宾达利对脂多糖诱导的BV-2细胞炎症因子释放和吞噬作用的影响

Effect of bindarit on the lipopolysaccharide-induced release of inflammatory cytokines and phagocytosis in BV-2 microglial cells

目的探讨单核细胞趋化蛋白1(MCP-1)抑制药宾达利对脂多糖(LPS)诱导的小鼠BV-2小胶质细胞炎症因子表达及吞噬作用的影响。方法将BV-2细胞分为5组,空白组不给予药物处理,正常培养36 h;模型组正常培养12 h后,加入500 ng·mL^(-1)LPS继续作用24 h;低、中、高剂量实验组分别给予20,40,80μg·mL^(-1)宾达利处理12 h后,加入500 ng·mL^(-1)LPS继续作用24 h。用CCK-8法检测各组BV-2细胞存活率,用激光共聚焦显微镜观察BV-2细胞对荧光微球的吞噬能力的改变,用实时荧光定量聚合酶链反应法检测单核细胞趋化蛋白1(MCP-1)和白细胞介素-6(IL-6)mRNA的表达水平。结果低、高剂量实验组和模型组、空白组的细胞存活率分别为(104.90±1.90)%,(100.30±4.71)%,(105.30±3.56)%和(100.00±9.44)%,各组间的细胞存活率比较,差异均无统计学意义(均P>0.05)。低、高剂量实验组和模型组、空白组的BV-2细胞所吞噬的荧光微球数量分别为(77.67±8.39),(50.00±14.73),(118.30±26.76)和(56.67±4.73)个,MCP-1 mRNA分别为2.52±0.47,1.31±0.24,2.48±0.11和1.00±0.18,IL-6 mRNA分别为18.96±1.92,8.85±0.43,31.96±4.08和1.00±0.08。高剂量实验组的上述指标与模型组比较,差异均有统计学意义(均P<0.05)。结论宾达利可能通过降低MCP-1水平、抑制IL-6分泌,进而改善LPS诱导的小胶质细胞异常吞噬作用。

Objective To investigate the effect of bindarit,an inhibitor of monocyte chemotactic protein 1 (MCP-1),on the lipopolysaccharide(LPS)-induced release of inflammatory cytokines and phagocytosis in mouse BV-2 microglial cells.Methods BV-2 cells were divided into5 groups.The blank group was not treated with drugs and cultured normally for 36 h;the model group was cultured normally for 12 h and then added 500 ng·m L;LPS for 24 h;the experimental-L,-M,-H groups were treated with 20,40,80μg·m L;bindarit for 12 h and then added 500 ng·m L;LPS for 24 h.The cell viability of BV-2microglial cells was measured with CCK-8.The phagocytosis of BV-2microglial cells was observed by laser confocal microscopy.The expression levels of monocyte chemotactic protein 1 (MCP-1) and interleukin-6 (IL-6) mRNA were detected by reverse transcriptionpolymerase chain reaction.Results The cell viabilities of experimental-L,-H groups,model group and blank group were (104.90±1.90)%,(100.30±4.71)%,(105.30±3.56)%and (100.00±9.44)%,respectively.There were no significant differences among all groups (P>0.05).The number of fluorescent microspheres phagocyted by BV-2 cells in experimental-L,-H groups,model group and blank group was (77.67±8.39),(50.00±14.73),(118.30±26.76) and (56.67±4.73),MCP-1 mRNA was 2.52±0.47,1.31±0.24,2.48±0.11 and 1.00±0.18,IL-6 mRNA was 18.96±1.92,8.85±0.43,31.96±4.08 and 1.00±0.08.There were statistically significant differences between model group and experimental-H group (all P<0.05).Conclusion Bindarit may improve the abnormal phagocytosis of microglia induced by LPS through reducing the level of MCP-1and inhibiting the secretion of IL-6.