生地注射液对凝血酶损伤后神经元保护的实验研究  

An experimental study of protective effects of Shengdi (生地) injection on neuron injured by thrombin

作　　者：刘爱华[1] 俞晓飞[1] 汪洋[2] 魏江磊[1] 

机构地区：[1]上海中医药大学附属曙光医院,上海 200021 [2]上海复旦大学医学院解剖组胚教研室,上海 200032

出　　处：《中国中西医结合急救杂志》2008年第3期155-158,共4页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：上海市教委课题资助项目(05cz19)

摘　　要：目的观察凝血酶对神经元的作用,并筛选凝血酶损伤神经元的最小剂量,探讨生地注射液对神经元损伤的保护作用。方法培养胎鼠大脑皮质神经元细胞。将接种于96孔板的神经元培养7d后分为6组,每组12个孔,依次加入0(对照组)、25、50、75、100和150kU/L凝血酶,观察凝血酶对神经元的损伤作用。将接种于96孔板的神经元培养7d后分为12组,每组8个孔,设正常对照组,其余11组分别加入75kU/L凝血酶作用24h后,依次加入体积分数为1%、2%、3%、3.5%、4%、4.5%、5%、5.5%、6%、8%、10%的生地注射液。观察各组神经元病理形态学改变;以四甲基偶氮唑盐(MTT)比色法和乳酸脱氢酶(LDH)释放量来反映神经元数量和活性。结果与对照组比较,25kU/L凝血酶组LDH显著降低,差异有统计学意义(P<0.05),其他组LDH值随凝血酶剂量增加呈增高趋势,且75、100和150kU/L凝血酶组与对照组比较差异有统计学意义(P均<0.01)。与对照组比较,除25kU/L凝血酶组MTT值升高外,其他组MTT值均显著下降,且与对照组比较差异均有统计学意义(P均<0.01);综合LDH与MTT值的结果,筛选出凝血酶损伤神经元的最小剂量为75kU/L。在生地注射液干预实验中,与正常对照组比较,生地注射液各浓度组LDH值均降低,MTT值均升高,除10%组MTT值外,其余各值间差异有统计学意义(P<0.05或P<0.01)。光镜和电镜下观察正常对照组和生地注射液各浓度组神经元细胞形态均显示正常;凝血酶各浓度组细胞则有不同程度损伤。结论生地注射液能够提高凝血酶损伤后神经元存活率、抑制LDH释放,从而对神经元起到保护作用。Objective To investigate the effect of thrombin on neuron, select the least dosage of thrombin that may injure neuron and approach the protective effect of Shengdi （生地, a herb of traditional Chinese medicine）on neuronal injury. Methods The fetus rat neurons of cerebral cortex were cultured, the cultured cells were inoculated in 96 well-plates for 7 days, then they were divided into 6 groups, each group being in 12 wells. Afterwards, the different concentrations of thrombin were added in each well in turn, that was 0 （normal group）, 25, 50, 75, 100 and 150 kU/L to investigate the thrombin injuring effect on the neuron. The neurons which were cultured in 96 well-plates for 7 days were divided into 12 groups, and there were 8 wells in each group. Except for the normal group, in the other groups, 75 kU/L thrombin was added, after its action took for 24 hours, then Shengdi injection in different volume fractions which were 1%, 2%, 3%, 3.5%, 4%,4.5%, 5%, 5.5%, 6%, 8%, 10% consecutively put into the well plates. After that, the morphological changes of neuron were observed. At the same time, the indexes of survival rate [methyl thiazdyl tetrazolium （MTT）] and injury [lactic dehydrogenase （LDH）] were tested to show the number and activity of the neuron. Results Compared with the normal group, the value of LDH in 25 kU/L group was decreased and showed statistical significance （P〈0.05）, while in the other groups, it was suggested that the increase of LDH tended to follow the increase of dosage of thrombin, and there were statistical significances in 75, 100 and 150 kU/L thrombin groups （all P〈0.01）. On the other hand, compared with the normal group, except for the 25 kU/L group in which the value of MTT was increased, in all the other groups it was decreased significantly （all P〈0.01）. According to the results of LDH and MTT, the selected least dosage that might injure the neuron was 75 kU/L thrombin. In the experiment of intervention by Shengdi injection, compared with the normal grou

关 键 词：生地注射液 凝血酶 神经元 乳酸脱氢酶 

分 类 号：R285.5[医药卫生—中药学] Q813.11[医药卫生—中医学]