垂体GH腺瘤中GHS-R启动子的功能研究
|
摘要
第一部分GHS-R启动子区缺失突变体的克隆及鉴定
目的构建一系列GHS-R基因5’侧翼启动子区不同长度缺失突变体的萤火虫荧光素酶报告基因增强子载体。
方法以原代培养的人GH腺瘤细胞基因组DNA为模版,PCR扩增GHS-R基因5’侧翼启动子区不同长度的片段,再定向克隆到pGL3-Enhancer载体中。结果通过酶切鉴定和基因测序,证明成功地构建了pGL3-Enhancer A~F克隆重组体。
结论为体外研究GHS-R基因的转录调节提供了新的手段,为下一步在垂体GH腺瘤中GHS-R基因启动子的干预研究奠定了基础。
第二部分GHS-R启动子区初步功能分析的实验研究
目的利用构建好的含有人GHS-R启动子区不同长度的缺失突变体的重组荧光素酶报道载体,对GHS-R启动子区的结构和功能进行初步的分析。
方法①将含有人GHS-R最短启动子区片段的pGL3-Enhancer-A和最长启动子区片段的pGL3-Enhancer-F质粒DNA分别转染293细胞、95D细胞和GH3细胞。②用双荧光素酶报道系统分别检测启动子活性,判断GHS-R启动子的细胞特异性。③将含有不同长度的人GHS-R启动子区缺失突变体的荧光素酶报道载体质粒转染GH3细胞。④用双荧光素酶报道系统检测各个重组子的启动子活性,筛选GHS-R启动子区重要的转录调控区域。
结果①转染pGL3-Enhancer-A质粒DNA后,各组细胞间启动子相对活性比较,统计学上无显著性差异(p>0.05);转染pGL3-Enhancer-F质粒DNA后,GH3细胞组启动子活性较293细胞组和95D细胞组有明显提高(p<0.01)。②含有不同长度的人GHS-R启动子区缺失突变体的荧光素酶报道载体质粒转染GH3细胞后启动子活性分析结果为:pGL3-Enhancer-B相对活性(8.4±0.86)与pGL3-Enhancer-A相对活性(2.3±0.21)相比,有显著升高,两组比较p<0.01;pGL3-Enhancer-C相对活性(5.2±0.30)与pGL3-Enhancer-B相对活性(8.4±0.86)相比,有明显下降(p<0.05);pGL3-Enhancer-D(10.6±0.54)相对活性与pGL3-Enhancer-C(5.2±0.30)相对活性相比,有明显升高,两组比较p<0.05;pGL3-Enhancer-E(14.1±0.74)相对活性与pGL3-Enhancer-D相对活性(10.6±0.54)相比,有明显升高,两组比较p<0.05;pGL3-Enhancer-F相对活性(12.9±0.23)与pGL3-Enhancer-E(14.1±0.74)相比,有所降低,但该变化无统计学意义(p>0.05)。
结论人GHS-R启动子(-1008~+10)能在GH3细胞中特异性表达活性,而在293细胞和95D细胞中无明显作用;人GHS-R启动子区-168~+10区域之间无作用明显的启动子调控元件;在人GHS-R启动子上游-254~-168之间,-625~-355之间,-910~-625之间存在正性调控区域;而在-355~-254之间存在负性调控区域。
第三部分GHS-R启动子区激素相关顺式作用元件的实验研究
目的研究GHS-R启动子的激素调控,明确GHS-R启动子区激素相关顺式作用元件的基因座位。
方法①用不同试剂干预有启动子活性pGL3-Enhancer-F质粒DNA瞬时转染GH3细胞。②双荧光素酶报告基因系统检测启动子活性,判定有效的干预试剂。③在有或无氢化可的松、T3和雌二醇干预下,pGL3-Enhancer-A~F载体及空载体转染GH3细胞。④用双荧光素酶报道系统检测各个重组子的启动子活性,判定启动子区激素相关顺式作用元件的特异性序列。
结果①氢化可的松作用组,干预前后转pGL3-Enhancer-F载体较转pGL3-Enhancer空载体相对荧光素酶活性比值明显下降; T3作用组和雌二醇作用组,干预前后转pGL3-Enhancer-F载体较转pGL3-Enhancer空载体相对荧光素酶活性比值明显上升;余试剂作用组干预前后相对荧光素酶活性比值在统计学上无明显差异。②在有无氢化可的松作用时,pGL3-Enhancer-C组启动子相对活性比值(0.7±0.10)与pGL3-Enhancer-B组启动子相对活性比值(1.2±0.17)相比,有显著下降;在有无T3作用时,pGL3-Enhancer-B组启动子相对活性比值(1.8±0.22)与pGL3-Enhancer-A组启动子相对活性比值(1.2±0.16)相比,有明显地提高;在有无雌二醇作用时,pGL3-Enhancer-B组启动子相对活性比值(2.0±0.17)与pGL3-Enhancer-A组启动子相对活性比值(1.2±0.11)相比,有明显地提高。
结论在人GHS-R启动子区(-355~-254)存在氢化可的松相关的顺式作用元件,且能被氢化可的松负性调控;在GHS-R启动子区(-254~-168)存在T3和雌二醇相关的顺式作用元件,且分别能被T3和雌二醇所正性调控。
PartⅠConstructing and Identifying the GHS-R Promoter Deletion Mutants
Objective: To construct a series of firefly luciferase report gene enhancer vectors for 5’flanking promoter region of GHS-R gene.
