吗啡对PC12细胞嘌呤核苷酸代谢相关酶基因表达的影响

doi:国家博士后基金资助课题(200435566)；吉林

吗啡对PC12细胞嘌呤核苷酸代谢相关酶基因表达的影响

袁海英¹，刘英鹰² ，刘剑凯^1＊，洪敏¹

1. 吉林大学基础医学院生物化学与分子生物学教研室，吉林长春 130021；2. 吉林市第二中心医院中医科，吉林吉林132001

收稿日期:2005-09-02 修回日期:1900-01-01 出版日期:2006-11-28 发布日期:2006-11-28
通讯作者: 洪敏

Effect of morphine on gene expression of enzymes related to purine nucleotides metabolism in PC12 cells

YUAN Hai-yin¹，LIU Ying-ying²，LIU Jian-kai^1＊，HONG Min¹ 

1. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China; 2.Department of Traditional Chinese Medicine, Second Central Hospital of Jilin City, Jilin 132001, China

Received:2005-09-02 Revised:1900-01-01 Online:2006-11-28 Published:2006-11-28
Contact: LIU Jian-kai

摘要/Abstract

摘要： 目的：研究吗啡对神经细胞模型PC12嘌呤核苷酸补救合成酶与分解代谢关键酶基因表达的影响。方法：采用逆转录聚合酶链反应（RT-PCR）方法检测暴露于吗啡不同时间的PC12细胞次黄嘌呤-鸟嘌呤磷酸核糖转移酶（HGPRT）、腺苷激酶（AK）和腺苷脱氨酶（AD A）mRNA的表达水平。结果：与相应时段对照组相比，PC12细胞暴露于吗啡12及24 h时，HGPRT mRNA水平均明显增高(P<0.05)，作用48 h时HGPRT mRNA水平显著降低(P<0.05)，72 h后HGPRT mRNA水平再次升高(P<0.05)；吗啡处理的PC12细胞于12和24 h时，AK mRNA水平均明显降低(P<0.05)，作用48 h时AK mRNA水平升高(P<0.05)，72 h后AK mRNA水平恢复正常；ADA mRNA水平均表现为轻度降低，12 h差异有显著性(P<0.05)。结论：吗啡影响神经细胞嘌呤核苷酸代谢，使细胞内腺苷酸及腺苷水平增高，这可能是吗啡依赖和耐受形成的机理之一。

关键词: PC12细胞, 次黄嘌呤磷酸核糖转移酶, 腺苷激酶, 腺苷脱氨酶

Abstract: Objective To study the effect of morphine on gene expression of enzymes involved in salvage and catabolism pathways of purine nucleotides metabolism in nerve cells. Methods PC12 cells were cultivated and divided into morphine treated groups which were treated with morphine (10 mg•L^-1 culture) for 12, 24, 48,72 h, and control groups which were treated with normal saline for 12, 24, 48 and 72 h. Total RNA of PC12 cells was isolated. HGPRT, AK, ADK mRNA levels were determined by using the reverse transcription polymerase chain reaction (RT-PCR);the β-actin mRNA expression was used as an internal control. Results As compared with corresponding control groups, HGPRT mRNA levels in PC12 cells were increased significantly after 12 and 24 h treatment with morphine(P<0.05), became lower after 48 h and became higher again after 72 h (P<0.05). AK mRNA levels in PC12 cells were decreased significantly after 12 and 24 h treatment with morphine(p<0.05), become higher after 48 h (P<0.05), and were normal after 72 h. ADA mRNA levels were decreased in morphine treated PC12 cells in all the given periods with significant difference in 12 h (P<0.05). Conclusion Morphine may affect purine nucleotide metabolism of nerve cells, increase the adenylate and adenosine levels, which may be one of the mechanism of morphine dependence and tolerance.

Key words: PC12 cells, hypoxanthine phosphoribosyltransferase, adenosine kinase, adenosin deaminase

中图分类号:

R786

袁海英，刘英鹰，刘剑凯，洪敏. 吗啡对PC12细胞嘌呤核苷酸代谢相关酶基因表达的影响[J]. J4, 2006, 32(6): 1037-1040.

