【目的】 体内外观察黄芪多糖对牙周炎的治疗作用及机制。【方法】 结扎/脂多糖（LPS）诱导构建牙周炎小鼠模型，LPS处 理建立牙周炎细胞模型。给予黄芪多糖干预后，苏木精-伊红（HE）染色法检测小鼠牙周组织病理学损伤，试剂盒检测活性氧 （ROS）和丙二醛（MDA）含量及超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性等氧化应激相关指标，抗酒石酸磷酸酶 （TRAP）染色法检测牙周组织中破骨细胞的形成和RAW264.7细胞向破骨细胞分化，肌动蛋白细胞骨架/黏着斑蛋白（Vinculin）染色 法分析RAW264.7细胞中F-actin环的形成，碱性磷酸酶（ALP）染色和茜素红S（ARS）染色及ALP活性检测评价小鼠骨髓间充质 干细胞（mBMSCs）成骨细胞的形成能力，Western Blot法检测破骨细胞和成骨细胞相关蛋白的表达水平。【结果】 黄芪多糖显著 减少LPS诱导的小鼠牙槽骨流失及组织病理学损伤并改善牙周骨再生相关参数。黄芪多糖减少LPS诱导的小鼠牙周韧带细胞 （mPDLCs）中 ROS的生成，抑制 LPS处理的 mPDLCs细胞和牙周炎小鼠牙周组织和血清中 MDA含量而增加 SOD和 CAT活性。 黄芪多糖减少了LPS处理的小鼠牙周组织和RAW264.7细胞中TRAP的表达及RAW264.7细胞中F-actin环的形成。黄芪多糖降 低了LPS处理的mBMSCs细胞中 ALP的表达和活性并减少了钙沉积。此外，黄芪多糖下调了 LPS刺激的 RAW264.7细胞中破 骨细胞相关蛋白[组织蛋白酶 K（CTSK）、活化 T-细胞核因子 1（NFATc1）和 c-Fos］ 的表达并上调了 mBMSCs细胞中成骨细胞 相关蛋白[ALP、runt相关转录因子2（Runx2）、I型胶原蛋白（COL-1）和DMP1）］ 的表达。【结论】 黄芪多糖通过抑制氧化应激 及促进ERK/AMPK途径介导的骨形成能力而减轻LPS诱导的牙周炎。

Objective To observe the therapeutic effect and mechanism of astragalus polysaccharide on lipopolysaccharide（LPS）-induced periodontitis in vivo and in vitro. Methods Ligation/LPS induction was used to construct a mouse model of periodontitis，and LPS treatment was used to establish a periodontitis cellular model. After administration of astragalus polysaccharide intervention， hematoxylin-eosin （HE） staining was used to detect pathological damage in mouse periodontal tissues， and kits were used to detect reactive oxygen species （ROS） and malondialdehyde （MDA） content as well as oxidative stress-related indexes such as superoxide dismutase（SOD）and catalase（CAT）activities， and tartrate-resistant acid phosphatase（TRAP）staining was used to detect the formation of osteoclasts in periodontal tissues and the RAW264.7 cell differentiation to osteoblasts，actin cytoskeleton/focal adhesion protein（Vinculin）staining method was used to analyze the formation of F-actin ring in RAW264.7 cells，alkaline phosphatase（ALP）staining and alizarin red S（ARS）staining and ALP activity assays were performed to evaluate the osteoclast formation ability of mouse bone marrow mesenchymal stem cells（mBMSCs），and Western Blot was used to detect the expression levels of osteoclast-and osteoblastrelated proteins. Results Astragali polysaccharide significantly reduced LPS-induced alveolar bone loss and histopathological damage， as well as improved the parameters related to periodontal bone regeneration in mice.Astragalgali polysaccharide reduced ROS production in LPS-induced periodontal ligament cells （mPDLCs）， inhibited MDA content and increased SOD and CAT activities in LPS-treated mPDLCs and in periodontal tissues and serum in periodontitis mice. Astragalus polysaccharide decreased TRAP expressions in LPS-treated mouse periodontal tissues and RAW264.7 cells， and F-actin ring formation in RAW264.7 cells. Astragalgali polysaccharide decreased ALP expression and activity in LPS-treated mBMSCs cells， and reduced calcium deposition. In addition， astragalus polysaccharide down-regulated the expressions of osteoclast-related proteins [cathepsin k（CTSK）， nuclear factor of activated T-cells 1（NFATc1）and c-Fos] in LPS-inducedRAW264.7 cells，and up-regulated the expressions of osteoblast-related proteins [ALP，runt-related transcription factor 2 （Runx2）， collagen type I （ COL-1） and DMP1）] in mBMSCs. Conclusion Astragalus polysaccharide can alleviates LPS-induced periodontitis by inhibiting oxidative stress and promoting ERK/AMPK pathway-mediated bone formation capacity.

R285.5

河北省中医药类科学研究资助项目（编号：2023916）