Methods: A series of DNA fragments of 5’flanking promoter region of GHS-R gene were amplified from the genomic DNA of primary cultured human pituitary somatotrophinomas cells in PCR. The PCR products were then cloned into the pGL3-Enhancer vector.
Results: Restriction enzymes digestion and nucleotide sequencing confirmed that the recombinant plasmids pGL3-Enhancer A~F had been constructed.
Conclusion: A new methods was provided to study the transcription regulation of GHS-R gene in vitro. A foundation for further study of GHS-R gene promoter in human pituitary somatotrophinomas was established.
PartⅡPrimary Function Analysis of GHS-R Promoter
Objective: To analyze the structure and function of GHS-R promoter region primarily, using the constructed recombinant reporter gene promoter-pGl3-enhancer vectors containing different length of GHS-R promoter deletion mutants.
Methods:①The recombinant plasmid pGL3-Enhancer-A containg the shortest length of human GHS-R promoter deletion mutants and pGL3-Enhancer-F containg the longest length of human GHS-R promoter deletion mutants were transfected into 293 cell, 95D cell and GH3 cell with the liposomes.②The promoter activities of different recombinant plasmids were detected by the Dual Luciferase Reporter Assay System, so the cell specificity of GHS-R promoter was identified.③The recombinant plasmids containg different length of human GHS-R promoter deletion mutants were transfected into GH3 cell.④The promoter activities of different recombinant plasmids were detected by the Dual Luciferase Reporter Assay System, then the important transcriptional regulation region of GHS-R promoter were identified.
Results:①After transfected pGL3-Enhancer-A plasmid, we found that there was not significant difference between the promoter activities of various cell in statistics(p>0.05). After transfected pGL3-Enhancer-F plasmid, we found that the promoter activity of GH3 cell increased markedly compared with 293 cell and 95D cell.②According to the Dual Luciferase Reporter Assay System, we found that the promoter activities increased markedly between pGL3-Enhancer-B(8.4±0.86)and pGL3-Enhancer-A(2.3±0.21),p<0.01; pGL3-Enhancer-D(10.6±0.54) and pGL3-Enhancer-C(5.2±0.30), p<0.05; pGL3-Enhancer-E(14.1±0.74)and pGL3-Enhancer-D(10.6±0.54), p<0.05, and also observed that the promoter activities declined significantly between pGL3-Enhancer-C(5.2±0.30)and pGL3-Enhancer-B(8.4±0.86), p<0.05, the promoter activities declined between pGL3-Enhancer-F(12.9±0.23)and pGL3-Enhancer-E(14.1±0.74), but there was not significant difference, p>0.05.
