灯盏花素对缺血再灌注鼠脑海马三磷酸腺苷含量及其活性变化的干预

背景灯盏花素作为蛋白激酶C的抑制剂,具有降低脑缺血再灌注引起的神经细胞死亡作用.目的探讨灯盏花素对缺血再灌注沙土鼠脑海马中三磷酸腺苷的含量,以及对三磷酸腺苷酶活性的影响.设计随机对照区组设计.单位苏州大学附属四院麻醉科,无锡市第四人民医院麻醉科,徐州医学院附属医院麻醉科.材料实验于2002-03/2003-05在江苏省麻醉学重点实验室完成.选取蒙古沙土鼠90只,随机分为9组假手术组、脑缺血组、脑缺血再灌注1 h组、脑缺血再灌注12 h组、脑缺血再灌注24 h组、灯盏花素+脑缺血组、灯盏花素+脑缺血再灌注1 h组、灯盏花素+脑缺血再灌注12 h组、灯盏花素+脑缺血再灌注24 h组,10只/组.灯盏花素注射液(云南省生物制药厂生产,批号990103,每毫升含黄酮4.5 mg).方法除假手术组外,其余各组均建立缺血再灌注模型.灯盏花素各组在缺血前30 min腹腔注射灯盏花素10 mL/kg,其余组同期注射等量生理盐水.每组抽取6只行三磷酸腺苷含量及三磷酸腺苷酶活性测定,其余4只进行海马CA1区神经细胞病理学检查.在缺血期间监测鼓膜温度(反映脑温),并维持鼓膜温度于(37±0.2)℃,以避免温度变化对实验结果的影响.组间比较采用单因素方差分析.主要观察指标①各组实验鼠脑海马三磷酸腺苷含量和三磷酸腺苷酶活性的变化.②各组实验鼠脑海马CA1区神经细胞病理学变化.结果实验纳入蒙古沙土鼠90只,全部进入结果分析.①各组海马三磷酸腺苷含量和三磷酸腺苷酶活性的变化与脑缺血再灌注24 h组比较,灯盏花素1,12,24 h组三磷酸腺苷含量均明显提高[(0.25±0.08),(0.81±0.12),(0.58±0.07),(0.43±0.09)mmol/kg,P＜0.01],Na+-K+-ATP酶活性均明显上升[(1.23±0.43),(5.09±0.36),(3.67±0.37),(2.71±0.42)mmol/(g·h),P＜0.01],Ca2+-ATP酶活性亦明显上升[(0.58±0.35),(2.14±0.41),(1.64±0.39),(1.39±0.40)mmol/(g·h),P＜0.01].②各组海马CA1区神经细胞病理学变化脑缺血后7 d,低倍镜下见脑缺血再灌注1,12,24 h组锥体细胞带变薄、稀疏,甚至消失;而灯盏花素各组锥体细胞带明显比脑缺血再灌注各组宽厚.高倍镜下见脑缺血再灌注1,12,24 h组锥体细胞大部凝固性坏死,核膜完整、核仁清楚的存活锥体细胞数较少;而灯盏花素各组存活锥体细胞明显多于脑缺血再灌注1,12,24 h组.结论灯盏花素能够明显增加脑缺血再灌注后三磷酸腺苷含量,且再灌注初期升高尤为显著,同时三磷酸腺苷酶的活性亦呈相应提高趋势,具有减轻脑缺血再灌注损伤、保护神经细胞的作用.

Effects of erigeron on hippocampus adenosine triphosphate of gerbils with cerebral ischemia reperfusion injury

BACKGROUND: Erigeron, the inhibitor of protein kinase C, functions to decrease the death of neurocytes caused by cerebral ischemia reperfusion.OBJECTIVE: To study the effects of erigeron on hippocampus adenosine triphosphate (ATP) and adenosine triphosphatase (ATPase) of gerbils with cerebral ischemia reperfusion injury.DESIGN: A random controlled experiment.SETTING: Anesthesia Department of the Fourth Hospital of Suzhou University, Anesthesia Department of the Fourth People's Hospital of Wuxi City, and Anesthesia Department of the Affiliated Hospital of Xuzhou Medical College.MATERIALS: The experiment was done in the key laboratory of anesthesiology of Jiangsu Province from March 2002 to May 2003. The 90 Mongolian gerbils were divided into 9 groups: sham operated group, cerebral ischemia group, cerebral ischemia reperfusion 1 hour group, cerebral ischemia reperfusion 12 hours group, cerebral ischemia reperfusion 24 hours group, erigeron + cerebral ischemia group, erigeron + cerebral ischemia reperfusion 1 hour group, erigeron + cerebral ischemia reperfusion 12 hours group, and erigeron + cerebral ischemia reperfusion 24 hours group,with 10 gerbils in each. The erigeron injection with flavone 4.5 g/mL was made by the Biomedicine Factory of Yunnan Province and numbered 990103.METHODS: Apart from the sham operated group, all groups were established the ischemia reperfusion models. The erigeron groups were abdominally injected erigeron 10 mL/kg 30 minutes before the ischemia. Meanwhile, the other groups were injected the normal saline 10 mL/kg. The 6 gerbils of each group were taken blood for testing ATP content and ATPase activity and the rest 4 were only examined the pathological changes of neurocytes of hippocampus CA1 region. During the ischemia, the temperature of drum membrane (reflecting the brain temperature) was monitored and kept at (37±0.2)℃ in order to avoid the influence of temperature on the result. The comparison among groups was shown with the single factor analysis of variance.CA1 region.of ATP content and ATPase activity: Compared with the cerebral ischemia reperfusion 24 hours group, the ATP content of the erigeron 1, 12, 24 hours groups was obviously increased [(0.25 ±0.08), (0.81 ±0.12), (0.58 ±0.07),(0.43±0.09) mmol/kg, P ＜ 0.01], the Na+-K+-ATPase activity was obviously P ＜ 0.01], and the Ca2+-ATPase activity was also obviously promoted days after ischemia, the findings under the low power lens were: the pyramidal belt of the cerebral ischemia reperfusion groups became thinner and loose and even disappeared, but that of the erigeron groups became obviously wider and thicker; the findings under the high power lens were: most of the pyramidal cells of the cerebral ischemia reperfusion groups were in the state of coagulation necrosis, and those survived pyramidal cells with complete nuclear membrane and clear nucleole were very few, but the survived pyramidal cells of the erigeron groups were obviously more than those of the cerebral ischemia reperfusion groups.CONCLUSION: Erigeron increases obviously the ATP content after ischemia reperfusion, especially at the early stage of reperfusion, and at the same time, the ATPase activity is also in an improved tendency, relieving the ischemia reperfusion injury and protecting the neurocytes.