醒脑静注射液活性成分麝香酮对LPS诱导的BV-2小胶质细胞炎症反应的影响

Effect of Xingnaojing Injection's active ingredient muscone on LPS-induced BV-2 microglia inflammatory response

目的筛选醒脑静注射液8种单体(吉马酮、莪术二酮、β-榄香烯、樟脑、莪术烯醇、麝香酮、天然冰片、龙脑)中抑制BV-2细胞炎症反应的活性成分,研究其对炎症因子释放的影响。方法CCK-8法检测8种单体(10μmol/L)对小胶质细胞活力的影响;10μmol/L的8种单体孵育BV-2细胞0.5 h,用0.1μg/mL脂多糖(LPS)进行刺激,培养24 h后收集上清,Greiss法检测NO浓度;Elisa检测肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)的浓度。不同浓度的麝香酮孵育BV-2细胞0.5 h,用0.1μg/mL LPS刺激,培养24 h后收集上清,Greiss法检测NO浓度;Elisa法检测TNF-α、IL-6、白介素1受体α(IL-1Rα)的浓度。结果与对照组比较,醒脑静注射液各个单体成分对正常培养的BV-2细胞无毒性作用。LPS刺激BV-2细胞后,模型组与对照组比较,产生的NO、TNF-α、IL-6、IL-1Rα显著增多(P<0.01);与模型组比较,麝香酮5.0、7.5、10.0μmol/L浓度可显著抑制NO、IL-6的产生,10μmol/L麝香酮可显著抑制TNF-α的产生,差异均有统计学意义(P<0.01);其他7种单体成分均无显著影响;2.5、5.0μmol/L麝香酮组IL-1Ra的释放量有升高趋势,但无统计学意义。结论麝香酮可以显著抑制LPS诱导的BV-2细胞炎性因子的产生。

Objective To study the effect of 8 monomers of Xingnaojing injection(gematrone,zedoary diketone,β-elemene,camphor,zedoary enol,musk ketone,borneol,borneol)on the release of inflammatory factors,the active components were screened.Methods CCK-8 was used to detect the effects of eight monomers on the microglia activity.Xingnaojing eight monomers of 10μmol/L concentration:gemone,sputum dione,β-elemene,camphor,zephyrenol,musk ketone,natural borneol,borneol to incubate BV-2 cells for 0.5 h,then stimulated with LPS.The supernatant was collected after 24 h of culture.The Greens method was used to detect the concentration of NO.Elisa detected tumor necrosis factor(TNF-α)and the concentration of interleukin-6(IL-6).Different concentrations of musk ketone were used to incubate BV-2 cells for 0.5 h,then stimulated with lipopolysaccharide.After 24 h of culture,the supernatant was collected.CCK-8 was used to detect the effects of different concentrations of musk ketone on the microglia viability.Greiss method NO concentration was measured;Elisa detected the concentrations of tumor necrosis factor(TNF-α),interleukin 6(IL-6),and interleukin-1 receptor alpha(IL-1Rα).Results Compared with the control group,Xingnaojing injection has no toxic effect on BV-2 cells.After LPS stimulated BV-2 cells,NO,TNF-α,IL-6 and IL-1Rαwere significantly increased in the model group compared with the control group(P<0.01).Compared with the model group,musk ketone of 5.0,7.5,and 10.0μmol/L can significantly inhibit the production of NO and IL-6,the difference was statistically significant(P<0.01).The other 7 monomers had no significant effect.The release of IL-1Ra in 10μmol/L musk ketone group increased,but there was no statistical significance.Conclusion Musk ketone can inhibit the inflammatory activation of LPS-induced microglia and inhibit the production of inflammatory factors.