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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 study the effects of morphine on the metabolism and catabolism of purine nucleotides in C6 glioma cells. METHODS: C6 glioma cells were treated with morphine (10μg / ml) for 6h, 12h, 24h, 48h, 72h respectively. After 1 mol / L naloxone treatment for 1h, 10 μg / ml) for 24 h. The total cellular RNA was extracted and the transcripts of hypoglyceral guanine phosphoribosyltransferase (HGPRT) and adenosine kinase (AK) were detected by reverse transcriptase-polymerase chain reaction (RT PCR) (XD) / xanthine oxidase (XO) gene transcripts were detected by Southern blot hybridization and Southern blotting. Results The expression of HGPRT gene was significantly decreased in C6 glioma cells exposed to morphine for 6h, 12h, 24h, 48h, while the expression of AK gene in C6 glioma cells was significantly decreased 24 h after exposure to morphine. The expressions of HGPRT and AK gene Respectively, in morphine-induced glioma cells 72h and 48h recovery; naloxone can not block morphine on HGPRT and AK gene expression decreased role. Compared with the control group, the transcription of XD / XO gene was significantly increased in morphine treated C6 glioma cells. Naloxone blocked the enhancement of XD / XO gene expression by morphine. Conclusion Morphine mediates the decrease of HGPRT and AK gene expression in C6 glioma cells through other non-mu receptor pathway. Morphine mediates the decrease of purine nucleus in C6 glioma cells through μ receptor pathway Enhancement of XD / XO gene expression, a key enzyme in catabolism.