芍药苷对Aβ_(1-42)诱导的小胶质细胞炎性反应和趋化性的影响

Effect of paeoniflorin on the Aβ_(1-42)-induced inflammation and chemotaxis of microglia

目的探讨芍药苷(PF)对β-淀粉样蛋白(Aβ)_(1-42)诱导的小胶质细胞炎性反应和趋化性的影响。方法原代培养大鼠小胶质细胞,分别用0、5、25、50μmol/LPF预处理细胞后,采用CCK-8法和LDH法检测细胞毒性。将培养的细胞分为空白对照组(正常培养基)、阳性对照组(Aβ_(1-42)处理细胞)和实验组(PF+Aβ_(1-42)处理细胞)。采用ELISA法检测各组肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)和趋化因子配体1(CXCL1)、趋化因子配体2(CCL2)的表达水平。采用Transwell小室进行小胶质细胞体外趋化实验,观察各组细胞趋化迁移情况。结论培养大鼠原代小胶质细胞纯度达90%以上,CCK-8法和LDH法测定均显示PF对小胶质细胞无明显细胞毒作用(均P>0.05)。PF可抑制Aβ_(1-42)诱导的小胶质细胞分泌促炎性介质TNF-α、IL-1β、IL-6和趋化因子CXCL1、CCL2(均P<0.01)。同时,PF可抑制小胶质细胞向Aβ_(1-42)的趋化(均P<0.05)。结论 PF可抑制Aβ_(1-42)诱导的啮齿动物小胶质细胞生成促炎性介质和趋化因子,并抑制小胶质细胞向Aβ_(1-42)趋化迁移,提示PF可能为阿尔茨海默病的治疗提供新的契机。

Objective To investigate the effect of paeoniflorin （PF） on the beta-amyloid （Aβ）1-42-induced inflammation and chemotaxis of microglia.Methods Primary cultures of rat microglia were pretreated with 0, 5, 25, 50 μmol/L PF respectively.To determine the cytotoxic effect of PF on microglia, CCK-8 assay was performed, and the activity of LDH in culture supernatant was detected.Microglia were divided into three groups： a blank control group （normal culture medium）, a positive control group （Aβ1-42-treated cells） and an experimental group （PF-pretreated and then Aβ1-42-treated cells）.Levels of tumor necrosis factor （TNF）-α, interleukin （IL）-1β, IL-6, chemokine （C-X-C motif） ligand 1 （CXCL1） and chemokine （C-C motif） ligand 2 （CCL2） in the culture supernatant were determined by ELISA kits.Transwell inserts were used to determine the chemotactic migration of microglia in vitro.Results The purity of cultured microglia was more than 90%.PF exerted no significant cytotoxic effect on microglia viability according to CCK-8 assay and LDH assay, no statistical difference between groups （P〉0.05） was found.PF suppressed Aβ1-42-induced secretion of proinflammatory mediators （TNF-α, IL-1β and IL-6） and chemokines （CXCL1 and CCL2） from microglia between the experimental group and the positive control group（P〈0.01）.And PF inhibited microglial chemotaxis towards Aβ1-42 （P〈0.05） between the experimental group and the positive control group in the number of migrated cells.Conclusions The above findings suggest that PF reduce Aβ1-42-stimulated production of proinflammatory mediators and chemotactic factors from rodent microglia, as well as inhibit microglial chemotaxis towards Aβ1-42.This suggests that PF might provide a potential new therapeutic strategy for Alzheimer＆#39;＆#39;s disease.