摘要
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.
出处
《中国神经免疫学和神经病学杂志》
CAS
2017年第4期276-280,289,共6页
Chinese Journal of Neuroimmunology and Neurology
基金
国家自然科学基金资助项目(81300939)
辽宁省科学技术计划项目(2009225027)
辽宁省自然科学基金项目(20170541036)