Conclusion: Human GHS-R promoter(-1008~+10)has specific expression activity in GH3 cell; There were not regulation elements between -168~+10 in the upstream of the GHS-R promoter; there was a positive transcriptional regulation region from -254 to -168, -625 to -355, -910 to -625 respectively, and a negative transcriptional regulation region between -355 to -254 in the upstream of the human GHS-R promoter.
PartⅢExperimental study of hormone related cis-element in the GHS-R promoter region
Objective: To investigated the hormone regulation of GHS-R promoter, and identify the specific sequences of hormone related cis-element in the GHS-R promoter region.
Methods:①Using various agents interfered the process that pGL3-Enhancer-F plasmid was transiently transfected GH3 cells.②We analyzed the effects of various agents by detecting the activity of promoter.③Plasmids containing progressively decreasing amount of GHS-R promoter region upstream of the luciferase gene were transiently transfected into GH3 cells, with or without hormonal treatment.④The promoter activities of different recombinant plasmids were detected by the Dual Luciferase Reporter Assay System, so the specific sequences of the hormone related cis-elements in the GHS-R promoter region were identified.
Results:①Treatment with hydrocortisone significantly inhibited the promoter activities of pGL3-Enhancer-F; treatment with T3 orβ-estradiol significantly enhanced the pGL3-Enhancer-F; and treatment with forskolin, somatostatin, TPA and IGF-1 did not significantly influence activity of the GHS-R promoter region analyzed.②With or without hydrocortisone , we observed that the ratio of relative activity declined significantly between pGL3-Enhancer-C(0.7±0.10)and pGL3-Enhancer-B(1.2±0.17); with or without T3, we found that ratio of relative activity increased markedly between pGL3-Enhancer-B(1.8±0.22)and pGL3-Enhancer-A(1.2±0.16); with or withoutβ-estradiol, we observed that ratio of relative activity increased markedly between pGL3-Enhancer-B(2.0±0.17)and pGL3-Enhancer-A(1.2±0.11).
Conclusion: A negative glucocorticoid-responsive element may be located in the GHS-R promoter region between -355 and -254; Positive thyroid and estrogen responsive elements may be located in the GHS-R promoter region between -254 and -168.
目的构建一系列GHS-R基因5’侧翼启动子区不同长度缺失突变体的萤火虫荧光素酶报告基因增强子载体。
方法以原代培养的人GH腺瘤细胞基因组DNA为模版,PCR扩增GHS-R基因5’侧翼启动子区不同长度的片段,再定向克隆到pGL3-Enhancer载体中。结果通过酶切鉴定和基因测序,证明成功地构建了pGL3-Enhancer A~F克隆重组体。
