巴戟天多糖对炎性微环境牙周膜成纤维细胞FN及FN-EDA表达的影响

doi:10.19439/j.sjos.2024.02.003

上海口腔医学 ›› 2024, Vol. 33 ›› Issue (2): 123-129.doi: 10.19439/j.sjos.2024.02.003

巴戟天多糖对炎性微环境牙周膜成纤维细胞FN及FN-EDA表达的影响

张赞¹, 戴晶怡¹, 蔡红宣¹, 司为幸¹, 杨静文^2#, 田亚光^3,4#

1.海南医学院口腔医学院,海南海口 570100;
2.北京大学口腔医院修复科,北京 100080;
3.海南医学院附属海南医院口腔科,海南海口 570100;
4.海南省人民医院口腔科,海南海口 570100

收稿日期:2023-03-28 修回日期:2023-06-06 出版日期:2024-04-25 发布日期:2024-05-14
通讯作者: 田亚光,E-mail:yaguangtian@163.com;杨静文,E-mail:jingwen.yang@foxmail.com。^#共同通信作者
作者简介:张赞（1998-）,女,硕士,E-mail: zhangzan_daisy@163.com
基金资助:
海南省重点研发计划项目（ZDYF2021SHFZ229）; 海南省自然科学基金（822CXTD534）

Effects of Morinda officinalis polysaccharides on FN and FN-EDA of periodontal ligament fibroblasts in inflammatory microenvironment

ZHANG Zan¹, DAI Jing-yi¹, CAI Hong-xuan¹, SI Wei-xing¹, YANG Jing-wen², TIAN Ya-guang^3,4

1. School of Stomatology, Hainan Medical University. Haikou 570100, Hainan Province;
2. Department of Prosthodontics, Peking University School and Hospital of Stomatology. Beijing 100080;
3. Department of Stomatology, Hainan Affiliated Hospital of Hainan Medical University. Haikou 570100, Hainan Province;
4. Department of Stomatology, Hainan General Hospital. Haikou 570100, Hainan Province, China

Received:2023-03-28 Revised:2023-06-06 Online:2024-04-25 Published:2024-05-14

摘要/Abstract

摘要： 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（10 mg/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。

关键词: 牙周炎, 巴戟天多糖, 纤连蛋白, FN-EDA

Abstract: PURPOSE: 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. 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 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. 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 expression. 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.

Key words: Periodontitis, Morinda officinalis polysaccharides, Fibronectin, Fibronectin containing extra domain A

中图分类号:

R781.4

张赞, 戴晶怡, 蔡红宣, 司为幸, 杨静文, 田亚光. 巴戟天多糖对炎性微环境牙周膜成纤维细胞FN及FN-EDA表达的影响[J]. 上海口腔医学, 2024, 33(2): 123-129.

ZHANG Zan, DAI Jing-yi, CAI Hong-xuan, SI Wei-xing, YANG Jing-wen, TIAN Ya-guang. Effects of Morinda officinalis polysaccharides on FN and FN-EDA of periodontal ligament fibroblasts in inflammatory microenvironment[J]. Shanghai Journal of Stomatology, 2024, 33(2): 123-129.

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巴戟天多糖对炎性微环境牙周膜成纤维细胞FN及FN-EDA表达的影响

Effects of Morinda officinalis polysaccharides on FN and FN-EDA of periodontal ligament fibroblasts in inflammatory microenvironment

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