Hong Linliang, Huang Huanhuan, Wu Bin
Department of Pediatrics, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China.
Exp Ther Med. 2021 Sep;22(3):999. doi: 10.3892/etm.2021.10431. Epub 2021 Jul 15.
Intestinal microfloras are involved in various types of cancer; however, there is a limited amount of research into the involvement of metabolites of intestinal microflora (MIM) in asthmatic airway epithelial cells (AECs). The present study was designed to reveal the functions and mechanisms of MIM in the asthmatic inflammation of AECs. House dust mite (HDM)-induced asthma cell models were established and treated with mouse MIM. A MTT assay was used to investigate AEC viability, while reverse transcription-quantitative PCR and western blot analysis were used to measure the expression levels of miR-200c-3p, IL6ST, JNK and STAT3 in asthmatic AECs. ELISA was used to measure the concentration of IL-5 and IL-6. Furthermore, the targeting relationship between microRNA(miR)-200c-3p and IL6ST was investigated using a luciferase reporter gene assay. Compared with normal human bronchial epithelial cells, HDM-induced AECs had lower expression level of miR-200c-3p, higher mRNA and protein expression levels of IL6ST and an increase in IL-5 and IL-6 concentration. Both MIM and miR-200c-3p mimics suppressed the secretion of IL-5 and L-6 and promoted the proliferation of HDM-induced AECs. MIM could also upregulate miR-200c-3p and downregulate IL6ST and proteins in the JNK/STAT3 pathway. IL6ST was found to be a downstream target of miR-200c-3p. Inhibition of miR-200c-3p reversed the suppression of asthmatic inflammation by MIM. In summary, MIM upregulated miR-200c-3p expression level to reduce the protein and mRNA expression levels of IL6ST and suppress its downstream JNK/STAT3 signaling pathway, therefore inhibiting the asthmatic inflammation of AECs.
肠道微生物群与多种类型的癌症有关；然而，关于肠道微生物群代谢产物（MIM）在哮喘气道上皮细胞（AECs）中的作用的研究却很有限。本研究旨在揭示MIM在AECs哮喘炎症中的功能和机制。建立了屋尘螨（HDM）诱导的哮喘细胞模型并用小鼠MIM进行处理。采用MTT法检测AECs的活力，同时采用逆转录定量PCR和蛋白质印迹分析检测哮喘AECs中miR-200c-3p、IL6ST、JNK和STAT3的表达水平。采用ELISA法检测IL-5和IL-6的浓度。此外，使用荧光素酶报告基因检测法研究微小RNA（miR）-200c-3p与IL6ST之间的靶向关系。与正常人支气管上皮细胞相比，HDM诱导的AECs中miR-200c-3p的表达水平较低，IL6ST的mRNA和蛋白表达水平较高，且IL-5和IL-6浓度升高。MIM和miR-200c-3p模拟物均抑制了IL-5和IL-6的分泌，并促进了HDM诱导的AECs的增殖。MIM还可上调miR-200c-3p并下调JNK/STAT3通路中的IL6ST和蛋白。发现IL6ST是miR-200c-3p的下游靶点。抑制miR-200c-3p可逆转MIM对哮喘炎症的抑制作用。总之，MIM上调miR-200c-3p的表达水平，以降低IL6ST的蛋白和mRNA表达水平，并抑制其下游的JNK/STAT3信号通路，从而抑制AECs的哮喘炎症。