结论为体外研究GHS-R基因的转录调节提供了新的手段,为下一步在垂体GH腺瘤中GHS-R基因启动子的干预研究奠定了基础。
第二部分GHS-R启动子区初步功能分析的实验研究
目的利用构建好的含有人GHS-R启动子区不同长度的缺失突变体的重组荧光素酶报道载体,对GHS-R启动子区的结构和功能进行初步的分析。
方法①将含有人GHS-R最短启动子区片段的pGL3-Enhancer-A和最长启动子区片段的pGL3-Enhancer-F质粒DNA分别转染293细胞、95D细胞和GH3细胞。②用双荧光素酶报道系统分别检测启动子活性,判断GHS-R启动子的细胞特异性。③将含有不同长度的人GHS-R启动子区缺失突变体的荧光素酶报道载体质粒转染GH3细胞。④用双荧光素酶报道系统检测各个重组子的启动子活性,筛选GHS-R启动子区重要的转录调控区域。
结果①转染pGL3-Enhancer-A质粒DNA后,各组细胞间启动子相对活性比较,统计学上无显著性差异(p>0.05);转染pGL3-Enhancer-F质粒DNA后,GH3细胞组启动子活性较293细胞组和95D细胞组有明显提高(p<0.01)。②含有不同长度的人GHS-R启动子区缺失突变体的荧光素酶报道载体质粒转染GH3细胞后启动子活性分析结果为:pGL3-Enhancer-B相对活性(8.4±0.86)与pGL3-Enhancer-A相对活性(2.3±0.21)相比,有显著升高,两组比较p<0.01;pGL3-Enhancer-C相对活性(5.2±0.30)与pGL3-Enhancer-B相对活性(8.4±0.86)相比,有明显下降(p<0.05);pGL3-Enhancer-D(10.6±0.54)相对活性与pGL3-Enhancer-C(5.2±0.30)相对活性相比,有明显升高,两组比较p<0.05;pGL3-Enhancer-E(14.1±0.74)相对活性与pGL3-Enhancer-D相对活性(10.6±0.54)相比,有明显升高,两组比较p<0.05;pGL3-Enhancer-F相对活性(12.9±0.23)与pGL3-Enhancer-E(14.1±0.74)相比,有所降低,但该变化无统计学意义(p>0.05)。
结论人GHS-R启动子(-1008~+10)能在GH3细胞中特异性表达活性,而在293细胞和95D细胞中无明显作用;人GHS-R启动子区-168~+10区域之间无作用明显的启动子调控元件;在人GHS-R启动子上游-254~-168之间,-625~-355之间,-910~-625之间存在正性调控区域;而在-355~-254之间存在负性调控区域。
第三部分GHS-R启动子区激素相关顺式作用元件的实验研究
目的研究GHS-R启动子的激素调控,明确GHS-R启动子区激素相关顺式作用元件的基因座位。
方法①用不同试剂干预有启动子活性pGL3-Enhancer-F质粒DNA瞬时转染GH3细胞。②双荧光素酶报告基因系统检测启动子活性,判定有效的干预试剂。③在有或无氢化可的松、T3和雌二醇干预下,pGL3-Enhancer-A~F载体及空载体转染GH3细胞。④用双荧光素酶报道系统检测各个重组子的启动子活性,判定启动子区激素相关顺式作用元件的特异性序列。
结果①氢化可的松作用组,干预前后转pGL3-Enhancer-F载体较转pGL3-Enhancer空载体相对荧光素酶活性比值明显下降; T3作用组和雌二醇作用组,干预前后转pGL3-Enhancer-F载体较转pGL3-Enhancer空载体相对荧光素酶活性比值明显上升;余试剂作用组干预前后相对荧光素酶活性比值在统计学上无明显差异。②在有无氢化可的松作用时,pGL3-Enhancer-C组启动子相对活性比值(0.7±0.10)与pGL3-Enhancer-B组启动子相对活性比值(1.2±0.17)相比,有显著下降;在有无T3作用时,pGL3-Enhancer-B组启动子相对活性比值(1.8±0.22)与pGL3-Enhancer-A组启动子相对活性比值(1.2±0.16)相比,有明显地提高;在有无雌二醇作用时,pGL3-Enhancer-B组启动子相对活性比值(2.0±0.17)与pGL3-Enhancer-A组启动子相对活性比值(1.2±0.11)相比,有明显地提高。
结论在人GHS-R启动子区(-355~-254)存在氢化可的松相关的顺式作用元件,且能被氢化可的松负性调控;在GHS-R启动子区(-254~-168)存在T3和雌二醇相关的顺式作用元件,且分别能被T3和雌二醇所正性调控。
PartⅠConstructing and Identifying the GHS-R Promoter Deletion Mutants
Objective: To construct a series of firefly luciferase report gene enhancer vectors for 5’flanking promoter region of GHS-R gene.
Methods: A series of DNA fragments of 5’flanking promoter region of GHS-R gene were amplified from the genomic DNA of primary cultured human pituitary somatotrophinomas cells in PCR. The PCR products were then cloned into the pGL3-Enhancer vector.
Results: Restriction enzymes digestion and nucleotide sequencing confirmed that the recombinant plasmids pGL3-Enhancer A~F had been constructed.
