摘要
目的 研究吗啡对C6胶质瘤细胞嘌呤核苷酸合成代谢与分解代谢的影响。方法 吗啡 (10 μg/ml培养基 )作用于C6胶质瘤细胞 6h、12h、2 4h、4 8h、72h ;纳络酮 (1μmol/L)作用于C6胶质瘤细胞 1h后 ,加吗啡 (10 μg/ml)作用 2 4h。提取细胞总RNA ,采用反转录 聚合酶链反应 (RT PCR)方法检测次黄嘌呤 鸟嘌呤磷酸核糖转移酶 (HGPRT)及腺苷激酶 (AK)的基因转录产物 ;采用反转录 聚合酶链反应及Southern(RT PCR Southern)杂交方法检测黄嘌呤脱氧酶 (XD) /黄嘌呤氧化酶 (XO)基因转录产物。结果 C6胶质瘤细胞暴露于吗啡 6h ,12h ,2 4h ,4 8h ,HGPRT基因表达均明显降低 ;而C6胶质瘤细胞AK基因表达在暴露于吗啡 2 4h明显降低 ;HGPRT与AK基因表达又分别于吗啡作用胶质瘤细胞 72h和 4 8h回升 ;纳络酮不能阻断吗啡对HGPRT与AK基因表达降低的作用。吗啡作用于C6胶质瘤细胞与相应时段对照组相比 ,XD/XO基因转录产物均明显增多 ;纳络酮能够阻断吗啡对XD/XO基因表达增强的作用。结论 吗啡通过其他非 μ受体途径介导对C6胶质瘤细胞嘌呤核苷酸补救合成关键酶HGPRT与AK基因表达降低的作用 ;吗啡通过 μ受体途径介导对C6胶质瘤细胞嘌呤核苷酸分解代谢关键酶XD/XO基因表达增强的作用。
Objective To investigate the effect of morphine on catabolism and anabolism of purine nucleotide in c6 glioma cells. Methods C6 glioma cells were cultured and divided into 3 groups: 1) morphine group: morphine (10 μg/ml culture) was added for 6 h, 12 h, 24 h, 48 h, and 72 h; 2) morphine + naloxone group: naloxone (1 μmol/L) was added for 1 hour and then morphine (10 μg/ml) was added for 24 hours; and 3) control group: normal saline was used for 6, 12, 24, 48, and 72 hours. The C6 glioma cells were centrifuged. RT PCR was used to examine the gene transcripts of key enzymes of purine salvage way, hypoxanthine guanine phosphoribosyl transferase (HGPRT) and adenylate kinase (AK). RT PCR Southern blotting was used to examine the gene transcripts of key enzymes of purine salvage way, xanthine dehydrogenase (XD)/xanthine oxidase (XO) mRNA. Results Compared with that in the control group, the transcript of AK mRNA was significantly lower in the C6 cells treated with morphine for 24 hours, and began to re increase 48 hours after morphine treatment. The transcript of AK mRNA remained at a low level after treatment of naloxone for 1 hour and treatment of morphine for 24 hours. The levels of transcript of HGPRT mRNA were lower in the morphine group than in the control group at all time points after treatment. However, the level of transcript of HGPRT mRNA 72 hours after treatment was higher in the morphine group than in the control group. The level of transcript of HGPRT mRNA 24 hours after exposure to morphine in the naloxon2 + morphine group was still lower than in the control group. The levels of transcripts of XD/XO mRNA were significantly higher after exposure to morphine in comparison with those in the control group at all time points after treatment. However, the levels of XD/XO mRNA 24 hours after exposure to morphine in the naloxone + morphine group recovered to the normal level. Conclusion The downregulation effect of morphine on the gene expression of AK and HGPRT may not be mediated by μ receptor.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2003年第1期46-50,共5页
National Medical Journal of China