YUAN Hai-yin，LIU Ying-ying，LIU Jian-kai，HONG Min . Effect of morphine on gene expression of enzymes related to purine nucleotides metabolism in PC12 cells[J]. J4, 2006, 32(6): 1037-1040.

[1]	徐姗姗, 朴美花, 王艳姝, 刘楠, 裴爱月, 冯春生. 异氟醚对Aβ25-35诱导大鼠PC12细胞氧化应激损伤的影响及海藻糖的保护作用[J]. 吉林大学学报(医学版), 2015, 41(02): 207-212.
[2]	邹泳国，周春燕，朴美花，刘楠，岳云，冯春生. 异氟醚对β淀粉样蛋白诱导的大鼠PC12细胞凋亡和内质网应激的影响及其机制[J]. 吉林大学学报(医学版), 2014, 40(02): 224-228.
[3]	王伟，朴美花，王艳姝，刘楠，岳云，冯春生. 异氟醚对Aβ25-35诱导大鼠PC12细胞自噬和凋亡的影响[J]. 吉林大学学报(医学版), 2014, 40(01): 49-54.
[4]	姜恩平, 李红, 杨世杰. 蒺藜皂苷对氧化应激PC12细胞内NF-κB水平的影响及其机制[J]. J4, 2010, 36(6): 1035-1038.
[5]	崔佳乐, 何海涛, 洪新雨, 邓志会, 赵丽艳, 洪敏. 嘌呤核苷酸对海洛因依赖及戒断大鼠睾丸和附睾组织中腺苷脱氨酶活性及基因表达的影响[J]. J4, 2010, 36(2): 291-294.
[6]	李昆, 李鸿梅, 何海涛, 刘剑凯, 洪敏. 嘌呤核苷酸补偿对海洛因依赖大鼠嘌呤核苷酸分解代谢的影响[J]. J4, 2009, 35(5): 837-840.
[7]	姜恩平, 吴金义, 陈建光. 北五味子总木脂素对H₂O₂诱导PC12细胞凋亡的影响[J]. J4, 2009, 35(3): 466-469.
[8]	姜恩平，李红，纪影实，杨世杰. 蒺藜总皂苷对H₂O₂诱导PC12细胞凋亡的保护作用[J]. J4, 2008, 34(2): 183-186.
[9]	王秀力，李忱，刘佳梅，陈东. 维甲酸对诱导PC12细胞向神经元样细胞分化的影响[J]. J4, 2008, 34(2): 187-190.
[10]	孙婷，何海涛，阚慕洁，张连芝，洪敏. 嘌呤核苷酸减弱吗啡抑制的PC12细胞嘌呤核苷酸合成代谢[J]. J4, 2007, 33(6): 971-973.
[11]	王秀力，陈东，刘佳梅. NGF诱导PC12细胞向神经元的分化[J]. J4, 2007, 33(5): 827-830.
[12]	张海娜，胡国华，陈秋惠，孙冬，刘彩霞，孙亚娟. 帕金森病细胞模型的建立及鱼藤酮对多巴胺能神经元的毒性作用[J]. J4, 2007, 33(5): 811-814.
[13]	韩威, 宫萍, 常明, 张瑜, 张颖, 王秋艳, 胡轶虹,胡林森. 鱼藤酮对PC12细胞毒性作用的机制[J]. J4, 2007, 33(1): 98-101.
[14]	张磊, 常明, 徐卉, 侯澍, 李红杰, 朴燕洁, 胡林森. PC12细胞蛋白质组学研究技术及二维电泳图谱的建立[J]. J4, 2006, 32(6): 1112-1115.
[15]	房绍宽，吴江，周春奎. 新型抗精神失常药万拉法辛和奥氮平对无血清条件下PC12细胞的保护作用[J]. J4, 2006, 32(3): 480-482.