatment target for preventing intestinal barrier dysfunction.
背景/目的：在肠道屏障功能受损的dicer基因敲除小鼠中，Let-7b显著降低。本文旨在研究Let-7b通过靶向p38丝裂原活化蛋白激酶（p38 MAPK）预防肠道屏障功能障碍的分子机制。
方法
共纳入186例患者，对照组93例，益生菌组（PRO组）93例。仅158例患者完成了整个研究，其他患者要么不符合纳入标准，要么拒绝参与。为进一步验证Let-7b的作用，建立了mmu-Let-7b模型的肠道上皮条件性敲除（IKO）小鼠。采用酶联免疫吸附测定（ELISA）或定量逆转录聚合酶链反应（RT-PCR）检测血清Let-7b、zonulin、白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）浓度。使用小室进行通透性测定。使用原位细胞死亡检测试剂盒鉴定凋亡细胞。通过蛋白质印迹法检测蛋白质。
结果
益生菌可降低感染相关并发症，并提高血清和组织中Let-7b水平。经NCM460细胞验证，p38 MAPK被确定为Let-7b的靶点。在Let-7b IKO小鼠中，p38 MAPK表达增加，而紧密连接（TJ）蛋白显著降低（均P<0.05）。p38 MAPK分子信号通路的负调控参与了Let-7b对肠道屏障功能的保护作用。
结论
Let-7b被确定为预防肠道屏障功能障碍的新型诊断生物标志物或潜在治疗靶点。
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