基于腹腔驻留巨噬细胞探讨扶肾方治疗腹膜透析相关性腹膜炎的机制

Exploring the mechanism of Fushen Formula in treating peritoneal dialysis-associated peritonitis based on peritoneal resident macrophages

关键词:  腹膜透析相关性腹膜炎  扶肾方  腹腔驻留巨噬细胞  氧化磷酸化  细胞因子

Key words:  peritoneal dialysis-associated peritonitis  Fushen Formula  peritoneal resident macrophages  oxidative phosphorylation  cytokines

摘要:

[目的] 基于腹腔驻留巨噬细胞（PRM）探讨扶肾方对脂多糖（LPS）诱导的腹膜透析相关性腹膜炎（PDAP）小鼠的影响及作用机制。[方法] 将36只c57BL/6J雄性小鼠按随机数字表法随机分为6组：正常组、1.5%PDS组、1.5%PDS+扶肾方低剂量（FS-L）组、1.5%PDS+扶肾方高剂量（FS-H）组、1.5%PDS+LPS组、1.5%PDS+LPS+FS-H组，每组6只。观察小鼠一般状态，采用流式细胞仪检测小鼠PRM的比例、吞噬功能及一氧化氮（NO）的生成；使用酶联免疫吸附实验（ELISA）检测各组小鼠腹腔灌洗液中白介素-6（IL-6）、白介素-10（IL-10）和肿瘤坏死因子-α（TNF-α）。[结果] 与PD组比较，LPS组小鼠PRM比例显著下降（P＜0.001），吞噬功能明显减弱（P＜0.001），NO生成显著增多（P＜0.001），腹腔灌洗液中IL-10和TNF-α也显著增多（P＜0.001）。与LPS组比较，给药小鼠PRM比例、吞噬功能均有一定的恢复（P＜0.01），NO显著下降（P＜0.001），IL-6、TNF-α显著减少（P＜0.001），IL-10明显增多（P＜0.05）。[结论] 扶肾方可以显著改善PDAP小鼠的炎症状态，可能与通过减少PRM的NO合成，促进氧化磷酸化，来调节高糖环境下腹腔的免疫稳态有关。

Abstract:

[Objective] To investigate the effects and underlying mechanisms of Fushen Formula on peritoneal dialysis-associated peritonitis（PDAP） induced by lipopolysaccharide（LPS） in mice，based on peritoneal resident macrophages（PRM）. [Methods] Thirty-six male C57BL/6J mice were randomly divided into 6 groups：the normal group，1.5% PDS group，1.5% PDS+low-dose Fushen Formula（FS-L） group，1.5% PDF+high-dose Fushen formula（FS-H） group，1.5% PDF+LPS group，1.5% PDS+LPS+FS-H group，with 6 mice in each group. The general condition of the mice was observed. The proportion of PRM，phagocytic function，and nitric oxide（NO） production were detected using flow cytometry. Levels of interleukin-6（IL-6），interleukin-10（IL-10），and tumor necrosis factor-α（TNF-α） in peritoneal lavage fluid were measured using ELISA. [Results] Compared with the PD group，the proportion of PRM in the LPS group was significantly decreased（P＜0.001），phagocytic function was markedly weakened（P＜0.001），NO production was significantly increased（P＜0.001），and levels of IL-10 and TNF-α in peritoneal lavage fluid were also significantly elevated（P＜0.001）. Compared with the LPS group，the proportion of PRM and phagocytic function in the treated mice were partially restored（P＜0.01），NO production was significantly reduced（P＜0.001），levels of IL-6 and TNF-α were significantly decreased（P＜0.001），and IL-10 was significantly increased（P＜0.05）. [Conclusion] Fushen Formula can significantly improve the inflammatory state in PDAP mice，possibly by reducing NO synthesis in PRM and promoting oxidative phosphorylation to regulate the immune homeostasis in the peritoneal cavity under high-glucose conditions.