外源性一氧化碳后处理对失血性休克大鼠平衡能力的改善:与细胞外调节蛋白激酶1/2的关系  

Post-conditioning with exogenous carbon monoxide improves the ability of balance in rats exposed to hemorrhagic shock and resuscitation:the roles of extracellular regulated protein kinase 1/2

作　　者：张立民[1] 张冬雪[2] 孙文波[1] 李睿[1] ZHANG Limin ZHANG Dongxue SUN Wenbo LI Rui(Department of Anesthesiology Department of Gerontology, Cangzhou Central Hospital, Cangzhou 061001, China)

机构地区：[1]沧州市中心医院麻醉科,河北沧州061000 [2]沧州市中心医院老年内科,河北沧州061000

出　　处：《麻醉安全与质控》2017年第4期185-190,共6页Perioperative Safety and Quality Assurance

基　　金：国家自然科学基金面上项目(81473285)

摘　　要：目的探讨外源性一氧化碳(CO)后处理对失血性休克后大鼠平衡能力的改善以及对小脑皮质内细胞外调节蛋白激酶1/2(ERK1/2)的影响。方法雄性SD大鼠60只,体质量(350~400)g,根据随机数字表法,将大鼠分为5组(n=12),分别为对照组(Sham组)、失血性休克组(H组)、失血性休克+CO后处理组(HC组)、PD98059+失血性休克+CO后处理组(PHC组)、PD98059+失血性休克组(PH)。失血性休克通过股静脉放血使平均动脉压为(30±5)mm Hg维持60 min建立模型,再将收集的血液回输至大鼠体内达到初始血压水平作为复苏,必要时输注生理盐水;将大鼠置于含有10 m L/L CO的玻璃箱内3 h作为外源性CO后处理;ERK1/2阻断处理为放血前1 h向脑室内注射30μL的ERK1/2抑制剂PD98059(30μmol/L);对照组只进行股静脉、股动脉穿刺置管和脑室内等量生理盐水注射。采用气相分析法测定小脑皮质中CO含量,采用杠杆秤和倾斜测试评价大鼠的平衡能力,采用HE染色法测定嗜酸性浦肯野细胞数量,采用TUNEL法测定神经元凋亡率,采用免疫蛋白印迹法测定磷酸化ERK1/2、总磷酸化ERK1/2、Bcl-2和Bax蛋白表达。结果与Sham组比较,H组小脑皮质内CO含量增加,神经细胞凋亡率和嗜酸性浦肯野细胞数增加,杠杆秤和倾斜测试的合格率下降,磷酸化ERK1/2水平升高,Bcl-2/Bax比值下降(P<0.05);与H组比较,HC组小脑皮质内CO含量增加,神经细胞凋亡率和嗜酸性浦肯野细胞数减少,杠杆秤和倾斜测试的合格率升高,磷酸化ERK1/2水平升高,Bcl-2/Bax比值升高(P<0.05);与HC组比较,PHC组神经元凋亡率和嗜酸性浦肯野细胞数增加,杠杆秤和倾斜测试的合格率下降,磷酸化ERK1/2水平降低,Bcl-2/Bax比值下降(P<0.05)。结论外源性CO改善失血性休克大鼠平衡能力的机制,与增加小脑皮质内神经细胞ERK1/2磷酸化水平,降低小脑皮质神经细胞凋亡密切相关。Objective To evaluate whether post-conditioning with exogenous carbon monoxide （CO） improves ability of balance in rats exposed to hemorrhagic shock and resuscitation via roles of extraeellular regulated protein kinase 1/2 signal pathway. Methods In accord- ance with the random number table, 60 male SD rats with body weight between 350 to 400 g were divided into 5 groups（n = 12 each）, including the Sham group, the group of hemorrhagic shock and the resuscitation （ H groups） , the group of hemorrhagic shock and resusci- tation plus CO post-conditioning （ HC group）, the group of hemorrhagic shock and resuscitation plus PD98059 and CO post-conditioning （PHC group）, and the group of hemorrhagic shock and resuscitation plus PD98059 treatment （PH group）. The rats in H, PHC and PH groups were subjected to hemorrhage by bleeding from femoral vein to achieve mean arterial pressure of 30 mmHg for 60 min followed by resuscitation with shed blood and saline as needed to restore blood pressure. CO post-conditioning was established by 1% CO exposure for 3 h after resuscitation. In PHC and PH groups PD98059 was injected into cerebral ventricle to inhibit phosphorylation of ERK1/2. The con- tent of CO was assessed by gas chromatograph assay; the balance ability was assessed by beam balance and inclined plane test; the acido- philia purkinje cells were assessed using HE staining; the effects of cerebellar apoptosis were assessed by TUNEL assays; the expression of phosphorylational, total phosphorylational ERK1/2, Bcl-2 and Bax was detected by Western blot. Results Compared with the corre- sponding findings in Sham group, there were significant increases in CO content, acidophilia purkinje ceils, neuronal apoptosis, and ERK1/2 phosphorylation ; and decreases in beam balance and inclined plane score, ratio of Bcl-2/Bax in H group （ P 〈 0.05）. Compared with corresponding findings in H group, there were significant decreases in acidophilia purkinje cells, neuronal apoptosis and increases in CO content, E

关 键 词：一氧化碳 缺氧 小脑皮质 细胞凋亡 后处理 

分 类 号：R605.971[医药卫生—急诊医学]