Conclusion: A new methods was provided to study the transcription regulation of GHS-R gene in vitro. A foundation for further study of GHS-R gene promoter in human pituitary somatotrophinomas was established.
PartⅡPrimary Function Analysis of GHS-R Promoter
Objective: To analyze the structure and function of GHS-R promoter region primarily, using the constructed recombinant reporter gene promoter-pGl3-enhancer vectors containing different length of GHS-R promoter deletion mutants.
Methods:①The recombinant plasmid pGL3-Enhancer-A containg the shortest length of human GHS-R promoter deletion mutants and pGL3-Enhancer-F containg the longest length of human GHS-R promoter deletion mutants were transfected into 293 cell, 95D cell and GH3 cell with the liposomes.②The promoter activities of different recombinant plasmids were detected by the Dual Luciferase Reporter Assay System, so the cell specificity of GHS-R promoter was identified.③The recombinant plasmids containg different length of human GHS-R promoter deletion mutants were transfected into GH3 cell.④The promoter activities of different recombinant plasmids were detected by the Dual Luciferase Reporter Assay System, then the important transcriptional regulation region of GHS-R promoter were identified.
Results:①After transfected pGL3-Enhancer-A plasmid, we found that there was not significant difference between the promoter activities of various cell in statistics(p>0.05). After transfected pGL3-Enhancer-F plasmid, we found that the promoter activity of GH3 cell increased markedly compared with 293 cell and 95D cell.②According to the Dual Luciferase Reporter Assay System, we found that the promoter activities increased markedly between pGL3-Enhancer-B(8.4±0.86)and pGL3-Enhancer-A(2.3±0.21),p<0.01; pGL3-Enhancer-D(10.6±0.54) and pGL3-Enhancer-C(5.2±0.30), p<0.05; pGL3-Enhancer-E(14.1±0.74)and pGL3-Enhancer-D(10.6±0.54), p<0.05, and also observed that the promoter activities declined significantly between pGL3-Enhancer-C(5.2±0.30)and pGL3-Enhancer-B(8.4±0.86), p<0.05, the promoter activities declined between pGL3-Enhancer-F(12.9±0.23)and pGL3-Enhancer-E(14.1±0.74), but there was not significant difference, p>0.05.
Conclusion: Human GHS-R promoter(-1008~+10)has specific expression activity in GH3 cell; There were not regulation elements between -168~+10 in the upstream of the GHS-R promoter; there was a positive transcriptional regulation region from -254 to -168, -625 to -355, -910 to -625 respectively, and a negative transcriptional regulation region between -355 to -254 in the upstream of the human GHS-R promoter.
PartⅢExperimental study of hormone related cis-element in the GHS-R promoter region
Objective: To investigated the hormone regulation of GHS-R promoter, and identify the specific sequences of hormone related cis-element in the GHS-R promoter region.
Methods:①Using various agents interfered the process that pGL3-Enhancer-F plasmid was transiently transfected GH3 cells.②We analyzed the effects of various agents by detecting the activity of promoter.③Plasmids containing progressively decreasing amount of GHS-R promoter region upstream of the luciferase gene were transiently transfected into GH3 cells, with or without hormonal treatment.④The promoter activities of different recombinant plasmids were detected by the Dual Luciferase Reporter Assay System, so the specific sequences of the hormone related cis-elements in the GHS-R promoter region were identified.
Results:①Treatment with hydrocortisone significantly inhibited the promoter activities of pGL3-Enhancer-F; treatment with T3 orβ-estradiol significantly enhanced the pGL3-Enhancer-F; and treatment with forskolin, somatostatin, TPA and IGF-1 did not significantly influence activity of the GHS-R promoter region analyzed.②With or without hydrocortisone , we observed that the ratio of relative activity declined significantly between pGL3-Enhancer-C(0.7±0.10)and pGL3-Enhancer-B(1.2±0.17); with or without T3, we found that ratio of relative activity increased markedly between pGL3-Enhancer-B(1.8±0.22)and pGL3-Enhancer-A(1.2±0.16); with or withoutβ-estradiol, we observed that ratio of relative activity increased markedly between pGL3-Enhancer-B(2.0±0.17)and pGL3-Enhancer-A(1.2±0.11).
