ABSTRACT
OBJECTIVE@#To construct a NF-kappaB siRNA expression vector and to detect the specific silencing effect of the siRNA on the expression of NF-kappaB protein.@*METHODS@#pcDNA3.1/CT-GFP-TOPO recombinant eukaryotic expression vector and pSilencer 1.0-U6-siRNA-NF-kappaB recombinant vector were constructed respectively. These 2 recombination plasmids were co-transfected into COS-7 cells, and the NF-kappaB silence induced by RNAi was detected by Western blot and the inverted fluorescence microscope.@*RESULTS@#The levels of NF-kappaB protein in COS-7 cells could be silenced effectively and specifically by pSilencer 1.0-U6-siRNA- NF-kappa recombinant vector. The expression of NF-kappaB protein was reduced gradually with the increase of pSilencer 1.0-U6-siRNA- NF-kappaB recombinant vector,which could be detected by Western blot under the inverted fluorescence microscope.@*CONCLUSION@#NF-kappaB siRNA expression vector is constructed successfully.
Subject(s)
Animals , COS Cells , Chlorocebus aethiops , Genetic Vectors , NF-kappa B , Genetics , RNA Interference , RNA, Small Interfering , TransfectionABSTRACT
OBJECTIVE@#To observe the effects of xipayi mouth rinse of different concentrations on the activities of interleukin-6 (IL-6) from human gingival fibroblast (HGF) induced by lipopolysaccharide (LPS).@*METHODS@#HGF was stimulated with LPS at 25 g/mL, and the radioimmunoassay (RIA) was used to examine the effect of xipayi mouth rinse at 12.5 approximately 200 g/mL on the secretion of IL-6 in the supernatant of the cell culture.@*RESULTS@#IL-6 secreted by human gingival fibroblast was significantly inhibited by xipayi mouth rinse in a dose dependent manner.@*CONCLUSION@#Xipayi mouth rinse can inhibit the secretion of IL-6 from HGF induced by LPS, suggesting the anti-inflammatory effect of xipayi mouth rinse to treat and prevent periodontal diseases.
Subject(s)
Humans , Anti-Inflammatory Agents , Pharmacology , Cells, Cultured , Depression, Chemical , Drugs, Chinese Herbal , Pharmacology , Fibroblasts , Metabolism , Gingiva , Cell Biology , Interleukin-6 , Metabolism , Lipopolysaccharides , MouthwashesABSTRACT
OBJECTIVE@#To observe the effects of xipayi mouth rinse on the DNA synthesis and change of cell cycles of human gingival fibroblast (HGF) induced by lipopolysaccharide(LPS).@*METHODS@#HGF was stimulated with LPS at 25 mg/L. Flow cytometry was used to examine the effect of xipayi mouth rinse at 25 mg/L on the DNA synthesis and change of HGF cell cycles.@*RESULTS@#The percentage of HGF in G( 1) phase increased after the cells were induced by LPS, while the percentage of HGF in S phase decreased. Xipayi mouth rinse could ameliorate this phenomenon.@*CONCLUSION@#Xipayi mouth rinse can significantly ameliorate the inhibitory effect of LPS on the proliferation of HGF, suggesting the anti-inflammatory effect of xipayi mouth rinse in the treatment and prevention of periodontal diseases.