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ObjectiveTo investigate the effect of Xuanfei Zhisou prescription on the interleukin-17 （IL-17） signaling pathway in model rats with chronic obstructive pulmonary disease （COPD）. MethodA total of 60 Wistar rats were randomly divided into a blank group （10 rats） and a model group （50 rats）， and COPD model rats were established by tracheal infusion of lipopolysaccharide combined with passive fumigation. After modeling， the rats were divided into the model group， dexamethasone group， and high， medium， and low-dose Xuanfei Zhisou prescription groups （3.6， 1.8， 0.9 g·kg-1·d-1） according to the random number table. Rats in the blank group and model group were given normal saline of 10 mL·kg-1·d-1 by gavage administration， and the intervention groups of Xuanfei Zhisou prescription were given corresponding drugs. Rats in the dexamethasone group were given dexamethasone of 2.57×10-4 g·kg-1·d-1 for 28 days. The level of pulmonary function indexes in rats was measured by a pulmonary function detector. The contents of interleukin-6 （IL-6）， interleukin-8 （IL-8）， IL-17， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） were detected by enzyme-linked immunosorbent assay （ELISA）. The positive expressions of IL-17A， IL-17RA， nuclear factor-κB activator 1 （Act1）， tumor necrosis factor-associated factor 6 （TRAF6）， p-p38 mitogen-activated protein kinase （p-p38 MAPK）， nuclear factor-κB p65 （NF-κB p65）， and phosphorylation were detected by immunohistochemistry （IHC）. The protein expression levels of IL-17A， IL-17RA， Act1， and TRAF6 in the lung tissue were detected by Western blot. The mRNA expressions of IL-17A， IL-17RA， Act1， and TRAF6 in the lung tissue were detected by Real-time polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the serum contents of IL-6， IL-8， IL-17， IL-1β， and TNF-α in the model group were significantly increased （P<0.05）， and the flow rate and volume indexes of pulmonary function in the model group were significantly decreased （P<0.05）， while the time indexes and other indexes were significantly increased （P<0.05）. The mRNA and protein expression levels of IL-17A， IL-17RA， Act1， and TRAF6 in pulmonary tissue and the positive expressions of downstream NF-κB p65， p-NF-κB p65， and p-p38 MAPK were increased （P<0.05）. Compared with the model group， the levels of IL-6， IL-8， IL-17， IL-1β， and TNF-α in the serum of all treatment groups were decreased to varying degrees （P<0.05）， and the indexes of pulmonary function were improved to different degrees （P<0.05）. The mRNA and protein levels of IL-17A， IL-17RA， Act1， and TRAF6 and the positive expression of downstream NF-κB p65， p-NF-κB p65， and p-p38 MAPK in high and medium-dose Xuanfei Zhisou prescription groups were significantly decreased （P<0.05）. ConclusionXuanfei Zhisou prescription can effectively resist inflammation of COPD rats. The mechanism may be related to down-regulating the protein expression of IL-17A， IL-17RA， Act1， and TRAF6， inhibiting downstream NF-κB and p38 MAPK signaling pathways， and reducing the release of IL-6， IL-8， TNF-α， IL-17， and IL-1β， thus reducing the airway inflammation response.
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Objective@# To investigate the transcriptomic changes in primary human oral keratinocytes (pHOKs) after coculture with Prevotella melaninogenica (P.m) and to verify the changes in human oral keratinocyte (HOK) cell lines.@*Methods@# pHOK was isolated and cocultured with P.m for 0, 4 and 24 h. Total RNA was extracted, a gene library was constructed, transcriptional sequencing was performed, differentially expressed genes (DEGs) were analyzed, gene ontology (GO) pathway analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed, and the validation of DEGs was performed by qRT-PCR and Western Blot in the HOK and P.m coculture cell model. @*Results @#1 788 DEGs were detected between the 4 h group and control group, including upregulated DEGs such as lymphocyte cytosolic protein 1(LCP1), keratin 7 (KRT7) and Cilia and flagella associated protein 251(CFAP251) and downregulated DEGs such as FERM, ARH/RhoGEF and Pleckstrin domain protein 1 (FARP1), WW domain containing transcription regulator 1(WWTR1) and Discoidin, CUB and LCCL domain-containing protein 2 (DCBLD2). 1 832 DEGs were detected between the 24 h group and control group, including upregulated DEGs such as LCP1, complement C1s(C1S), kynureninase (KYNU) and downregulated DEGs such as phosphoserine aminotransferase 1 (PSAT1), FARP1 and FKBP prolyl isomerase 10 (FKBP10). There were 1 090 common differentially expressed genes (cDEGs) in the 4 h and 24 h groups, including LCP1, KYNU and long intergenic nonprotein coding RNA 958 (LINC00958). The GO pathways were mainly enriched in response to lipopolysaccharide and the molecules of bacterial origin and apical part of the cell. KEGG pathway analysis revealed enrichment in the interleukin-17 (IL-17) signaling pathway, tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) pathway, etc. We verified the expression of a cDEG, Myosin1B (MYO1B), and qRT-PCR and Western Blot analysis showed that MYO1B expression was significantly upregulated between the control group and the P.m cocultured group (P<0.001), and its expression followed a time-dependent and concentration-dependent manner. @*Conclusion@#P.m played an important role in the transcriptome of oral keratinocytes.