| ObjectiveChronic periodontitis is mainly treated mechanically,such as subgingival curettage.In addition,medication-assisted periodontitis is often treated.However,drug treatment mainly relies on antibiotics,which will cause problems such as dysregulation of oral flora and drug resistance.Psoralen is an effective component of psoralen,which has anti-inflammatory,antibacterial,immunomodulatory and osteogenic activities.It is widely used in the treatment of osteoporosis and shows rapid absorption in the entero-hepatic circulation.What effect does psoralen have on the intestinal tract? Does psoralen affect systemic immunity by regulating intestinal function,thus affecting bone-including alveolar bone metabolism? Based on the above background,this experiment by building the rat periodontitis model,study scurfpea fruit grain of chronic periodontitis rats alveolar bone resorption,and through the intestinal absorption of intestinal barrier,the influence of explore scurfpea fruit and its possible mechanism of effect on chronic periodontitis,for prevention and treatment of chronic periodontitis scurfpea fruit element to lay the theoretical foundation.MethodsFifty-two specific pathogen-free(SPF)SD rats,male,8 weeks old.They were randomly divided into 4 groups: control group,single administration group,model group and periodontitis rats administration group,with 13 rats in each group.Model group and periodontitis rats to medicine group rats,ligation silk ligation bilateral maxillary second molar,periodontitis model is set up,after the success of the modeling dismantling ligation silk,and at the same time on the simple drug group and periodontitis rats lavage dosage group 8 mg/Kg / 2 d scurfpea fruit element solution,the control group and model group rats,at the same frequency of physiological saline,4 w after sampling.Blood samples were taken to separate the serum,and the expression levels of inflammatory factors were detected by ELISA.The rats were sacrificed and the maxillary samples on both sides were separated.The tissue changes were observed by HE staining,the absorption value of alveolar bone was measured after micro-CT scanning and 3D reconstruction,and the expression levels of osteoclasts and osteoclasts in the alveolar bone of the maxillary second molar were detected by immunofluorescence staining.Isolation of ileal tissue and fresh intestinal contents,It was used to detect the protein expression level of NOD2 in ileal mucosa by immunohistochemical staining,the protein expression level of Occludin and Claudin-5 in ileal mucosa by ELISA,and the 16 S r RNA gene high-throughput sequencing was performed by Illumina Mi Seq platform after DNA extraction from intestinal contents.Results(1)The results of periodontal tissue detection: He staining and micro-CT 3D reconstruction showed the loss of maxillary second molar attachment,formation of periodontal pocket and resorption of alveolar bone in rats with periodontitis model.The modeling was successful.The alveolar bone loss value(P < 0.01)and the bone trabecular separation Tb.SP(P < 0.05)in the Model group were higher than those in the Peri+Pso group,while the BV/TV was lower(P < 0.05).The results indicated that psoraletin can reduce the resorption of alveolar bone caused by periodontitis.Further immunofluorescence staining showed that psoralen could decrease the RANKL/OPG ratio in rats with periodontitis.(2)Ileum and its content test results: There was no difference in the expression of Claudin-5 protein and Occludin protein.Ileal mucosal immune barrier NOD2 protein(IOD)in the Model group(P < 0.01)and Peri+Pso group(P < 0.05)was higher than that in Control group,but IOD value of Peri+Pso group was lower than that in Model group(P < 0.05).Alpha diversity analysis showed that psoralen increased Chao1 index(P <0.05)and Shannon index(P < 0.05).Further linear discriminant analysis(LEf Se)found that the relative contents of Clostridium and Clostridium increased in the Pso group.The F/B ratio in the Model group and the Pso group was lower than that in the Control group(P < 0.001).The F/B ratio in the Peri+Pso group was lower than that in the Model group(P < 0.05).(3)Results of serum inflammatory cytokines: There was no difference in the expression of IL-6.The expression level of pro-inflammatory factor TNF-α in model group was higher than that in periodontitis rats treated with psoralen group(P<0.05)and control group(P<0.01).Moreover,the expression level of IL-10 in Model group was lower than that in Peri+Pso group(P < 0.01).Covclusion(1)Psoralen can reduce alveolar bone resorption in rats with periodontitis.(2)This study demonstrated that psoralen can regulate the intestinal immune barrier and ecological barrier in rats with periodontitis.(3)Psoralen can promote the secretion of anti-inflammatory factor IL-10,reduce the secretion of pro-inflammatory factor TNF-α,and inhibit the expression of RANKL.(4)Through the above studies,we believe that whether psoralen affects the level of inflammatory factors by regulating the intestinal barrier and then reduces the resorption of alveolar bone is worthy of further study. |
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