Zhang Zan, Dai Jing-Yi, Cai Hong-Xuan, Si Wei-Xing, Yang Jing-Wen, Tian Ya-Guang
School of Stomatology, Hainan Medical University. Haikou 570100, Hainan Province,China. E-mail:
Shanghai Kou Qiang Yi Xue. 2024 Apr;33(2):123-129.
To investigate the effect of Morinda officinalis polysaccharides(MOP) on the expression of fibronectin(FN) and fibronectin containing extra domain A(FN-EDA) in inflammatory periodontal ligament fibroblasts.
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 confirmed by Micro-CT to complete the modeling. The remaining rats in the model group were randomly divided into periodontitis 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 injected, 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 detected by immunohistochemistry. Periodontal ligament fibroblasts were cultured in vitro, the effect of MOP on cell activity detected by CCK-8. The fourth generation cells were divided into control group, inflammation group (10 mg/mL lipopolysaccharide), and experimental group (12.5 μg/mL MOP, 12.5 μg/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 software package.
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 expression.
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.
探讨巴戟天多糖(MOP)对炎性牙周膜成纤维细胞中纤连蛋白(FN)及含A额外结构域的纤连蛋白(FN-EDA)表达的影响。
将36只大鼠随机分为对照组(n=12)和模型组(n=24)。模型组采用正畸钢丝结扎法建立牙周炎模型。3周后,每组选取6只大鼠,通过Micro-CT确认建模成功。模型组剩余大鼠随机分为牙周炎组、生理盐水(NS)组和MOP组。MOP组大鼠左侧上颌第一磨牙腭侧注射MOP(200 mg/kg,连续3天,4周内每天50 μL)。NS组注射等量NS,牙周炎组不做处理。取大鼠左侧上颌组织,通过苏木精-伊红(H-E)染色观察牙周组织的病理变化。采用免疫组织化学法检测FN和FN-EDA的表达。体外培养牙周膜成纤维细胞,通过CCK-8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(10 mg/mL脂多糖)和实验组(12.5 μg/mL MOP、12.5 μg/mL MOP+10 mg/mL脂多糖)。采用实时定量聚合酶链反应(qRT-PCR)和蛋白质免疫印迹法(Western blot)检测FN和FN-EDA的表达。数据采用Prism 8.0软件包进行统计学分析。
体内实验中,MOP组FN-EDA的表达明显低于牙周炎组和NS组(P<0.05),且炎性细胞浸润减少。然而,各组FN表达无显著差异。体外实验中,与对照组相比,炎症组FN-EDA mRNA和蛋白表达显著增加(P<0.000 1)。MOP显著降低炎性细胞中FN-EDA的表达,但对FN表达无显著影响。
随着炎性牙周膜组织和细胞中FN-EDA表达增加,MOP可能通过下调FN-EDA发挥抗炎作用。