| Periodontitis is one of the leading causes of tooth loss in adults with a high incidence,and current treatments for periodontitis aim to control the progression of the disease and reduce the risk of tooth loss.Conventional treatments for periodontitis,such as basic therapy,topical and systemic medication,and surgery,can reduce the level of periodontal bacteria.However,although traditional antibacterial drugs can improve the therapeutic effect of periodontitis,they are prone to side effects such as drug resistance and dysbacteriosis.Therefore,it is the trend of future research and development of periodontal disease to find an alternative therapy with the same source of medicine and food and no side effects.Extensive studies have proved that Moringa oleifera leaf extract can slow down the progress of various inflammatory diseases,such as colitis,nephritis,etc,and has good anti-inflammatory and antibacterial effects.The latest research suggests that cryptochlorogenic acid,an aqueous extract of Moringa oleifera leaves,has anti-inflammatory effects,but the specific mechanism is unclear.In this study,periodontitis cells and animal models were established respectively to investigate for the first time,and to explore whether cryptochlorogenic acid has anti-inflammatory and anti-bone resorption effects in experimental periodontitis,and to explore its mechanism.Objective: The anti-inflammatory and anti-bone resorption effects of cryptochlorogenic acid in experimental periodontitis and its mechanism were studied by in vitro and in vivo experiments.Methods:1.The macrophage cell line RAW264.7 was cultured,treated with different concentration gradients of cryptochlorogenic acid for 24 hours,and the cell viability was detected by CCK8.Choose the appropriate concentration of cryptochlorogenic acid and 1μg/ml LPS to co-culture for 24 h,and the expression of inflammatory factors IL-6,IL-1β,TNF-a,IL-10 and the bone protection system RANKL/OPG were detected by qRT-PCR.Western blot was used to detect the expression levels of the bone protection system RANKL/OPG and p-p38/p38 in each group.2.An animal model of periodontitis was established,divided into 5 groups:blank control group,periodontitis group,cryptochlorogenic acid group,p38 MAPK inhibitor SB203580 group,cryptochlorogenic acid+p38 MAPK inhibitor SB203580 group.After 1 month of execution,HE staining detects the toxic effect of the drug on each organ.Micro-CT scanning measures alveolar bone resorption,and qRT-PCR detects expression of inflammatory factors.The Western blot method detected the expression levels of the bone protection system RANKL/OPG and p-p38/p38 in each group.Results:1.In vitro and in vitro experiments found that cryptochlorogenic acid had little effect on RAW264.7 cells and experimental rats within the appropriate concentration range.2.Cryptochlorogenic acid can inhibit the expression of pro-inflammatory factors IL-6,IL-1β and TNF-a,and up-regulate the expression of anti-inflammatory factor IL-10 in vitro and in vivo,and high concentration of cryptochlorogenic acid has better anti-inflammatory effect.3.Different concentrations of cryptochlorogenic acid can inhibit the expression of RANKL/OPG in vitro and in vivo.Micro-CT scan of rat alveolar bone found that cryptochlorogenic acid can reduce the resorption of alveolar bone and has a bone protection effect.4.Both in vitro and in vivo experiments showed that cryptochlorogenic acid can down-regulate the expression of p-p38/p38,and the anti-inflammatory and bone protective effects of cryptochlorogenic acid may be related to the inhibition of p38 MAPK signaling pathway activation.Conclusions: Cryptochlorogenic acid has anti-inflammatory and anti-bone resorption effects,and its effects may be related to the inhibition of p38 MAPK signaling pathway activation. |
