| Background: Periodontitis associated with diabetes is now recognized as the main complication of the "sixth largest" diabetes,and is the leading cause of loss of teeth in diabetic patients.Western medicine treatment of this disease is mainly based on the control of blood sugar,by removing local stimuli,and then control infection.Such as sputum scaling,subgingival scaling,root planing and other local treatment techniques,although the technical means can remove the local irritant factors such as plaque and stones attached to the tooth,but the parts that are difficult to reach the device and Invasion of pathogenic bacteria in periodontal tissues is not effective.Local or systemic application of fungicides or antibiotics to treat certain side effects of drug resistance and secondary infections,so the clinical efficacy is not good.Under the premise of removing local stimuli,the previous studies and research found that the treatment of this disease with Chinese medicine is better.Objective:To observe the appropriate concentration of Invigorating the spleen and enriching the kidney Recipe,can reduce the inflammatory cytokines,matrix metalloproteinases-2(MMP-2)in the microenvironment of human periodontal ligament cells(HPDLC)in diabetic periodontitis and the expression of the nuclear factor kappa B receptor activating factor ligand(Receptor or Activator of NF-KB Ligand,RANKL)system,in order to explore the initial mechanism of intervention of spleen and kidney to interfere with diabetic periodontitis.Method:1.The periodontal tissues were isolated in vitro,and more HPDLCs were obtained by primary and subculture.The samples were identified by strept avidin-biotin complex(SABC),and then randomly divided into experimental group and control group.The experimental group was treated with different concentrations of Buzhong Yiqi Pills and Liuwei Dihuang Pills,and the control group did not do any treatment.The cell count kit(CCK-8)was used to detect the increase in HPDLC in each group.2.HPDLC high glucose culture system was constructed by HPDLC,and randomly divided into experimental group(Liuwei Dihuang Pill Group,Buzhong Yiqi Pill Group,Liuwei Dihuang Pill + Buzhong Yiqi Pill Group)and control group(blank group,virulence factor)Group,high glucose group,high sugar + virulence factor group).The enzyme-linked immunosorbent assay(ELISA)kit was used to detect the inflammatory cytokines interleukin-1?(IL-1?)and tumor necrosis factor-?(Tumor Necrosis Factor-?)in the microenvironment,TNF-?)expression and concentration of matrix metalloproteinases-2(MMP-2).Western blotting(WB)was used to detect the expression of RANKL protein and osteoprotegerin(OPG)protein in the microenvironment.Result:1.Different concentrations of Liuwei Dihuang Pill and Buzhong Yiqi Pill have no significant effect on the proliferation of human periodontal ligament fibroblasts,the optimal concentration is 0.01mg/ml;2.Liuwei Dihuang Pill and Buzhong Yiqi Pill can reduce the expression of inflammatory cytokines IL-1? and TNF-? in the microenvironment of diabetic periodontitis;all of them can down-regulate the expression of MMP-2;there is no obvious synergistic effect;3.Liuwei Dihuang Pill,Buzhong Yiqi Pill,Liuwei Dihuang Pill and Buzhong Yiqi Pill can reduce the expression of RANKL protein in microenvironment;neither can increase the expression of OPG protein;the use of Liuwei Dihuang Pill can inhibit the RANKL system.The effect;the use of Buzhong Yiqi Pill has no inhibitory effect on the RANKL system;the combined inhibition of the RANKL system is enhanced.Conclusion:1.0.01 mg/ml of Liuwei Dihuang Pill and 0.01 mg/ml of Buzhong Yiqi Pills are more stable to human periodontal ligament fibroblasts.2.Invigorating the spleen and enriching the kidney method can reduce the expression of inflammatory cytokines IL-1? and TNF-? in the microenvironment of diabetic periodontitis;it can down-regulate the expression of MMP-2;it can inhibit the expression of RANKL system. |
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