Effects of Morinda officinalis polysaccharides on FN and FN-EDA of periodontal ligament fibroblasts in inflam-matory microenvironment
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目的:探讨巴戟天多糖(morinda officinalis polysaccharides,MOP)对炎性微环境牙周膜成纤维细胞纤连蛋白(fibronectin,FN)及含额外结构域 A 的纤连蛋白(fibronectin containing extra domain A,FN-EDA)表达的影响.方法:将36只大鼠随机分为对照组(n=12)和模型组(n=24),模型组采用正畸丝结扎法建立牙周炎,3周后每组各取6只大鼠通过Micro-CT确认建模完成;剩余模型组大鼠随机分为牙周炎组、生理盐水组(NS组)和MOP组.MOP组于大鼠左侧上颌第一磨牙腭侧注射MOP(200 mg/kg、3 d,50 µL、4周),NS组注射等体积NS,牙周炎组不做任何处理.取大鼠左侧上颌骨组织,采用H-E染色观察牙周组织病理改变,免疫组织化学染色检测FN、FN-EDA的表达.体外培养牙周膜成纤维细胞,CCK-8法检测MOP对细胞活性的影响.将第4代细胞分为对照组、炎症组(10mg/mL脂多糖)及实验组(12.5 μg/mL MOP,12.5 μg/mL MOP+10 mg/mL 脂多糖).利用 qRT-PCR 及 Western 印迹法检测 FN 和 FN-EDA 的表达变化.采用Prism 8.0软件包对数据进行统计学分析.结果:体内实验中,MOP组较牙周炎组和NS组FN-EDA表达显著下降(P＜0.05),炎细胞浸润减少;但各组FN表达量未见统计学差异.体外实验中,与对照组相比,炎症组FN-EDA mRNA和蛋白表达量显著增高(P＜0.000 1);MOP显著降低炎症细胞FN-EDA表达,但对FN表达无明显影响.结论:FN-EDA在炎症牙周膜组织和细胞中表达升高,MOP可能通过下调FN-EDA而发挥炎症抑制作用.
PURPOSE:To investigate the effect of Morinda officinalis polysaccharides(MOP)on the expression of fi-bronectin(FN)and fibronectin containing extra domain A(FN-EDA)in inflammatory periodontal ligament fibroblasts.METHODS:Thirty six rats were randomly divided into a control group(n=12)and a model group(n=24).The model group used orthodontic wire ligation to establish periodontitis.After three weeks,6 rats from each group were selected and con-firmed by Micro-CT to complete the modeling.The remaining rats in the model group were randomly divided into peri-odontitis group,normal saline(NS)group,and MOP group.In the MOP group,MOP(200 mg/kg for 3 d,50 µL for 4 weeks)was injected into the palatal side of the left maxillary first molar of the rats.In the NS group,same volume of NS was in-jected,and no treatment was performed in the periodontitis group.The left maxillary tissue of rats were taken and the pathological changes of periodontal tissue were observed by H-E staining.The expression of FN and FN-EDA was detect-ed by immunohistochemistry.Periodontal ligament fibroblasts were cultured in vitro,the effect of MOP on cell activity de-tected by CCK-8.The fourth generation cells were divided into control group,inflammation group(10 mg/mL lipopolysac-charide),and experimental group(12.5 μg/mL MOP,12.5 pg/mL MOP+10 mg/mL lipopolysaccharide).The expression of FN and FN-EDA was detected by qRT-PCR and Western blot.The data were statistically analyzed using Prism 8.0 soft-ware package.RESULTS:In vivo experiments,the expression of FN-EDA in the MOP group was significantly lower than that in the periodontitis group and NS group(P＜0.05),and the infiltration of inflammatory cells was reduced.However,there was no significant difference in the expression of FN in each group.In vitro experiments,compared with the control group,the expression of FN-EDA mRNA and protein in the inflammation group was significantly increased(P＜0.000 1).MOP significantly reduced the expression of FN-EDA in inflammatory cells,but had no significant effect on FN expres-sion.CONCLUSIONS:With increased expression of FN-EDA in inflammatory periodontal ligament tissues and cells,MOP may play a role in inhibiting inflammation by down-regulating FN-EDA.
PeriodontitisMorinda officinalis polysaccharidesFibronectinFibronectin containing extra domain A
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