人参皂苷Rb3对大鼠实验性牙周炎的疗效研究

doi:10.7518/hxkq.2025.2024448

摘要/Abstract

摘要：

目的探讨人参皂苷Rb3对大鼠实验性牙周炎炎症及骨吸收的疗效及作用机制。方法体内实验将雄性SD大鼠随机分为对照组、结扎组、Rb3组和多西环素（Dox）组，采用上颌第二磨牙结扎法建立牙周炎模型，药物处理3周处死取材。显微计算机断层扫描（Micro-CT）评估牙槽骨吸收情况；苏木精-伊红（HE）染色观察牙周组织和内脏组织病理学改变；抗酒石酸酸性磷酸酶（TRAP）染色检测牙周组织破骨细胞形成；酶联免疫吸附试验（ELISA）检测血清白细胞介素（IL）-6、IL-8、免疫球蛋白（Ig）M、IgG水平；定量聚合酶链式反应（qPCR）检测牙龈组织炎症和破骨形成相关因子表达；免疫荧光染色检测磷酸化胞外信号调节激酶（p-ERK）表达；体外实验将RAW264.7细胞药物预处理后加入牙龈卟啉单胞菌（P.gingivalis）脂多糖（LPS）刺激。qPCR法检测IL-1β、IL-6 mRNA的表达；蛋白质免疫印迹（Western blot）检测Rb3对丝裂原活化蛋白激酶（MAPKs）信号通路的影响。结果与对照组相比，结扎组大鼠牙周炎症及骨吸收明显；与结扎组相比，Rb3组大鼠牙槽骨吸收和破骨细胞形成数量减少，牙周组织p-ERK/ERK比值、IL-1β、IL-6、活化T细胞核因子（NFATc1）mRNA及破骨下游基因表达降低、血清IL-6、IL-8、IgG、IgM的含量减少；Rb3降低P.gingivalis LPS刺激RAW264.7细胞引起的IL-8、IL-1β mRNA表达及p-ERK/ERK、p-p38 MAPK/p38 MAPK比值。结论 Rb3抑制大鼠实验性牙周炎炎症及骨吸收，且相较于Dox，Rb3在抑制促炎症因子、破骨基因表达等方面效果更佳，并可能通过激活MAPKs信号通路发挥抗炎作用。

关键词: 人参皂苷Rb3, 实验性牙周炎, 炎症, 骨吸收, 丝裂原活化蛋白激酶信号通路

Abstract:

Objective This study aimed to explore the therapeutic effect and mechanism of ginsenoside Rb3 on experimental periodontitis and bone resorption in rats. Methods Male SD rats were randomly divided into a control group, a ligation group, an Rb3 group, and a doxycycline (Dox) group for in vivo experiments. A periodontitis model was established by ligating the maxillary second molar, and samples were collected after 3 weeks of drug treatment. Micro-CT assessment of alveolar bone resorption was performed, and hematoxylin-eosin (HE) staining was used to observe pathological changes in periodontal and visceral tissues. Tartrate resistant acid phosphatase (TRAP) staining was applied to detect the formation of osteoclasts in periodontal tissues, and enzyme-linked immunosorbent assay (ELISA) was adopted to detect the serum levels of interleukin (IL)-6, IL-8, immunoglobulin (Ig)M, and IgG. Quantitative polymerase chain reaction (qPCR) was employed to detect the expression of factors related to gingival inflammation and osteoclast formation. Immunofluorescence staining was used to detect phospho-extracellular signal-regulated kinase (p-ERK) expression. In vitro experiments were conducted by pretreating RAW264.7 cells with drugs and adding lipopolysaccharides (LPS) stimulation from Porphyromonas gingivalis (P. gingivalis). IL-1β and IL-6 mRNA expression was detected by qPCR, and Western blot was used to detect the effect of Rb3 on the mitogen-activated protein kinases (MAPKs) signaling pathway. Results Compared with the control group, the ligation group showed significant periodontitis and bone resorption. Compared with the ligation group, the Rb3 group showed a decrease in alveolar bone resorption and osteoclast formation; p-ERK/ERK ratio, IL-1β, IL-6, and nuclear factor of activated T cells (NFATc1) mRNA levels and downstream gene expression in periodontal tissues; serum IL-6, IL-8, IgG, and IgM levels. Rb3 reduced IL-8 and IL-1β mRNA expression levels and p-ERK/ERK and p-p38 MAPK/p38 MAPK ratios in RAW264.7 cells induced by P. gingivalis LPS stimulation. Conclusion Rb3 inhibits inflammation and bone resorption in experimental periodontitis in rats. Compared with Dox, Rb3 has better effects in inhibiting pro-inflammatory factors and osteoclast gene expression and may exert anti-inflammatory effects by activating the MAPK signaling pathway.

Key words: ginsenoside Rb3, experimental periodontitis, inflammation, bone resorption, mitogen-activated protein kinases signaling pathway

中图分类号:

R781.4

李华, 张康, 曲会娟, 冀洪海, 孙敏敏. 人参皂苷Rb3对大鼠实验性牙周炎的疗效研究[J]. 华西口腔医学杂志, 2025, 43(5): 711-721.

Li Hua, Zhang Kang, Qu Huijuan, Ji Honghai, Sun Minmin. Effect of ginsenoside Rb3 on experimental periodontitis in rats[J]. West China Journal of Stomatology, 2025, 43(5): 711-721.

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引物	序列（5’-3’）	扩增长度/bp
GAPDH	F：GACATGCCGCCTGGAGAAAC	92
	R：AGCCCAGGATGCCCTTTAGT
IL-6	F：ACTTCCAGCCAGTTGCCTTCTTG	110
	R：TGGTCTGTTGTGGGTGGTATCCTC
IL-1β	F：AATCTCACAGCAGCATCTCGACAAG	98
	R：TCCACGGGCAAGACATAGGTAGC
CTSK	F：TGGCTGTGGAGGCGGCTATATG	82
	R：CGGGTAAGCGTCTTCAGAGTCAATG
ACP5	F：ATGACGCCAATGACAAGAGGTTCC	125
	R：TTGTGCCGAGACATTGCCAAGG
NFATc1	F：TCCCTGCCTGCCCTTGACTG	92
	R：TGAGCCCTGTGGTGAGACTTGG
MMP-9	F：CTCCTGGTGCTCCTGGCTCTAG	112
	R：GCTGTGTGTCCGTGAGGTTGG

引物	序列（5’-3’）	扩增长度/bp
GAPDH	F：GGTTGTCTCCTGCGACTTCA	183
	R：TGGTCCAGGGTTTCTTACTCC
IL-8	F：GAGACTTCCATCCAGTTGCCTTC	151
	R：TGTGTAATTAAGCCTCCGACTTGTG
IL-1β	F：TCGCAGCAGCACATCAACAAG	190
	R：CCAGCAGGTTATCATCATCATCCC