灯盏细辛胶囊对慢性脑缺血大鼠及缺氧损伤PC12细胞的保护作用  

Dengzhan Xixin(灯盏细辛)Capsules Protect Rats from Chronic Cerebral Ischemia-Induced Brain Injury and PC12 Cells from Hypoxia Injury

作　　者：付英豪 吴秋宇 李朝阳 米佳佳 鄢宇梅 盛艳梅[1] FU Yinghao;WU Qiuyu;LI Zhaoyang;MI Jiajia;YAN Yumei;SHENG Yanmei(School of Pharmacy,Chengdu Medical College,Chengdu 610500)

机构地区：[1]成都医学院药学院,成都610500

出　　处：《中药药理与临床》2024年第1期23-29,共7页Pharmacology and Clinics of Chinese Materia Medica

基　　金：四川生大学生创新创业项目(S202113705092);四川省科技厅重点研发项目(NO:2020YFS0325)。

摘　　要：目的:分别考察灯盏细辛胶囊对慢性脑缺血大鼠的影响及氯化钴诱导的PC12细胞缺氧损伤的影响,探究其对慢性缺血性脑神经损伤的保护作用。方法:(1)体内实验:双侧颈总动脉永久结扎法制备大鼠慢性脑缺血模型,分别设假手术对照组、模型对照组、灯盏细辛120 mg/kg组、尼莫地平16 mg/kg组,给药8 w后,开展水迷宫认知功能评价;检测血清MDA、SOD、GSH-px含量或活力,同时取皮层组织行HE染色及Western blot法检测AKT、p-AKT、Cleaved-Caspase3蛋白表达。(2)体外实验:确定灯盏细辛胶囊对PC12细胞的IC50,建立氯化钴诱导的PC12细胞缺氧损伤模型,MTT法评价药物对模型细胞增殖的影响,并采用AnnexinVFITC/PI双染法评价药物对细胞凋亡的影响。结果:(1)水迷宫试验:与假手术对照组比较,模型对照组逃避潜伏期显著延长,穿越平台次数显著降低(P<0.01);血清MDA含量显著升高,SOD、GSH-px活力显著降低,p-AKT/AKT比值明显降低(P<0.05),Cleaved-Caspase3蛋白表达显著上调(P<0.01);模型对照组大鼠脑皮层细胞形态异常,见较多神经元细胞固缩深染,胞质与胞核分界不清;与模型对照组比较,灯盏细辛120 mg/kg组能明显缩短大鼠定位航行试验中的逃避潜伏期、寻找平台的运动距离,增加空间探索试验中穿越平台的次数(P<0.01);血清MDA含量显著降低,SOD、GSH-px活力明显升高(P<0.05或P<0.01),p-AKT/AKT比值明显增加,Cleaved-Caspase3蛋白表达显著下调(P<0.01),大鼠脑组织病理损伤明显减轻。(2)体外实验:与正常对照组比,模型对照组细胞凋亡率显著升高(P<0.01);灯盏细辛IC50值为0.84 mg/mL;与模型对照组比较,灯盏细辛在一定浓度内能促进氯化钴诱导的PC12细胞增殖,并降低其凋亡率(P<0.05或P<0.01)。结论:灯盏细辛胶囊可以对抗慢性脑缺血大鼠的氧化损伤以改善其认知功能,可能通过激活AKT/Caspase3信号通路对抗缺血性神经细胞凋亡产生脑神经保护作�Objective:To investigate the effects of Dengzhan Xixin(灯盏细辛)Capsules(DZXXC)on chronic cerebral ischemia in rats and cobalt chloride-induced hypoxia injury in PC12 cells,and to explore the neuroprotective effect of DZXXC on chronic ischemic injury.Methods:(1)Animal experiment:The rat model of chronic cerebral ischemia was prepared by permanent ligation of bilateral common carotid arteries.The model,sham operation,DZXXC(120 mg/kg),and nimodipine(16 mg/kg)groups were set up respectively.After 8 weeks of administration,the Morris water maze test was carried out to examine the cognitive function of rats,and the levels of malondialdehyde(MDA),superoxide dismutase(SOD),and glutathione peroxidase(GSH-px)in the serum were determined.Meanwhile,the cortical tissue was stained with hematoxylin-eosin and examined for the expression of protein kinase B(AKT),phosphorylated AKT(p-AKT),and cleaved caspase 3 by Western blotting.(2)Cell experiment:The half maximal inhibitory concentration(IC50)of DZXXC on PC12 cells was measured,and the PC12 cell model of hypoxia injury was established with cobalt chloride.The proliferation of the model cells with and without drug treatment was evaluated by the methyl thiazolyl tetrazolium(MTT)method.The apoptosis of the cells was evaluated by the annexin VFITC/PI double staining method.Results:(1)Animal experiment:Compared with the sham operation group,the model group showed increased latency and reduced number of crossing the platform in the Morris water maze test(P<0.01),elevated MDA level and lowered SOD and GSH-px levels in the serum,decreased p-AKT/AKT ratio(P<0.05),and up-regulated expression of cleaved Caspase 3(P<0.01).Moreover,the model group presented abnormal cell morphology,karyopyknosis of neurons,and obscure boundaries between cytoplasm and nucleus.Compared with the model group,DZXXC shortened the latency and the movement distance of finding platforms in the navigation test,and increased the number of crossing the platform in the spatial probe test(P<0.01).Furthermore,DZXXC l

关 键 词：灯盏细辛胶囊 慢性脑缺血 PC12细胞 缺氧损伤 蛋白激酶B 半胱氨酸蛋白酶3 

分 类 号：R285.5[医药卫生—中药学]