人参皂苷Rg1抗新生鼠缺氧缺血性脑损伤后神经元凋亡的研究  

ANTI-APOPTOTIC EFFECT OF GINSENOSIDE Rg1 ON NEURON AFTER NEONATAL HYPOXIA ISCHEMIA BRAIN DAMAGE

作　　者：王德健[1,2] 黄源芳[1] 李巧云[2] 徐世军[2] 刘小康[1] 

机构地区：[1]四川大学华西基础医学与法医学院药理教研室,成都610041 [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》2010年第9期1107-1112,共6页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：成都中医药大学科技发展基金(ZRYB200939)~~

摘　　要：目的人参皂苷Rg1可增强神经元对缺氧缺血应激的耐受能力,在缺氧缺血性脑损伤(hypoxia ischemia brain damage,HIBD)中发挥一定的抗凋亡作用。观察人参皂苷Rg1对新生鼠HIBD后神经元凋亡及神经功能恢复的影响,并探讨其可能机制。方法 10天龄SPF级SD大鼠54只,体重16～22g,随机分为假手术对照组(假手术组,n=6)、HIBD模型组(模型组,n=24)和人参皂苷Rg1组(Rg1组,n=24)。模型组及Rg1组大鼠采用结扎右侧颈总动脉并低氧通气制备HIBD模型;假手术组仅分离右侧颈总动脉,不结扎,不进行低氧通气。Rg1组于术后即刻腹腔内注射0.1mL含人参皂苷Rg1(40mg/kg)的生理盐水,此后每隔24h按相同剂量注射人参皂苷Rg1;模型组和假手术组相同时间点腹腔内注射0.1mL生理盐水。术后观察大鼠一般情况,于术后4、8、24、72h采用Longa评分法行神经行为学评价后,处死大鼠取右侧脑组织,采用Western blot及免疫组织化学染色检测缺氧诱导因子1α(hypoxia inducible factor1α,HIF-1α)和活化的半胱天冬氨酸酶3(cleaved caspase3,CC3)蛋白表达;TUNEL法检测原位神经元凋亡情况。结果术后大鼠均存活至实验完成。与假手术组比较,模型组和Rg1组大鼠均出现不同程度的神经行为学异常;两组Longa评分与假手术组比较,差异均有统计学意义(P<0.05);术后72h时Rg1组与模型组比较,差异有统计学意义(P<0.05)。Western blot检测显示,各组各时间点均有HIF-1α、CC3蛋白表达;模型组及Rg1组各时间点HIF-1α蛋白表达均较假手术组有明显增加(P<0.05),Rg1组均较同一时间点模型组有明显上调(P<0.05)。模型组各时间点CC3蛋白表达均较假手术组明显增加(P<0.05),Rg1组仅术后4h与假手术组比较,差异有统计学意义(P<0.05);Rg1组各时间点均较模型组明显下调(P<0.05)。免疫组织化学染色可见HIF-1α蛋白、CC3蛋白定位主要集中在胞核及胞浆,各组各时间点蛋白表达强度与Western blot观察�Objective Ginsenoside Rg1 could increase the tolerance of neural hypoxia and ischemia under stress, and play an anti-apoptotic effect in hypoxia ischemia brain damage （HIBD）. To investigate the effects of ginsenoside Rg1 on neural apoptosis and recovery of neurological function in neonatal rats with HIBD, and to explore the possible mechanism. Methods Fifty-four 10-day-old SD rats （weighing 16-22 g） were randomly allocated into sham-operation group （Sham group, n=6）, HIBD model group （HIBD group, n=24）, and ginsenoside Rg1 treatment group （Rg1 group, n=24）. SD rats in HIBD group and Rg1 group were made the models of HIBD by ligation of the right common carotid artery （CCA） and subsequently hypoxic ventilation （8%O2 plus 92%N2） for 2.5 hours; and in Sham group, the right CCA was only exposed without ligation of CCA and hypoxic ventilation. Intraperitoneal injection of 0.1 mL normal saline （NS） containing 40 mg/kg Rg1 was given immediately after operation in Rg1 group, intraperitoneal injection of 0.1 mL pure NS was given in both HIBD group and Sham group and was repeated every 24 hours. The general state of SD rats was monitored after operation, and Longa scores were recorded to evaluate the neurological function at 4, 8, 24, and 72 hours after HIBD. Western blot and immunohistochemistry staining were used to detect protein expressions of both hypoxia inducible factor 1α （HIF-1α） and cleaved caspase 3 （CC3）. TUNEL staining was used to evaluate neural apoptosis in situ. Results All rats survived to the end of the experiment. Neurological dysfunction was observed in both HIBD group and Rg1 group, showing significant difference in Longa score when compared with that in Sham group （P〈0.05）. There was significant difference in Longa score between Rg1 group and HIBD group at 72 hours after HIBD （P〈0.05）. Western blot showed that the protein expressions of both HIF-1α and CC3 were observed at every time point in every group. The expressions of HIF-1α protein in HIBD

关 键 词：缺氧缺血性脑损伤 神经元凋亡 人参皂苷RG1 缺氧诱导因子1Α 活化的半胱天冬氨酸酶3 大鼠 

分 类 号：R285[医药卫生—中药学]