胞外信号相关蛋白激酶1/2信号通路与人参皂苷Rg1抗新生鼠缺氧缺血性脑损伤后神经元凋亡  

ROLE OF EXTRACELLULAR SIGNAL-RELATED PROTEIN KINASE 1/2 PATHWAY IN GINSENOSIDE Rg1 MEDIATED ANTI-APOPTOTIC EFFECT ON NEURON AFTER HYPOXIA ISCHEMIA BRAIN DAMAGE IN NEONATAL RATS

作　　者：唐彬秩[1] 王德健[2] 吴青[1] 李茂军[1] 阳倩[1] 陈昌辉[1] 

机构地区：[1]四川省医学科学院.四川省人民医院儿科,成都610072 [2]成都中医药大学药学院药理学教研室

出　　处：《中国修复重建外科杂志》2016年第8期1011-1018,共8页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：四川省人民医院博士基金资助项目(30305030580);成都中医药大学科技发展基金资助项目(030029052)~~

摘　　要：目的探讨人参皂苷Rg 1在新生鼠缺氧缺血性脑损伤（hypoxia ischemia brain damage,HIBD）中的抗凋亡作用,分析可能的信号通路机制。方法 10日龄SPF级SD大鼠48只,体质量17~21 g,随机分为4组（n=12）,假手术组、模型组（HI组）、模型＋人参皂甙Rg 1组（HI＋Rg 1组）、模型＋人参皂甙Rg 1＋U 0126干预组（HI＋Rg 1＋U 0126组）。HI组、HI＋Rg 1组及HI＋Rg 1＋U 0126组大鼠采用结扎单侧颈总动脉并低氧通气方法制备HIBD模型;假手术组仅分离右侧颈总动脉。HI＋Rg 1＋U 0126组于造模前1 h右侧脑室注射5μL含U 0126（25μg/kg）的PBS,其余3组同法注射5μL PBS。HI＋Rg 1组及HI＋Rg 1＋U 0126组于术后即刻腹腔内注射0.1 m L含Rg 1（40 mg/kg）的生理盐水;HI组和假手术组注射0.1 m L生理盐水。术后4、24 h处死各组大鼠,取右侧半球皮层和海马脑组织,采用Western blot及免疫组织化学染色检测胞外信号相关蛋白激酶1/2（extracellular signalrelated protein kinase 1/2,Erk 1/2）及磷酸化Erk 1/2（phospho-Erk 1/2,p-Erk 1/2）、缺氧诱导因子1α（hypoxia inducible factor 1α,HIF-1α）和活化的半胱天冬氨酸酶3（cleaved Caspase-3,CC 3）蛋白表达;TUNEL法检测原位神经元凋亡情况。结果 Western blot检测示,各时间点各组均有Erk 1/2、p-Erk 1/2、HIF-1α、CC 3蛋白表达;术后4、24 h,HI组HIF-1α、CC 3蛋白表达均较假手术组明显增加（P〈0.05）;术后4 h,HI组p-Erk 1/2蛋白表达较假手术组明显增加（P〈0.05）;术后4、24 h,HI＋Rg 1组p-Erk 1/2及HIF-1α蛋白表达较HI组明显上调（P〈0.05）,CC 3蛋白表达则明显下调（P〈0.05）;术后4、24 h,HI＋Rg 1＋U 0126组p-Erk 1/2及HIF-1α蛋白表达较HI＋Rg 1组明显下调（P〈0.05）,CC 3蛋白表达则明显上调（P〈0.05）。各时间点各组间Erk 1/2蛋白表达比较,差异均无统计学意义（P〉0.05）。免疫组织化学染色可见HIF-1α蛋白、CC 3蛋白定位主要集中在细胞核及细胞质Objective To investigate the anti-apoptotic effect of ginsenoside Rg 1 in neonatal rats with hypoxia ischemia brain damage（HIBD）, and to explore the possible signaling pathway involved in anti-apoptosis. Methods Forty-eight 10-day-old Sprague Dawley（SD） rats（weighing 17- 21 g, male or female） were randomly allocated into 4 groups（ 12 rats in each group）： sham-operation group（sham group）, HIBD group（HI group）, HIBD＋ginsenoside Rg 1 group（HI＋Rg 1 group）, and HIBD＋ginsenoside Rg 1 ＋U 0126 group（HI＋Rg 1 ＋U 0126 group）. SD rats in HI group, HI＋Rg 1 group, and HI＋Rg 1 ＋U 0126 group underwent ligation of the right common carotid artery（CCA） and hypoxic ventilation（ 8 %O 2 ＋ 92 %N 2） for 2. 5 hours to prepare the HIBD model, and rats in sham group underwent only separation of the right CCA. SD rats in HI＋Rg 1 ＋U 0126 group received intraventricular injection of 5 μL phosphate buffer saline（PBS） containing U 0126（ 25 μg/kg） at 1 hour before HIBD, and rats in the other three groups received intraventricular injection of PBS at the same time. The rats in HI＋Rg 1 group and HI＋Rg 1 ＋U 0126 group received intraperitoneal injection of 0. 1 m L normal saline（NS） containing Rg 1（ 40 mg/kg） at immediate after HIBD, while rats in HI group and sham group received intraperitoneal injection of 0. 1 m L NS at immediate after HIBD. At 4 and 24 hours after HIBD, the right hemisphere and hippocampus were collected to detect the protein expression and distribution of extracellular signal-related protein kinase 1 / 2（Erk 1 / 2）, phospho-Erk 1 / 2（p-Erk 1 / 2）, hypoxia inducible factor 1 α（HIF- 1 α）, and cleaved Caspase- 3（CC 3） by Western blot and immunohistochemistry staining. TUNEL staining was used to evaluate neural apoptosis in situ. Results Western blot results showed that there were expressions of Erk 1 / 2, p-ERK 1 / 2, HIF-1 α, and CC 3 proteins at 4 and 24 hours after HIBD in each group. The expressions of HIF- 1 α and CC

关 键 词：缺氧缺血性脑损伤 神经元凋亡 人参皂苷RG1 胞外信号相关蛋白激酶1/2 大鼠 

分 类 号：R285.5[医药卫生—中药学]