石甘散联合丙戊酸钠对癫痫模型幼鼠脑损伤保护作用和认知功能的影响

Protective effect of Shigan powder combined with sodium valproate on brain injury and influence on cognitive function in young rats with epilepsy

目的探讨石甘散联合丙戊酸钠对癫痫模型幼鼠脑损伤保护作用及对其认知功能的影响。方法采用随机数表法将60只健康Wistar幼鼠分为对照组、模型组、丙戊酸钠组、石甘散组和联合组共五组,每组各12只。给予对照组幼鼠连续28天腹腔注射1 ml 0.9%氯化钠注射液,其余四组幼鼠连续28天腹腔注射1 ml 2 mg/ml戊四氮溶液。建模成功后,给予对照组和模型组幼鼠每日2次0.9%氯化钠注射液灌胃,给予丙戊酸钠组幼鼠每日1次200 mg/kg丙戊酸钠灌胃,给予石甘散组幼鼠每日1次1 g/kg石甘散溶液灌胃,给予联合组幼鼠每日1次200 mg/kg丙戊酸钠灌胃和1次1 g/kg石甘散溶液灌胃,所有幼鼠均连续灌胃治疗28天。治疗结束后进行Morris水迷宫测试,随后处死幼鼠并提取海马组织血液,分别采用TUNEL法和JC-1法检测海马组织神经元凋亡情况和神经元线粒体跨膜电位(Δψm),采用酶联免疫吸附法检测血清神经元特异性烯醇化酶(NSE)。结果(1)对照组幼鼠1-5天的逃避潜伏期显著短于其余四组(P<0.01);丙戊酸钠组、石甘散组和联合组幼鼠1-5天的逃避潜伏期均显著短于模型组;联合组幼鼠1-5天的逃避潜伏期显著短于丙戊酸钠组和石甘散组(P<0.01)。(2)五组幼鼠平台穿越次数比较有显著差异,对照组幼鼠穿越次数最多,其次为联合组,模型组幼鼠穿越次数最少(P<0.01)。(3)五组幼鼠海马组织神经元细胞凋亡比例比较有显著差异,对照组比例最低,其次为联合组,模型组最高(P<0.01)。(4)五组幼鼠海马组织神经元Δψm比较有显著差异,对照组Δψm最高,其次为联合组,模型组最低(P<0.01)。(5)五组幼鼠血清NSE水平比较有显著差异,对照组NSE水平最低,其次为联合组,模型组最高(P<0.01)。(6)丙戊酸钠组和石甘散组幼鼠1-5天的逃避潜伏期、平台穿越次数、海马组织神经元细胞凋亡比例、海马组织神经元Δψm以及血清NSE比较无显著差异(P>0.05)。结论石甘散联合丙戊酸钠能够有效改善癫痫幼鼠认知功能,阻断线粒体异常电位保护线粒体功能,抑制海马组织神经元凋亡,减轻脑损伤程度。

Objective To investigate the protective effect of Shigan powder combined with sodium valproate on brain injury in young rats with epilepsy and its influence on cognitive function of the rats.Methods According to random number table method,60 healthy youngWistar rats were randomly divided into control group,model group,sodium valproate group,Shigan powder group and combination group,with 12 young rats in each group.The young rats in the control group were given intraperitoneal injection of 0.9%sodium chloride(1 ml) for 28 consecutive days,and the other four groups were given intraperitoneal injection of 2 mg/ml(1 ml)pentylenetetrazol solution for 28 consecutive days.After successful modeling,young rats in the control group and model group were given 0.9%sodium chloride injection twice a day by gavage,young rats in the sodium valproate group were given 200 mg/kg sodium valproate once a day by gavage,young rats in the Shigan powder group were given 1 g/kg Shigan powder solution once a day by gavage,young rats in the combined group were given 200 mg/kg sodium valproate once a day by gavage and 1 g/kg Shigan powder solution once a day by gavage,and all young rats were given continuous gavage treatment for 28 days.Morris water maze test was carried out after the treatment,and then the young rats were killed and the hippocampus blood was extracted.TUNEL method and JC-1 method were used to detect the neuronal apoptosis and neuronal mitochondrial transmembrane potential in the hippocampus(Δψm).Serum neuron specific enolase(NSE)was detected by enzyme-linked immunosorbent assay.Results The escape latency of the control group in 1-5 days was significantly shorter than those of the other four groups(P<0.01).The escape latency in 1~5 days of the sodium valproate group and the Shigan powder group and the combination group were significantly shorter than those in the model group.The escape latency of young rats in the combination group was significantly shorter than that in the sodium valproate group and the Shigan powder group(P<0.01).There was a significant difference in the crossing times among the five groups of young rats.The control group had the most crossing times,followed by the combination group,and the model group had the least crossing times(P<0.01).There was a significant difference in the percentage of neuronal apoptosis in hippocampus among the 5 groups.The percentage in the control group was the lowest,followed by the combination group,and the model group was the highest(P<0.01).There was a significant difference inΔψm of neurons in hippocampus of 5 groups of young rats,Δψm in the control group was the highest,followed by the combination group,and the model group was the lowest(P<0.01).There was a significant difference in the serum NSE level among the 5 groups.The NSE level in the control group was the lowest,followed by the combination group,and the model group was the highest(P<0.01).Escape latency,platform crossing times,apoptosis ratio of neurons in hippocampus,ΔψM in neurons in hippocampus and serum NSEof young rats in sodium valproate group and Shigan powder group in 1-5 days had no significant difference(P>0.05).Conclusions Shigan powder combined with sodium valproate could effectively improve cognitive function of epileptic young rats,block mitochondrial abnormal potential to protect mitochondrial function,inhibit neuronal apoptosis in hippocampus,and reduce brain damage.