丹参酮ⅡA对新生大鼠缺氧缺血性脑损伤磷酸化NMDA受体1表达及细胞内游离钙浓度的影响.pdf

中南大学学报(医学版)
2010,35(9)http://xbyx.xysm.nel
940
J Cent South Uniy (Med Sci)
丹参酮IA对新生大鼠缺氧缺血性脑损伤
磷酸化NMDA受体1表达及细胞内游离钙浓度的影响
黑明燕,刘笑蓉,罗娅丽
(中南大学湘雅三医院儿科,长沙410013)
[摘要]目的:观察丹参酮ⅡA( Tanlla)对新生大鼠缺氧缺血性脑损伤(HIBD)脑皮质神经细胞
S897位点磷酸化的NMDA受体-1亚基( phospho-NR1S897)表达以及细胞内游离钙离子浓度([Ca2t]
的影响,探讨 TTANIIA对HIBD的保护作用机制。方法:7日龄新生SD大鼠240只随机分为正常对照组、
HIBD组及 HIBD+ Tanlla组,按Rice- Vannucci方法制作HIBD动物模型,时间点诜择HDB后3,6,12和
24h(各组各时间点n=10)。 TANIIA按1g/g每12h腹腔注射1次。应用Fura-2AM标记和日立F
4500型荧光担描仪测定[Ca2];。应用免疫荧光组化染色方法测定 phospho-NRlS897的表达。结果
(1)与正常对照组比较,HIBD组损伤侧脑细胞[Ca2+],绝对值和损伤侧与对侧的[Ca2】,比值明显升
高,各时间点的差异均具有统计学意义(P<0.05);与IIBD组比較,HIBD+ TANIA组各时间点的
[Ca2],升高的程度均有所减轻,其中24h损伤例与对側的[Ca2,比值較HBD组降低24.9%,差异
具有统计学意义(P<0.05)。(2)正常对照组大脑皮质有大量均匀分布的 phospho-NR1S897染色阳性
细胞。与正常对照组比较,HBD组各时间点损伤侧脑皮质区 phospho-NRIS897旧性表达细胞数和録色
荧光强度均明显降低,差异均具有统计学意义(P<0.05)。与HIBD组比较,HIBD+ Tanlla组各时间点、
的损伤例脑皮质区 phospho-NR1S897阳性表达细胞数和绿色荧光强度均升高,其中HIBD后3h和24h
的差异具有统计学意义(P<0.05)。结论: ITANIIA可以减轻HBD导致的脑皮质区 phospho-NR1S897表
达减弱的程度,并降低HIBD导致的[Cea2']升高的程度,提示 Tanlla对脑细胞的保护作用可能是通过
影响NMDA受体的表达,减少细胞内游离钙聚集而达到的
关键词]丹参酮IA;缺氧缺血;脑损伤; phospho-NR1S897;大鼠
D)OL:10.3969/j.issa.1672-7347.2010.09.007
Effect of Tanshinone IA on phosphorylated NMDA receptor
expression and intracellular free calcium concentration
in neonatal SD rats with hypoxic ischemic brain damage
HEI Mingyan, Llu Furong, LUO Yali
Department of Pediatrics, Third Xiangya Hopital, Central South University, Changsha 410013, China
Abstract: Objective To determine the effect of Tanshinone IIA(Tanl A) on the phosphory
lated NMDA receptor 1 at Serine 897 site (phospho-nr1 $897 and intracellular free calcium con-
收稿日期( Date of reception)2009-12-24
作者简介( Biography)黑明燕,博士,副主任医师,主要从事新生儿疾病研究。
通信作者(Correspondingauthor)黑明燕,E-mail:hemming_yane@yahoo.com.cn
基金项目( Foundation item)湖南省科技厅科研课题(2008FJ3157)。 This work was suported by the Department of Science and Technol-
ogy of Hunan Province, P. R. China(2008P 3157)
外参酮IL对新生大鼠缺氧缺血性脑损伤磷酸化NMDA受体1表达及细胞内游离钙浓度的影响黑明燕,等
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centration([Ca""] ) in neonatal SD rats with hypoxic ischemic brain damage(HIBD), and to e
plore the neuroprotective mechanism of TANJIA in HIBD. Methods Neonatal SD rats were randomly
divided into a normal control, and an HIBD and Tanlla HIBD group. Rice-vannucci method was
used for HIRD animal model. Time points were: 3, 6, 12, and 24 h after HIB(n=10 in eac
group at each time point ) Tanil A was intraperitoneally given at 1 ug/g every 12 h. Fura-2 AM was
used to mark the fluorescent calcium probe and Ca"'], was measured by a Hitachi F-4500 Fluores-
cence Spectrophometer. Fluorescent immunhisotichemical study was used for the expression of phos
PHO-NRI $897. Results (1 Compared with the normal control group, both the[ Ca"J, absolute
number and ipsi-/contra-lateral ratio were increased at cach time point with statistical significance(P
<0.05). Compared with the HIBD group, the Ca"1, in the HIBD +Tanlla group was de
creased at each time point. At 24 h after H]BD, the ipsi-/contra-lateral ratio of HIBD+Tanlia
group was 24. 9% less than that of FIBD group with statistical significance(P<0.05).(2)I
the normal control group, abundant phospho-nr 1 $897 positive cells were nicely distributed in the
cortex. Compared with the normal control group, at each time point, both the absolute numb