通塞脉制剂对过氧化氢致PC12细胞损伤的保护作用  

The Protective Effect of Tongsaimai Decoction on PC12 Cells Induced by H_2O_2

作　　者：李锦吉 狄留庆[1,2] 李俊松[1,2] 毕肖琳 

机构地区：[1]南京中医药大学药学院,江苏南京210023 [2]江苏省中药高效给药系统工程技术研究中心,江苏南京210023

出　　处：《南京中医药大学学报》2014年第2期146-149,共4页Journal of Nanjing University of Traditional Chinese Medicine

基　　金：国家自然科学基金(81073071)

摘　　要：目的 观察通塞脉制剂对过氧化氢（H2O2）作用下PC12细胞的保护作用.方法 体外培养PC12细胞,将细胞分为正常对照组,过氧化氢组,通塞脉高剂量组（5 mg/mL）,通塞脉中剂量组（0.5 mg/mL）,通塞脉低剂量组（0.05 mg/mL）.200μmol/L过氧化氢被加入作为刺激物孵化4h后,再加入终浓度5、0.5、0.05 mg/mL的药物,继续培养24 h,用MTT法测定并计算药物对H2O2致PC12细胞损伤的保护率;取各组细胞上清液,按照试剂盒的操作方式,测定超氧化物歧化酶（SOD）,丙二醛（MDA）,一氧化氮（NO）,乳酸脱氢酶（LDH）及钙离子含量（Ca2＋）.结果 通塞脉制剂（5、0.5、0.05 mg/mL）对过氧化氢致PC12细胞损伤均有保护作用.过氧化氢组细胞上清液中SOD活性,SOD抑制率降低（P＜0.05～0.01）,MDA,LDH,NO含量与Ca2＋含量增加（P＜0.05～0.01）.而各给药组能提高SOD活性及抑制率,降低LDH,MDA,NO以及Ca2＋含量,与过氧化氢组比较有显著性差异（P＜0.05～0.01）.结论 通塞脉制剂对PC12细胞损伤有显著性保护作用,其机制与钙超载,氧化应激等因素有关.OBJECTIVE To observe the protective effects of Tongsaimai decoction preparation on the PC12 cells induced by H2O2.METHODS The in-vitro cultured PC12 cells were randomly divided into eight groups such as normal group,hydrogen peroxide group,Tongsaimai decoction with high dose group（5 mg/mL）,Tongsaimai decoction with middle dose group（0.5 mg/mL）,Tongsaimai decoction with low dose group（0.05 mg/mL）.After the 200μmol/L H2O2 was added to stimulate the PC12 cells and incubated for 4 h,then the final concentration of drugs （5 mg/mL,0.5 mg/mL,0.05 mg/mL） were added and continuously incubated and cultured for 24 h.The MTT assay was facilitated to measure and calculate the cell viability the protection rate of the drugs on PC 12 cells induced by hydrogen peroxide.Finally,the corresponding biochemical indicators like SOD,MDA,NO,LDH and Ca2＋ were measured.RESULTS The Tongsaimai decoction（5 mg/mL,0.5 mg/mL,0.05 mg/ mL） possess protective effects on H2O2-cellular injury model group induced to PC12 cells injury.The hydrogen peroxide model group revealed that could reduce the SOD activity and SOD inhibition rate （P ＜ 0.05 ～ 0.01）,conversely could increase the MDA,NO content,LDH activity and Ca2＋ levels（P＜0.05～0.01）.Each dose of drugs groups could increase the SOD activity and SOD inhibition rate,conversely could reduce the LDH activity,NO content,MDA and Ca2＋ levels when compared with the hydrogen peroxide model group showing the significant differences（P＜0.05～0.01）.CONCLUSION Tongsaimai decoction has revealed the significant protective effects towards the PC12 cells injury,its mechanisms are closely-related to the calcium overload,the oxidative stress and others.

关 键 词：通塞脉 过氧化氢 PC12细胞 氧化应激 

分 类 号：R285.5[医药卫生—中药学]