P物质在牙周炎诱导全身炎症反应致多器官功能损伤中的作用和机制研究

Role and mechanism of substance P in multi-organ dysfunction induced by systemic inflammatory response caused by periodontitis

目的本研究通过在牙周炎模型小鼠和体外细胞实验中给予NK-1受体拮抗剂阻断P物质(SP),探讨SP在牙周炎诱导全身炎症反应致多脏器功能损伤中的作用。方法通过反复龈沟注射牙龈卟啉单胞菌脂多糖(LPS)的方式诱导牙周炎小鼠模型。同时给予NK-1受体拮抗剂,观察牙周炎小鼠全身炎症因子分泌及器官功能损伤情况;细胞实验使用LPS和SP分别刺激巨噬细胞,酶联免疫吸附实验(ELISA)测定上清液中的IL-1β,加入NK-1受体拮抗剂,观察上清液中IL-1β的水平。结果与对照组相比,LPS诱导的牙周炎组龈沟液中、血清中的SP含量均明显升高(P<0.001),同时血清中IL-1β、IL-6、TNF-α的水平也较对照组升高(P<0.001)。LPS组的谷丙转氨酶(ALT)和谷草转氨酶(AST)轻度升高(P<0.05),肺组织病理学评分有显著性差异(P<0.001)。给予NK-1受体拮抗剂,血清中IL-1β水平较LPS组降低,差异有统计学意义(P<0.05),且小鼠ALT的水平降低,肺组织病理学评分改善。NK-1受体拮抗剂能够部分逆转SP诱导巨噬细胞产生IL-1β的作用,有统计学差异(P<0.001)。结论SP通过与NK-1受体结合参与了牙周炎诱导全身炎症反应致多器官功能损伤。

Objective In this study,NK-1 receptor antagonist was given to block the function of substance P(SP)in periodontitis model mice and cell experiments in vitro to explore the role of SP in multi-organ dysfunction induced by systemic inflammatory response caused by periodontitis.Methods Periodontitis mouse model was induced by repeated gingival sulcus injection of Porphyromonas gingivalis lipopolysaccharides(LPS).At the same time,NK-1 receptor antagonist was given to observe the secretion of systemic inflammatory factors and functional damage of various organs in periodontitis mice;macrophages were stimulated by LPS and SP respectively,and IL-1β in the supernatant was monitored by enzyme-linked immunosorbent assay(ELISA);NK-1 receptor antagonist was added to observe the IL-1β level in supernatant.Results Compared with the control group,SP in gingival crevicular fluid and serum in LPS induced periodontitis group were significantly increased(P<0.001),and the level of IL-1β,IL-6 and TNF-αin serum was also significantly increased(P<0.001).Alanine aminotransferase(ALT)and aspartate aminotransferase(AST)increased slightly in LPS group(P<0.05),and there was significant difference in lung histopathological score(P<0.001).NK-1 receptor antagonist decreased IL-1β in serum,and the level of ALT was significantly lower than that in LPS group(P<0.05).NK-1 receptor antagonist could partially reverse the production of IL-1β secreted by macrophages induced by SP(P<0.001).Conclusion SP participates in the appearance of multi-organ dysfunction induced by systemic inflammatory response caused by periodontitis through binding with NK-1 receptor.