醒脑静注射液活性成分对氧糖剥夺/复氧损伤的PC12细胞的保护作用研究

Protection of active pharmaceutical ingredients in the Xingnaojing Injection against oxygen glucose deprivation/re-oxygenation-induced PC12 cells injury

目的研究醒脑静注射液中8种活性成分(麝香酮、吉马酮、莪术二酮、莪术烯醇、β-榄香烯、龙脑、樟脑和天然冰片)对氧糖剥夺/复氧(OGD/R)损伤的PC12细胞的保护作用。方法将PC12细胞分为10组:正常对照组、模型组和醒脑静8种活性成分分别给药组。正常对照组正常培养,模型组OGD 6 h/R 18 h,给药组在OGD 6 h/R 18 h时分别给予各活性成分10μmol·L^-1。用比色法检测细胞活力、乳酸脱氢酶(LDH)释放量以及抗氧化酶(SOD和CAT)的活力。结果正常对照组、模型组、吉马酮组和龙脑组的CAT活力分别为1.00±0.00,0.81±0.14,1.30±0.35和1.08±0.16;正常对照组、模型组、麝香酮组、吉马酮组、龙脑组和天然冰片组的LDH释放量分别为1.00±0.00,2.14±0.17,1.86±0.15,1.85±0.12,1.86±0.10和1.83±0.14;正常对照组、模型组、吉马酮组、莪术二酮组、龙脑组、樟脑组和天然冰片组的细胞活力分别为1.00±0.00,0.60±0.07,0.75±0.08,0.72±0.06,0.75±0.08,0.79±0.10和0.79±0.12;正常对照组、模型组、麝香酮组、吉马酮组、莪术二酮组、天然冰片组、莪术烯醇组、β-榄香烯组、龙脑组以及樟脑组SOD的活力分别为1.00±0.00,1.27±0.30,4.78±1.36,5.12±1.26,2.62±0.28,3.06±0.46,4.31±1.45,3.34±0.93,4.77±1.85和3.87±1.39。上述指标:模型组与正常对照组相比,差异均有统计学意义(P<0.05,P<0.01);4个给药组、6个给药组、7个给药组和10个给药组分别与模型组相比,差异均有统计学意义(均P<0.05)。结论醒脑静注射液中的吉马酮、龙脑、樟脑和天然冰片均能通过调控氧化应激发挥抗OGD/R损伤的作用。

Objective To observe the effect of eight active pharmaceutical ingredients in Xingnaojing Injection,which include Muscone,Germacrone,Curdione,Curcumenol,β-elemene,Borneolum,Camphor,and d-Borneolum,on oxygen glucose deprivation/re-oxygenation(OGD/R)-induced PC12 cells.Methods PC12 cells were divided into 10 groups:Normal control group,model group,and eight compounds individually treated groups.The cells in normal control group were cultured normally.The cells in model group were treated with OGD 6 h/R 18 h.The administration groups was treated with 10μmol·L^-1 of each active ingredient at the same time as OGD 6 h/R 18 h.Cell viability,lactate dehydrogenase(LDH)release and antioxidant enzymes(SOD and CAT)activities were measured by colorimetric method.Results The CAT activities in normal control group,model group,Germacrone group,and Borneolum group were 1.00±0.00,0.81±0.14,1.30±0.35,1.08±0.16;the LDH release in normal control group,model group,Muscone group,Germacrone group,Borneolum group,and d-Borneolum group were 1.00±0.00,2.14±0.17,1.86±0.15,1.85±0.12,1.86±0.10 and 1.83±0.14;the cell viability rates in normal control group,model group,Gemimarone group,Curcumenol group,Borneolum group,Camphor group,and d-Borneolum group were 1.00±0.00,0.60±0.07,0.75±0.08,0.72±0.06,0.75±0.08,0.79±0.10 and 0.79±0.12;the SOD activities in normal control group,model group,Muscone group,Germacrone group,Curdione group,d-Borneolum group,Curcumenol group,β-elemene group,Borneolum group,and Camphor group were 1.00±0.00,1.27±0.30,4.78±1.36,5.12±1.26,2.62±0.28,3.06±0.46,4.31±1.45,3.34±0.93,4.77±1.85 and 3.87±1.39.Comparison between model group and normal control group,the difference of the factors were significant(P<0.05,P<0.01).Comparison between four administration groups or five administration groups,or eight administration groups,or ten administration groups and model group,respectively,the differences of the factors were significant(all P<0.05).Conclusion In Xingnaojing Injection,Germacrone,Borneolum,Camphor,and d-Borneolum could protect PC12 cell from OGD/R-induced injury by regulating oxidative stress,which may be the material basis for Xingnaojing Injection to play a neuroprotective role.