Nuclear respiratory factor 1 mediates LPS-induced acute lung injury through NF-κB / 生理学报

The purpose of this paper was to study the transcriptional regulation of nuclear respiratory factor 1 (NRF1) on nuclear factor kappa B (NF-κB), a key molecule in lipopolysaccharide (LPS)-induced lung epithelial inflammation, and to clarify the mechanism of NRF1-mediated inflammatory response in lung epithelial cells. In vivo, male BALB/c mice were treated with NRF1 siRNA, followed with LPS (4 mg/kg) or 0.9% saline through respiratory tract, and sacrificed 48 h later. Expression levels of NRF1, NF-κB p65 and its target genes were detected by Western blot and real-time PCR. Nuclear translocation of NRF1 or p65 was measured by immunofluorescent technique. In vitro, L132 cells were transfected with NRF1 siRNA or treated with BAY 11-7082 (5 μmol/L) for 24 h, followed with treatment of 1 mg/L LPS for 6 h. Cells were lysed for detections of NRF1, NF-κB p65 and its target genes as well as the binding sites of NRF1 on RELA (encoding NF-κB p65) promoter by chromatin immunoprecipitation assay (ChIP). Results showed that LPS stimulated NRF1 and NF-κB p65. Pro-inflammatory factors including interleukin-1β (IL-1β) and IL-6 were significantly increased both in vivo and in vitro. Obvious nuclear translocations of NRF1 and p65 were observed in LPS-stimulated lung tissue. Silencing NRF1 resulted in a decrease of p65 and its target genes both in vivo and in vitro. In addition, BAY 11-7082, an inhibitor of NF-κB, significantly repressed the inflammatory responses induced by LPS without affecting NRF1 expression. Furthermore, it was proved that NRF1 had three binding sites on RELA promoter region. In summary, NRF1 is involved in LPS-mediated acute lung injury through the transcriptional regulation on NF-κB p65.

Lower expression of dynein axonemal intermediate china 1 in lung adenocarcinoma and inhibits the invasion of lung adenocarcinoma cells / 解剖学报

Objective To investigate the expression of dynein axonemal intermediate chain l(DNAI1) in lung adenocarcinoma (LUAD) and its influence on invasive ability of lung adenocarcinoma. Methods Microarray gene chip analysis was used to screen different expression genes in lung adenocarcinoma (3 samples) and adjacent normal tissues(3 samples); Heatmap and volcano plot were performed demonstrate the mRNA expression and distribution after screening; DAVID database used for Gene Ontology (GO) and Kyoto Encyclopedia of Genes of Genomes (KEGG) analysis; STRING database and Cytoscape 3.6.1 software for protein-protein interaction (PPI) analysis and screening of Hub genes; Objective genes were selected based on the differential expression of each Hub gene in lung adenocarcinoma in DEGs and Ualcan database; Real-time PCR and Western blotting were used to detect the expression of DNAI1 in BEAS-2B, H1299 and A549; observe the morphological changes after DNAI1 overexpression; Transwell invasion assay was used to detect the change of invasion ability of A549 cells after DNAI1 overexpression. Results The microarray result showed that there were 86 up-regulated genes and 396 down-regulated genes; different genes were involved in the RNA polymerase II promoter positive regulation of transcription, apoptosis process of negative regulation, protein binding, and other functions, widely distributed within the cell, and associated with the metabolic pathway, cancer and other signal pathways were closely related ; DEGs database and Ualcan database showed that DNAI1 was the most downregulated among Hub genes in LUAD; the result of Real-time PCR and Western blotting showed that DNAI1 had lower expression in H1299 and A549 compared with BEAS-2B; after DNAI1 overexpression, A549 cells became round and a few shed off; invasion assay showed that the invasion ability of A549 cells was significantly reduced. Conclusion DNAI1 has a lower expression and inhibits the ability of invasion in LUAD, and this study can provide a potential molecular target and provide a theoretical basis for targeted therapy of LUAD.

Influence of recombined human growth hormone on sIgA and EGF in rats with obstructive jaundice / 中国应用生理学杂志

<p><b>AIM</b>To study the effect of the recombined human growth hormone(rhGH) on secretory immunoglobulin A (sIgA), epidermal growth factor (EGF) in rats with obstructive jaundice.</p><p><b>METHODS</b>Sixty Wistar rats were randomly divided into four groups, sham-operated (group A), common biliary duct-ligated (group B), biliary duct-ligated plus rhGH-treat for one week (group C), biliary duct-ligated plus rhGH-treat for two weeks (group D), each group had 15 rats. Except group A, the rats of other groups were operated with biliary duct-ligated. Until two weeks after operation, the rats of group A and B were killed. After operation, the rats of group C were treated with rhGH hypodermic injection (0.75 U x kg(-1) x d(-1)) for one week, and then killed. The rats of D group were treated with rhGH hypodermic injection (0.75 U x kg(-1) x d(-1)) for two weeks, and then killed. All procedures were performed aseptically. Total bilirubin (TB), alkaline phosphatase (ALP), prealbumin(PA), insulin-like growth factor (IGF-1), sIgA, EGF were measured.</p><p><b>RESULTS</b>Compared with group A, in group B, C, D, serum level of PA, IGF-1 and sIgA, EGF level of gastric and intestinal juice were lower, but TB, ALP were higher, there were significant difference. Compared with group B, the rats with treatment of rhGH in group C and D had higher sIgA and EGF and lower intestinal bacterial translocation.</p><p><b>CONCLUSION</b>In objective jaundice rats, rhGH can protect their hepatic function, intestinal physical-barrier function and immune-barrier function, and reduce intestinal bacterial translocation.</p>