RESUMO
To investigate the effect of prophylactic aucubin (AU) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice. Methods: Male BABL/c mice were randomly divided into a control group, an ALI group, and an AU treatment group, 16 mice in each group. ALI mice were injected with LPS (5 mg/kg, intratracheal injection), and AU (10 mg/kg) was injected intraperitoneally 30 min ahead. After LPS injection for 6 hours mice were sacrificed, the morphological changes of lung tissues were detected by HE staining and the lung injury score was obtained. The mRNA expression of tumor necrosis factor-α (TNF-α) and interleukin 10 (IL-10) in lung tissue was detected by real-time PCR. The total protein and lactate dehydrogenase (LDH) activity, the cell count, and the protein content of TNF-α and IL-10 in the mouse bronchoalveolar lavage fluid (BALF) were detected. Results: Compared with ALI mice, the pathological damage score of lung tissue was significantly reduced in the AU group, the total number of BALF cells, neutrophils, and macrophages were significantly decreased, LDH activity and the total protein content were also significantly decreased (all P<0.01). In addition, AU can reduce the mRNA and protein expression of TNF-α in lung of ALI mice, and increase the mRNA and protein expression of IL-10 (all P<0.01). Conclusion: AU can reduce LPS-induced ALI in mice.
Assuntos
Animais , Masculino , Camundongos , Lesão Pulmonar Aguda , Líquido da Lavagem Broncoalveolar , Glucosídeos Iridoides , Lipopolissacarídeos , Pulmão , Fator de Necrose Tumoral alfaRESUMO
Objective To clone,express and purify thioredoxin (named as N5),a specific diagnostic antigen of hepatitis E virus (HEV),and to initially evaluate its antigenicity and serological test performance.Methods Based on the gene sequences of HEV-ORF2 and carboxyl terminal ORF3 on GenBank,the codon was optimized by the Escherichia coli codon preference,inserted it into prokaryotic expression vector M48 following total gene synthesization,and expressed in Escherichia coli fusion protein N5 recombined with Thioredoxin (TRX).Fusion protein was purified in affinity chromatography,evaluating its antigenicity with Western blot technology,then evaluating its serological test performance using the negative and positive serum samples confirmed of HEV infection with laboratory and clinical tests.Results The recombinant plasmid expressing N5 diagnostic antigen was successfully established; high-level expression and purification to obtain soluble diagnostic antigens; Western blot results indicating fusion protein N5 can be bound specifically with the serum of HEV IgM antibody positive,showing satisfactory antigencity; using fusion protein N5 as the capture antigen to build indirect ELISA,testing 40 serum samples of HEV cases confirmed by pathogen detection and clinical diagnosis and 40 serum samples of healthy people,with the sensitivity and specificity of 95％ (38/40) and 90％ (36/40) respectively.Conclusion Recombinant plasmid expressing the HEV diagnostic antigen recombined with thioredoxin was successfully established,and soluble fusion protein N5 was obtained with high expression and strong antigenicity,promising in its future applications.
RESUMO
Objective To determine the effect of calcitonin gene-related peptide (CGRP) on triggering receptor expressed on myeloid cells-1 (TREM-1) in the lipopolysaccharide (LPS)-induced macrophages and its signal transduction pathway. Methods The levels of TREM-1 mRNA in the macrophages were observed by reverse transcription-polymerase chain reaction (RT-PCR), and flow cytometry was performed to detect TREM-1 protein expression levels in the macrophages. Results CGRP had no regulating effect on the expression of TREM-1 in the macrophages; LPS could up-regulate macrophages to express TREM-1; CGRP increased TREM-1 mRNA expression in LPS-induced macrophages in dose and time-dependent manner; CGRP increased TREM-1 protein expression in LPS-induced macrophages, which could be partially reversed by H-7 or H-89 (P<0.05). Conclusion CGRP can regulate the LPS-induced macrophages synthesis and secretion of TREM-1, and the intracellular signal transduction pathway is related to PKA and PKC.