益生菌联合消退素D1对牙周炎大鼠牙龈组织中炎症因子、正五聚体蛋白3和氧化应激的影响

Effects of Probiotics Combined with Resolvin D1 on Inflammatory Factors Pentraxin 3 and Oxidative Stress in the Gingival Tissue of Rats with Periodontitis

目的:研究益生菌联合消退素D1对牙周炎大鼠牙龈组织中炎症因子、正五聚体蛋白3(Pentraxin 3,PTX3)和氧化应激的影响。方法:将大鼠分成对照组、模型组(牙周炎大鼠)、益生菌组(牙周炎大鼠,益生菌治疗)、消退素D1组(牙周炎大鼠,消退素D1治疗)、联合组(牙周炎大鼠,益生菌联合消退素D1治疗),检测牙龈出血指数、菌斑指数及探诊深度,ELISA法检测牙龈组织中白细胞介素-1β(Interleukin-1β,IL-1β)、白细胞介素-6(Interleukin-6,IL-6)和肿瘤坏死因子α(Tumor necrosis factor,TNF-α)水平,Western blot检测牙龈组织中PTX3、核因子-κB(Nuclear factor-κB,NF-κB)p65、p-p38丝裂原活化蛋白激酶(Mitogen-activated protein kinases,MAPK)、基质金属蛋白酶-9(Matrix metalloprotease 9,MMP-9)和基质金属蛋白酶-2(Matrix metalloprotease 2,MMP-2)蛋白表达,比色法检测牙龈组织中超氧化物歧化酶(Superoxide orgotein dismutase,SOD)、丙二醛(Malondialdehyde,MDA)和谷胱甘肽过氧化物酶(Glutathione peroxidase,GSH-Px)含量。结果:与对照组比较,模型组大鼠牙龈出血指数、菌斑指数、探诊深度均升高,炎症因子IL-1β、IL-6、TNF-α含量升高,PTX3、NF-κBp65、p-p38MAPK、MMP-9、MMP-2蛋白表达量升高,SOD、GSH-Px含量降低,MDA含量升高,差异有统计学意义(P<0.05)。与模型组比较,益生菌组、消退素D1组、联合组大鼠牙龈出血指数、菌斑指数、探诊深度均降低,炎症因子IL-1β、IL-6、TNF-α含量降低,PTX3、NF-κBp65、p-p38MAPK、MMP-9、MMP-2蛋白表达量降低,SOD、GSH-Px含量升高,MDA含量降低,差异有统计学意义(P<0.05)。与益生菌组、消退素D1组比较,联合组大鼠牙龈出血指数、菌斑指数、探诊深度均降低,炎症因子IL-1β、IL-6、TNF-α含量降低,PTX3、NF-κBp65、p-p38MAPK、MMP-9、MMP-2蛋白表达量降低,SOD、GSH-Px含量升高,MDA含量降低,差异有统计学意义(P<0.05)。结论:益生菌联合消退素D1可抑制牙周炎大鼠牙龈组织中炎症因子表达,抑制氧化应激,机制可能与抑制p38MAPK、NF-κB信号转导有关。

Objective To study the effect of probiotics combined with resolvin D1 on inflammatory factors,pentrameric protein 3(PTX3)and oxidative stress in the gingival tissue of rats with periodontitis.Methods Rats were divided into control group,model group(periodontitis rats),probiotic group(periodontitis rats,probiotic treatment),resolvin D1 group(periodontitis rats,resolvin D1 treatment),combined group(periodontitis rats,treated with probiotics and resolvin D1).Gingival bleeding index,plaque index and probing depth were measured.The levels of IL-1β,IL-6 and TNF-αin gingival tissue were detected by ELISA.The levels of PTX3,NF-κBp65,MAPK,MMP-9 and MMP-2 in gingival tissue were detected by Western blot.The levels of SOD,MDA,GSH-Px in gingival tissue were detected by colorimetric method.Results Compared with the control group,the gingival bleeding index,plaque index and probing depth of rats in the model group increased,and the levels of inflammatory factors IL-1β,IL-6 and TNF-αincreased,PTX3,NF-κBp65,p-p38MAPK,MMP-9 and MMP-2 protein expression increased,SOD and GSH-Px decreased,MDA increased,the differences were statistically significant(P<0.05).Compared with the model group,the gingival bleeding index,plaque index,and probing depth of rats in the the probiotic group,resolvin D1 group and the combination group decreased,and the levels of inflammatory factors IL-1β,IL-6 and TNF-αdecreased,and PTX3,NF-κBp65,MAPK,MMP-9 and MMP-2 protein expression decreased,SOD and GSH-Px content increased,MDA decreased(P<0.05).Compared with the probiotic group and the resolvin D1 group,the gingival bleeding index,plaque index,and probing depth of rats in the combined group decreased,and the levels of inflammatory factors IL-1β,IL-6 and TNF-αdecreased,and PTX3,NF-κBp65,MAPK,MMP-9 and MMP-2 protein expression decreased,SOD and GSH-Px content increased,MDA decreased(P<0.05).Conclusion The combination of probiotics and resolvin D1 inhibits the expression of inflammatory factors in the gingival tissue of rats with periodontitis and inhibits oxidative stress,the mechanism may be related to the inhibition of p38MAPK and NF-κB signal transduction.