[关键词]
[摘要]
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(10 μmol/L)对小胶质细胞活力的影响;10 μmol/L的8种单体孵育BV-2细胞0.5 h,用0.1 μg/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0.1 μg/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、10.0 μmol/L浓度可显著抑制NO、IL-6的产生,10 μmol/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、5.0 μmol/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。
[Key word]
[Abstract]
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 10 μmol/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-1R α were 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 10.0 μmol/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 10 μmol/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.
[中图分类号]
R285.5
[基金项目]
国家自然科学基金资助项目(81573644);国家中药标准化项目(ZYBZH-C-JS-35);“十三五”期间天津市高等学校“创新团队培养计划”(NO.TD13-5050)