ABSTRACT
Periodontitis is a chronic inflammatory disease induced by Porphyromonas gingivalis (P. gingivalis) and other pathogens. P. gingivalis release various virulence factors including lipopolysaccharide (LPS). However, whether P. gingivalis-LPS inducing pyroptosis in human gingival fibroblasts (HGFs) remains unknown. In present study, P. gingivalis-LPS decreased the membrane integrity of HGFs, and pyroptosis-associated cytokines were upregulated at the mRNA level. In addition, pyroptosis proteins were highly expressed in gingival tissues of periodontitis. P. gingivalis-LPS induced gingivitis in the rat model, and the expression level of pyroptosis-associated proteins increased. Together, P. gingivalis-LPS can activate the pyroptosis reaction, which may be a pro-pyroptosis status in a relative low concentration.
Subject(s)
Fibroblasts/drug effects , Gingiva/drug effects , Gingivitis/chemically induced , Lipopolysaccharides/toxicity , Porphyromonas gingivalis/metabolism , Pyroptosis/drug effects , Animals , Caspase 1/metabolism , Cells, Cultured , Cytokines/genetics , Cytokines/metabolism , Disease Models, Animal , Female , Fibroblasts/metabolism , Fibroblasts/pathology , Gingiva/metabolism , Gingiva/pathology , Gingivitis/metabolism , Gingivitis/pathology , Humans , Lipopolysaccharides/isolation & purification , Male , Rats, Sprague-Dawley , Signal Transduction , Up-RegulationABSTRACT
OBJECTIVE: To establish a capillary electrophoresis-mass spectrometry(CE-MS) method for the analysis of nineteen organonitrogen pesticides in Paeoniae Radix Alba. METHODS: CE-MS analysis was performed on a 70 cm X 50 µm fused-silica capillary. The optimal buffer was composed of 1 % formic acid and 15 % methanol(V/V, pH 2.2). The temperature of capillary was controlled at 25 degree. The separation voltage was +20 kV. The optimal MS parameters were as follows: ESI-MS analysis was performed in the positive mode; 90 % methanol containing 0.2 % formic acid with a flow rate of 8 µl·min(-1) was selected as the sheath liquid; the flow rate and temperature of drying gas were 6 L·min(-1) and 250 degree, respectively; The nebulizing gas pressure was set at 5 psig; The optimal values of fragmentor and ESI voltage were 100 V and 5 000 V, respectively. RESULTS: The nineteen pesticides had good linearity over the testing ranges. The average recoveries were in the range of 80.1 %-108.4 % with RSDs less than 20 % (except ethoxyquin and spiroxamine, those of which were 29.2 % and 22.3 % at 0.01 mg.kg(-1) concentration level). The LODs of nineteen pesticides were 0.503 â10.1 µg.kg(-1). CONCLUSION: The method can be used effectively to analyze the nineteen organonitrogen pesticides residue in Paeoniae Radix Alba.
