藏红花素预处理对缺血性脑损伤大鼠海马线粒体保护作用的研究  

Study on the protective effect of crocin pretreatment on hippocampal mitochondria in the rats with ischemic brain injury

作　　者：温彬[1] WEN Bin(Baoding First Central Hospital, Baoding 071000, Hebei, China)

机构地区：[1]河北省保定市第一中心医院,河北保定071000

出　　处：《现代中西医结合杂志》2020年第31期3451-3455,3464,共6页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：河北省中医药管理局科研计划项目(2019367)。

摘　　要：目的探讨藏红花素预处理对缺血性脑损伤大鼠海马线粒体的保护作用。方法将120只SD大鼠随机分为6组,每组20只。于术前7 d开始,藏红花素低、中、高剂量组分别腹腔注射10,20,40 mg/kg藏红花素,尼莫地平组腹腔注射1 mg/kg尼莫地平注射液,假手术组、模型组注射等体积生理盐水,均1次/d。干预结束后,除假手术组外,其余组均采用改良线栓法闭塞大脑中动脉的方法复制缺血性脑损伤大鼠模型。造模24 h后,透射电子显微镜观察海马线粒体超微形态结构变化,比色法测定线粒体膜肿胀程度,荧光偏振度法测定线粒体膜流动性,Trinder酶试剂盒测定线粒体膜总磷脂含量,荧光酶标仪测定线粒体膜通透性转换孔(MPTP)开放度和膜电位(△ψm),改良Clark氧电极法测定线粒体呼吸功能和呼吸酶(NADH脱氢酶、琥珀酸脱氢酶和细胞色素C氧化酶)活性,比色法测定线粒体ATP含量,原子化学发光法测定线粒体游离Ca2+浓度。结果藏红花素中、高剂量组和尼莫地平组缺血性脑损伤大鼠海马线粒体超微形态结构病变明显轻于模型组(P均<0.05),线粒体膜肿胀度明显轻于模型组(P均<0.05),膜流动性、膜磷脂含量、△ψm、线粒体呼吸功能和呼吸酶活性、ATP含量均明显高于模型组(P均<0.05),MPTP开放度、游离Ca2+浓度均明显低于模型组(P均<0.05)。结论藏红花素预处理对缺血性脑损伤大鼠海马线粒体结构和功能具有保护作用。Objective It is to investigate the protective effect of crocin pretreatment on hippocampal mitochondria in the rats with ischemic brain injury.Methods 120 SD rats were randomly divided into 6 groups with 20 rats in each group.From the 7th day before operation,the crocin low,medium and high dose groups were injected intraperitoneally with 10,20,and 40 mg/kg crocin,respectively,the nimodipine group was injected intraperitoneally with 1 mg/kg nimodipine injection,and the sham operation group and model group were injected with the same volume of normal saline,all once per day.After the intervention,except for the sham operation group,the rest groups used the modified suture method to occlude the middle cerebral artery to replicate the ischemic brain injury rat models.In 24 h after modeling,transmission electron microscopy was used to observe the ultrastructural changes of hippocampal mitochondria,colorimetric method was used to measure the swelling degree of mitochondrial membrane,fluorescence polarization method was used to measure the fluidity of mitochondrial membrane,Trinder enzyme kit was used to measure total phospholipid content of mitochondrial membrane,fluorescence microplate reader was used to measure mitochondrial mitochondrial permeability transition pore(MPTP)openness and membrane potential(△ψm),modified Clark oxygen electrode method was used to measure mitochondrial respiratory function and respiratory enzymes(NADH dehydrogenase,succinate dehydrogenase and cytochrome C oxidase)activity,colorimetric method was used to determine mitochondrial ATP content,and atomic chemiluminescence method was used to determine the content of mitochondrial free Ca2+.Results The ultrastructural changes of hippocampal mitochondria in rats with ischemic brain injury in the middle and high doses of crocin and nimodipine group were significantly lighter than those in the model group(all P<0.05),and the swelling of the mitochondrial membrane was significantly less than those in the model group(all P<0.05),membrane fluidity,m

关 键 词：藏红花素 缺血性脑损伤 海马 线粒体 膜电位 

分 类 号：R-332[医药卫生]