Effects Of Grape Seed Proanthocyanidins On The Expression Of Inflammatory Mediators In Gingival Epithelial Cells

Background:Periodontitis is a complex chronic inflammatory infectious disease caused by a variety of microorganisms and the host’s immune response,mainly involving the gums,periodontal ligament,alveolar bone and other periodontal supporting tissues.Plaque microorganisms and their products act on the gums for a long time,causing the body’s immune response,and firstly leading to the inflammation of the gums.Gum inflammation caused by plaque microorganisms is considered to be a key risk factor for the onset of periodontitis.Controlling gingival inflammation is very important for preventing periodontitis.Managing gingivitis is the key strategy to prevent periodontitis,and it is also an auxiliary strategy to prevent periodontitis recurrence.Grape seed proanthocyanidins(Grape seed proanthocyanidins,GSPs),as a natural antioxidant,has multiple functions such as anti-inflammatory grade immune regulation,and it also has low toxicity.Objective:In this experiment,by studying the effects of grape seed proanthocyanidins on the proliferation activity of Human gingival epithelial cells and the inflammation induced by lipopolysaccharide,the study explored the effect of grape seed proanthocyanidins on the proliferation activity of gingival epithelial cells and the possible preventive effect on inflammation.The prevention and treatment provide a theoretical basis to promote the prevention and treatment of periodontitis with safe and efficient natural products.Method:Cell culture medium containing different concentrations of GSPs is used(0、1μg/m L 、 5μg/m L 、 10μg/m L 、 20μg/m L 、 40μg/m L 、 60μg/m L 、 80μg/m L 、100μg/m L)to treat cells in 96-well plates for 6h,12 h,24h,48 h gingival epithelial cells,and then add 100μLCulture medium and 10μLCCK-8 serum-free reagent mixture,3h later,detect the absorbance(OD value).According to the results of CCK-8,10μg/m L,20μg/m L,40μg/m L were selected to treat the cells for 24 h,and then1.0μg/m L LPS was added to stimulate 24 h,which would affect tumor necrosis factor-α(TNF-α)and interleukin-1β(interleukin-1β,IL-1β),interleukin-6(IL-6),interleukin-4(IL-4),interleukin-10(IL-10),transforming growth The content of transforming growth factor-beta(TGF-β)is detected by ELISA;Quantitative Realtime PCR(Quantitative Real-time PCR)detects TNF-α,IL-1β,IL-6,IL-4,IL-10.TGF-β gene expression level.result:1.CCK-8 results show that: compared with the control group,When the concentration of GSPs is between 0-40μg/m L,there is no significant effect on cell proliferation,and when the action time reaches 24 h,the cell proliferation rate is faster.2.ELISA and QRT-PCR results show that GSPs can effectively reduce the levels of pro-inflammatory cytokines TNF-α,IL-1β,and IL-6,while the expression of antiinflammatory cytokines IL-4,IL-10,and TGF-β raise the standard.Conclusion:When the concentration of GSPs is between 0-40μg/m L,there is no significant effect on cell proliferation,and GSPs have a significant effect on cell proliferation for24 hours.GSPs can inhibit the level of pro-inflammatory cytokines while increasing the level of anti-inflammatory cytokines,reduce the inflammatory response caused by lipopolysac-charide,and play a certain preventive effect on the resistance of gingival epithelial cells against endotoxin stimulation,and the existence of differential genes confirms GSPs.The preventive effect and the later study of the mechanism of GSPs provide a basis.