基因重组长效人促肾上腺皮质激素药理学活性

Pharmacological effects of gene recombinant longlasting human adrenocorticotropic hormone

目的为了提高促肾上腺皮质激素(adrenocorticotropic hormone,ACTH)的稳定性,针对ACTH中多种血液蛋白酶的作用位点,制备特定氨基酸位点突变(E5D、F7L、G10A及Y2S)基因重组长效人ACTH,分析比较哪一种点突变基因重组ACTH保留了天然野生型的生理活性。方法采用ELISA和q PCR等方法比较了4种突变型ATCH与野生型ACTH对体外培养的小鼠肾上腺皮质Y1细胞分泌糖皮质激素(glucocorticoids,GC)水平的影响,以及它们对ACTH合成的主要限速酶基因(STAR、P450-scc、3β-HSD)以及对STAR和P450-scc的合成发挥关键调节作用的类固醇合成因子1(SF-1)的转录水平的影响。结果 E5D及Y2S突变型ACTH对Y1细胞分泌GC以及对SF-1、STAR、P450-scc、3β-HSD等基因表达的影响与野生型ACTH比较,没有显著性差异。结论一级结构修饰的E5D和Y2S突变型ACTH的生理活性与野生型ACTH相当。

Objective To increase the stability of adrenocorticotropic hormone（ ACTH）,we prepared E5 D,F7L,G10 A and Y2 S mutant gene-recombinant long-lasting ACTH,based on analysis of consensus sequences of substrates of the proteases in the blood. The aim of the study is to compare which of the mutant ACTH candidates has pharmacological effects equivalent to that of wild type ACTH. Methods Using ELISA and q PCR method,we compared the effects of the mutant ACTHs and wild type ACTH on secretion of glucocorticoids（ GC） and transcription levels of the genes encoding enzymes critical for GC synthesis including STAR,P450-scc,3β-HSD,and steroidogenic factor 1（ SF-1） which plays a critical role in regulating the expression STAR and P450-scc. Results There was no significant difference between wild type ACTH and E5D-and Y2S-mutant ACTH in their effects on secretion of GC and transcription of the above genes. Conclusions The pharmacological effects of E5D-and Y2S-mutant ACTH is equivalent to those of wild type ACTH.