Conclusion: A negative glucocorticoid-responsive element may be located in the GHS-R promoter region between -355 and -254; Positive thyroid and estrogen responsive elements may be located in the GHS-R promoter region between -254 and -168.
引文
1 Asthagiri A, Lopes MB. Neuropathological considerations of pituitary adenomas. Front Horm Res, 2006, 34: 206-235.
2 Pong S, Chaung LP, Dean CD, et al. Identification of a new G-protein-link receptor for growth hormone secretagogues. Mol Endocrinol, 1996, 10: 57-61.
3 Howard AD, Feighner SD, Cully DF, et al. A receptor in pituitary and hypothalamus that functions in growth hormone release. Science, 1996, 273: 974-977.
4 Simith RG. Development of growth hormone secretagogues. Endocr Rev, 2005, 26 (3): 346-360.
5 Kamegai J, Tamura H, Shimizu T, et al. The role of pituitary ghrelin in growth hormone (GH) secretion: GH-releasinghormone-dependent regulation of pituitary ghrelin gene expression and peptide content. Endocrinology, 2004, 145(8): 3731-3738.
6 Volante M, Allia E, Gugliotta P, et al. Expression of ghrelin and of the GH secretagogue receptor by pancreatic islet cells and related endocrine tumors. J Clin Endocrinol Metab, 2002, 87: 1300-1308.
7 Penny L, Adrian C, Lisa K. The potential autocrine/paracrine roles of ghrelin and its receptor in hormone-dependent cancer. Cytokine & Growth Factor Reviews, 2003, 14: 113-122.
8 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
9 Yan M, Jones ME, Hernandez M, et al. Oestrogen replacement in vivo rescues the dysfunction of pituitary somatotropes in ovariectomised aromatase knockout mice. Neuroendocrinology, 2005, 81: 158-166.
10 Chen C. Growth hormone secretagogue actions on the pituitary gland: multiple receptors for multiple ligands? Clin Exp Pharmacol Physiol, 2000, 27: 323-329.
11 Katayama M, Nogami H, Nishiyama J, et al. Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland. Neuroendocrinology, 2000, 72: 333-340.
12 Kamegai J, Wakabayashi I, Miyamoto K, et al. Growth hormone-dependent regulation of pituitary GH secretagogue receptor (GHS-R) mRNA levels in the spontaneous dwarf Rat. Neuroendocrinology, 1998, 68: 312-318.
1 Bertherat J. Cloning of the growth hormone secretagogues receptor cDNA: new evidence for a third endocrine pathway controlling growth hormone release. Eur J Endocrinol, 1997, 136: 37-38.
2 Smith RG. Development of growth hormone secretagogues. Endocr Rev, 2005, 26: 346-360.
3 Skinner MM, Nass R, Lopes B, et al. Growth hormone secretagogue receptor expression in human pituitary tumors. J Clin Endocrinol Metab, 1998, 83: 4314-4320.
4 Kim K, Arai K, Sanno N, et al. Ghrelin and growth hormone (GH) secretagogue receptor (GHSR) mRNA expression in human pituitary adenomas. Clin Endocrinol (Oxf), 2001, 54: 759-768.
5雷霆,刘勤,刘暌,等.生长激素第三调节途径在人垂体生长激素腺瘤生长激素释放中的作用.中华实验外科杂志, 1999, 16: 429-431.
6 Lei T, Adams EF, Buchfelder M, et al. Relationship between protein kinase C and adenylyl cyclase activity in the regulation of growth hormone secretion by human pituitary somatotrophinomas. Neurosurgery, 1996, 39: 569-575.
7 Matsuzaki Y, Miyazawa K, Yokota T, et al. Molecular cloning and characterization of the human p19 (INK4d) gene promoter. FEBS Lett, 2002, 517: 272-276.
8 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
9冯作化,主编.医学分子生物学.第一版.北京:人民卫生出版社,2001: 61.
