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Background@#As Actinobacillus pleuropneumonniae (APP) infection causes considerable losses in the pig industry, there is a growing need to develop effective therapeutic interventions that leverage host immune defense mechanisms to combat these pathogens. @*Objectives@#To demonstrate the role of microRNA (miR)-127 in controlling bacterial infection against APP. Moreover, to investigate a signaling pathway in macrophages that controls the production of anti-microbial peptides. @*Methods@#Firstly, we evaluated the effect of miR-127 on APP-infected pigs by cell count/ enzyme-linked immunosorbent assay (ELISA). Then the impact of miR-127 on immune cells was detected. The cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 were evaluated by ELISA. The expression of cytokines (anti-microbial peptides [AMPs]) was assessed using quantitative polymerase chain reaction. The expression level of IL-6, TNF-α and p-P65 were analyzed by western blot. The expression of p65 in the immune cells was investigated by immunofluorescence. @*Results@#miR-127 showed a protective effect on APP-infected macrophage. Moreover, the protective effect might depend on its regulation of macrophage bactericidal activity and the generation of IL-22, IL-17 and AMPs by targeting sphingosine-1-phosphate receptor3 (SIPR3), the element involved in the Toll-like receptor (TLR) cascades. @*Conclusions@#Together, we identify that miR-127 is a regulator of S1PR3 and then regulates TLRuclear factor-κB signaling in macrophages with anti-bacterial acticity, and it might be a potential target for treating inflammatory diseases caused by APP.
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OBJECTIVE:To establish HPLC fingerprints of Potentilla discolor,and to conduct authenticity identification.METHODS:HPLC method was adopted.The determination was performed on InertSustain C18 column with mobile phase consisted of 0.1％ formic acid solution-acetonitrile (gradient elution) at the flow rate of 1.0 mL/min,detection wavelength of 360 nm,colunn temperature of 30 ℃,sample size of 10 μL.Using rutin as reference,HPLC chromatograms of 19 batches of P.discolor and 2 batches of P.chinesis were determined.TCM Fingerprint Similarity Evaluation System (2004) was used for similarity evaluation of 21 batches of samples,and common peak identification of 19 batches of P discolor SPSS 21.0 statisticl software was used for main component analysis and cluster analysis.RESULTS:There were 18 common peaks in HPLC fingerprints of 19 batches of P.discolor,the similarity was higher than 0.9.-HPLC chromatogram was in good agreement with control fingerprint.The similarity of 2 batches of P chinesis was lower than 0.7.The 21 batches of medicinal materials could be grouped into 2 categories,2 batches of P chinesis could be grouped into a category,19 batches of P.discolor could be grouped into a category.P discolor could be grouped into 4 categories.Rutin and quercitrin were main ingredients in 19 batches of P discolor.CONCLUSIONS:Established fingerprint can provide reference for authenticity identification and quality evaluation of P.discolor.
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AIM:To determine whether caudatin , a C21 steroidal aglycone , enhances tumor necrosis factor-re-lated apoptosis-inducing ligand ( TRAIL)-associated HepG2 cell apoptosis .METHODS:Cell growth inhibition was deter-mined by MTT assay and cell colony formation assay .The TUNEL apoptosis detection kit was used to analyze cell apopto-sis, and the protein expression was examined by Western blotting .RESULTS:Combination of caudatin with TRAIL signi-ficantly reduced cell proliferation and increased the apoptotic rate of HepG 2 cells compared with the use of each agent alone.This was evidenced by marked increases in caspase-3, caspase-7, caspase-9 and PARP cleavages in the cells treated with caudatin and TRAIL-compared with control group .Combination of caudatin with TRAIL also led to the strong suppres-sion of survivin .CONCLUSION:Caudatin synergizes HepG 2 cells to TRAIL-induced apoptosis by promoting the cleava-ges of caspase-3, caspase-7, caspase-9 and PARP and inhibiting the expression of survivin .