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目的研究神经肽内皮素(ET)和心钠素(ANP)在细胞培养及体内实验中对促甲状腺激素释放激素受体(TRHR)基因表达的影响。方法Northern印迹和S1核酸酶保护实验检测TRHRmRNA量,新生肽链试验测定TRHR基因的转录。结果ET32h即增加胶质细胞TRHRmRNA量,6~8h达高峰,增加3倍以上(3.0±0.52);相反,ANP2h即降低TRHRmRNA,6~8h达最低值,并逆转60~80%因血浆所增高的TRHRmRNA。ET3在6h时增加TRHR基因的转录达2倍(nuclearrunon试验),而ANP对TRHR基因的转录无明显变化。体内实验结果显示ET3和ANP对TRHRmRNA影响与体外实验相似。结论ET3可能通过调节TRHR基因的表达而影响TRHRmRNA,而ANP则可能影响TRHRmRNA的降解而降低mRNA的水平。
Objective To investigate the effect of neuropeptide endothelin (ET) and atrial natriuretic peptide (ANP) on the gene expression of thyrotropin releasing hormone receptor (TRHR) in cell culture and in vivo experiments. Methods The amount of TRHR mRNA was detected by Northern blotting and S1 nuclease protection assay, and the TRHR gene transcription by nascent peptide chain assay. Results The amount of TRHR mRNA in glial cells was increased at 32 h after ET (P <0.05), reaching a peak at 6-8 h (3.0 ± 0.52). On the contrary, TRP mRNA decreased at ANP2 h and reached the lowest at 6-8 h, 80% increase in plasma TRHR mRNA. ET 3 increased TRHR gene transcription up to 2 times (nuclearrunon test) at 6h, while ANP had no obvious change on TRHR gene transcription. In vivo experimental results show that ET 3 and ANP on TRHRmRNA and in vitro experiments similar. Conclusion ET3 may affect TRHR mRNA by regulating the expression of TRHR gene, while ANP may affect TRHR mRNA degradation and decrease mRNA level.