10 Alam J, Cook JL. Reporter genes: application to the study of mammalian gene transcription. Anal Biochem, 1999, 188(2): 245-254.
11 Suto CM, Ignar DM. Selection of an optimal reporter gene for cell-based high throughput screening assays. J Biomol Screen, 1997, 2:7-9.
12刘志锋,姜勇.报告基因技术的理论基础及其应用.生理科学进展, 2002, 33(4): 361-363.
13 Eyck P, Sabine B, Ulrich L, et al. Molecular cloing of a functional promoter of thehuman plakoglobin gene. Eur J Endocrinol, 2001, 145(2): 625-633.
14 Vasudevan N, Bahadur U, Kondaiah P. Characterization of chicken riboflavin carrier protein gene structure and promoter regulation by estrogen. J Biosci, 2001, 26(1): 39-46.
1 Antonson P, Al-Beidh F, Matthews J, et al. The human RAP250gene: genomic structure and promoter analysis. Gene, 2004, 327(2): 233-238.
2 Kobori H, Hayshi M, Saruta T. Throid Hormone Stimulates Renin Gene Expression Through the Thyroid Hormone Response Element. Hypertension, 2001, 37(1): 99-104.
3 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
4 Kaji H, Tai S, Okimura Y, et al. Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. J Biol Chem, 1998, 273: 85-88.
5冯作化,主编.医学分子生物学.第一版.北京:人民卫生出版社, 2001: 61.
6巨立中,成军,钟彦伟.启动子DNA结合蛋白研究策略.世界华人消化杂志, 2004, 12: 141-142.
7萨姆布鲁克J,拉塞尔DW.分子克隆实验指南.黄培堂译.第3版.北京:科学出版社,2002: 1347-1350.
8刘志峰,姜勇.报告基因技术的理论基础及其应用.生理科学进展, 2002, 33(4): 361-363.
9 Alam J, Cook JL. Reporter genes: application to the study of mammalian gene transcription. Anal Biochem, 1990, 188(2): 245-254.
10薛丽香,童坦军,张宗玉.报告基因的选择及其研究趋向.生理科学进展, 2002, 33(4): 364-366.
11 Eyck P, Sabine B, Ulrich L, et al. Molecular cloning of a functional promoter of the human plakoglobin gene. Eur J Endocrinol, 2001, 145(2): 625-633.
12 Vasudevan N, Bahadur U, Kondaiah P. Characterization of chicken riboflavin carrier protein gene structure and promoter regulation by estrogen. J Biosci. 2001, 26(1): 39-46.
13 Korbonits M, Jacobs RA, Aylwin SJB et al. Expression of the growth hormone secretagogue receptor in pituitary adenomas and other neuroendocrine tumors. J Clin Endoctinol Metab, 1998, 83: 3624-3630.
14 Poxon SW, Hughes JA. A biofuctional assay to study pRL-CMV plasmid DNA formulation stability. PDA J Pharm Sci Technol, 1999, 53(6): 314-317.
1 Yan M, Jones ME, Hernandez M, et al. Oestrogen replacement in vivo rescues the dysfunction of pituitary somatotropes in ovariectomised aromatase knockout mice. Neuroendocrinology, 2005, 81: 158-166.
2 Chen C. Growth hormone secretagogue actions on the pituitary gland: multiple receptors for multiple ligands? Clin Exp Pharmacol Physiol, 2000, 27: 323-329.
3 Katayama M, Nogami H, Nishiyama J, et al. Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland. Neuroendocrinology, 2000, 72: 333-340.
4 Kamegai J, Wakabayashi I, Miyamoto K, et al. Growth hormone-dependent regulation of pituitary GH secretagogue receptor (GHS-R) mRNA levels in the spontaneous dwarf Rat. Neuroendocrinology, 1998, 68: 312-318.
5 Petersenn S. Structure and regulation of the growth hormone secretagogue receptor. Minerva Endocrinol, 2002, 27: 243-256.
6 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001,142: 2649-2659.
