参附注射液对缺氧缺血性脑损伤新生大鼠脑保护机制研究  

Neuroprotective effects of Shenfu injection on hypoxic-ischemic brain damage in neonatal rats

作　　者：刘文强[1] 王军[1] 徐艳[1] 韩爱民[1] 杨倩倩[1] 

机构地区：[1]徐州医学院附属医院新生儿科,221002

出　　处：《中国神经免疫学和神经病学杂志》2015年第3期200-205,共6页Chinese Journal of Neuroimmunology and Neurology

基　　金：徐州市科技发展基金计划项目(XF10C066)

摘　　要：目的探讨参附注射液对缺氧缺血性脑损伤(HIBD)新生大鼠皮质区钙离子相关蛋白钙网蛋白(CRT)的表达、细胞内游离Ca2+浓度以及神经元凋亡的影响,探讨其可能的脑保护机制。方法将7日龄新生SD大鼠随机分为对照组、缺氧缺血组和参附干预组,各组进一步分为缺氧缺血后3、6、12、24、72h5个亚组,每组均10只。参照Rice等法制备HIBD模型。参附干预组于造模成功后立即腹腔注射参附注射液10mL/(kg·d),连用3d。缺氧缺血组和对照组注射等量生理盐水。HE染色观察大鼠病变侧脑组织病理变化;RT-PCR及Western-blot法检测各组CRT的表达变化;荧光显微镜法检测脑细胞中游离Ca2+浓度;流式细胞术检测神经元凋亡率。结果缺氧缺血组和参附干预组各时间点CRT mRNA及蛋白表达量均较对照组升高(P<0.05),且参附干预组较缺氧缺血组升高(P<0.05);缺氧缺血组各时间点细胞内游离Ca2+浓度均较对照组明显升高(P<0.05),参附干预组细胞内游离Ca2+浓度较缺氧缺血组降低(P<0.05),但仍高于对照组(P<0.05);参附干预组各时间点的神经元凋亡率均明显低于缺氧缺血组(P<0.05),但较对照组仍升高(P<0.05)。结论参附注射液可能通过上调CRT的表达,降低细胞内钙超载来减轻新生大鼠缺氧缺血性脑损伤。Objective To investigate the expression of calreticulin （CRT）, the change of intracellular free calcium and neuronal apoptosis in cerebral cortex of neonatal rats with hypoxic-ischemic brain damage （HIBD）, and the intervention effects of Shenfu injection. Methods Sprague-Dawley （SD） rats aged 7 days were randomly assigned to three groups： control group, hypoxic ischemic group and Shenfu treated group. Rat models of HIBD were established according to Rice＇s method. Based on the observing time points, each group （n= 50） was subdivided into 5 subgroups sacrificed at 3, 6, 12, 24 and 72 hrs. Rats in the control group were not performed ligation of the right common carotid artery and not underwent hypoxia but only performed the separation of right common carotid artery. Shenfu injection was administered by intraperitoneal injection right after H/I insult and then once daily at a dosage of 10 mL/kg for 3 days in the rats of Shenfu treated group. Saline was also administered into the rats of hypoxic ischemic group and control group by the same methods and the same dosage. The pathological changes of right cerebral cortex were observed under light microscope. The expression of CRT in the cerebral cortex was detected by RT-PCR and western blot. The cells of right cerebral cortex were isolated and incubated with Fura-2/AM and observed under fluorescent microscope. The free calcium concentrations were determined as the ratio of F340/F180 via image analysis system. The apoptosis rate was measured by flow cytometry. Results Compared with the control group, the expressions of CRT in hypoxic- ischemic group and Shenfu treated group were obviously up regulated （P〈0.05）, and the expressions of CRT in Shenfu-treated group were notably higher than those in hypoxic-ischemie group （P〈0.05）. The concentrations of intracellular free calcium in Shenfu-treated group were significantly decreased compared with hypoxic-ischemic group （P〈0.05）, but was also higher than those in control group （P〈0.05�

关 键 词：参附注射液 缺氧缺血性脑损伤 钙网蛋白 凋亡 

分 类 号：R743.3[医药卫生—神经病学与精神病学]