复方党参对高原低氧大鼠脑细胞凋亡相关基因表达谱影响的分析  

Expression analysis of Compound Codonopsis pilosula Nanufeldt on the hypoxic cerebral apoptosis and its mechanism

作　　者：刘刊[1] 聂鸿靖[1] 马子敏[2] 张东祥[1] 张波[1] 范明[2] 尹昭云[1] 谢印芝[1] 

机构地区：[1]军事医学科学院卫生学环境医学研究所,天津300050 [2]军事医学科学院基础医学研究所,北京100850

出　　处：《中国现代应用药学》2004年第5期345-349,共5页Chinese Journal of Modern Applied Pharmacy

基　　金：国家自然科学基金资助项目 ( 3 99790 90 8)

摘　　要：目的　研究复方党参对模拟高原低氧条件下大鼠脑神经细胞凋亡基因表达谱的作用 ,从中药与生物体基因网络相互作用的角度 ,探讨复方党参的作用机制。方法　将Wistar大鼠置于模拟海拔 70 0 0m的低压氧舱建立高原低氧大鼠脑细胞凋亡模型 ,于上舱前 7d开始以复方党参液灌胃给药。提取空白对照组、低氧 8d对照组、常氧复方党参组及低氧 8d复方党参组大鼠脑组织总RNA ,经逆转录合成cDNA探针 ,用 [α 3 2 P]标记探针 ,各自与AtlascDNAArray膜杂交 ,经严格的冲洗后用磷屏检cDNA表达水平。获得的基因表达谱数据集采用自组图 (SOMs)法分析。结果　结果表明在模拟高原低氧条件下 ,导致神经细胞凋亡的基因 (p5 3、bax、bad、bok、Hsp6 0 )聚为一类 (即表达行为相似 )。而具有抗凋亡功能的基因 (bcl 2、bcl xl、Hsp70、Hsp2 7、Hsp90、Hsc70、Rad)聚为一类。 结论　低氧可以启动细胞凋亡的线粒体途径 ,导致细胞死亡 ;而复方党参以凋亡相关基因网络中的多种成分为靶点 。OBJECTIVE The purpose of this study was to investigate the effect of Compound Codonopsis pilosula Nanufeldt(CCPN)on the hypoxic neuronal apoptosis and elucidate its mechanisms using cDNA microarray techniques. METHOD Wistar rats (body weight 180～220g) were randomly divided into normoxia and hypoxia for 8 days group, each group was subdivided into CCPN group and control group. CCPN was given to the rats by gastric incubation for 7 days before putting into the hypobaric chamber at a simulated altitude of 7000m for 5 hours each day. Then the rats were decapitated. Total RNA of different groups was isolated, then probe mixtures were synthesized by reverse-transcribing each RNA population using the cDNA synthesis (CDS) primer mix and dATP. Each radioactively labeled probe mix was then hybridized to separate Atlas Arrays. After a high stringency wash, the hybridization pattern was analyzed by phosphorimaging. The expression patterns of genes were interpreted by self-organizing maps (SOMs). RESULTS We found that proapoptosis genes such as p53, bax, bad, bok and Hsp60 were organized into the same cluster, the expression levels of these genes elevated under the circumstance of hypoxia, but downregulated in the CCPN group, while antiapoptosis genes bcl-2, bcl-xl and genes encoded Hsp70, Hsp90, Hsp27, Hsc70, Ras associated protein were grouped together and upregulated by this medicine. These results indicated that CCPN can reduce the ratio of expression between bax/bcl-2 or bax/bcl-xl to prevent apoptosis. Ras-MAP signaling pathway may also involve in this process.CONCLUSION Hypoxia could trigger the internal (mitochondrial) pathway of apoptosis though upregulating proapoptotic proteins as P53, Bax, Bad, Hsp60 and resulted in apoptosis. CCPN possesses an antiapoptosis function. There are at least two mechanisms of its action. The first mechanism involves regulating the expression level of Bcl-2, Bax, Hsp70 and increasing the ratio between Bcl-2/Bax, which determines apoptosis or survive; while the second one is CCPN u

关 键 词：复方党参 高原低氧 神经细胞 凋亡 微阵列 

分 类 号：B594.3[哲学宗教—外国哲学] R285[医药卫生—中药学]