7 Kaji H, Tai S, Okimura Y, et al. Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. J Biol Chem, 1998, 273: 85-88.
8 Nogueiras R, Tovar S, Mitchell SE, et al. Regulation of growth hormone secretagogue receptor gene expression in the arcuate nuclei of the rat by leptin and ghrelin. Diabetes, 2004, 53: 52-58.
9 Kim K, Arai K, Sanno N, et al. Ghrelin and growth hormone (GH) secretagogue receptor (GHSR) mRNA expression in human pituitary adenomas. Clin Endocrinol (Oxf), 2001, 54: 759-768.
10 Frutos MG, Cacicedo L, Fernandez C, et al. Insights into a role of GH secretagogues in reversing the age-related decline in the GH/IGF-I axis. Am J Physiol Endocrinol Metab, 2007, 293: 1140-1152.
11 Mano-Otagiri A, Nemoto T, Sekino A, et al. Growth hormone-releasing hormone (GHRH) neurons in the arcuate nucleus (Arc) of the hypothalamus are decreased in transgenic rats whose expression of ghrelin receptor is attenuated: Evidence that ghrelin receptor is involved in the up-regulation of GHRH expression in the arc. Endocrinology, 2006, 147: 4093-4103.
12 Luque RM, Peinado JR, Gracia-Navarro F, et al. Cortistatin mimics somatostatin by inducing a dual, dose-dependent stimulatory and inhibitory effect on growth hormone secretion in somatotropes. J Mol Endocrinol, 2006, 36: 547-556.
13 Gauna C, Van der Lely AJ. Somatostatin, cortistatin, ghrelin and glucose metabolism. J Endocrinol Invest, 2005, 28: 127-131.
14 Petersenn S, Rasch AC, Presch S, et al. Characterization of the human somatostatin receptor type 4 promoter. Mol Cell Endocrinol, 2002, 188: 75-83.
15 Sato M, Nakahara K, Miyazato M, et al. Regulation of GH secretagogue receptor gene expression in the rat nodose ganglion. J Endocrinol, 2007, 194: 41-46.
16 Kineman RD, Kamegai J, Frohman LA. Growth hormone (GH)-releasing hormone (GHRH) and the GH secretagogue (GHS), L692, 585, differentially modulate rat pituitary GHS receptor and GHRH receptor messenger ribonucleic acid levels. Endocrinology, 1999, 140: 3581-3586.
17 Bennett PA, Thomas GB, Howard AD, et al. Hypothalamic growth hormone secretagogue-receptor (GHS-R) expression is regulated by growth hormone in the rat. Endocrinology, 1997, 138: 4552-4557.
18 Mazzocchi G, Neri G, Rucinski M, et al. Ghrelin enhances the growth of cultured human adrenal zona glomerulosa cells by exerting MAPK-mediated proliferogenic and antiapoptotic effects. Peptides, 2004, 25: 1269-1277.
19雷霆,刘勤,刘睽等.生长激素第三调节途径在人垂体生长激素腺瘤生长激素释放中的作用.中华实验外科杂志, 1999, 16(5): 429-431.
20 Avramidis A, Polyzos SA, Efstathiadou Z, et al. Sustained Clinical Inactivity and Stabilization of GH/IGF-1 Levels in an Acromegalic Patient after Discontinuation of Somatostatin Analogue Treatment. Endocr J, 2008, 31: 113-117.
21 Martinez-Delgado IA, Gomez-Martinez G. [Usefulness of GH and IGF-1 to establishthe dose and frequency of application of octreotide to treat acromegaly]. Rev Med Inst Mex Seguro Soc, 2007, 45: 291-295.
22 Maiza JC, Vezzosi D, Matta M, et al. Long-term (up to 18 years) effects on GH/IGF-1 hypersecretion and tumour size of primary somatostatin analogue (SSTa) therapy in patients with GH-secreting pituitary adenoma responsive to SSTa. Clin Endocrinol (Oxf), 2007, 67: 282-289.
23 Ghigo E, Gianotti L, Arvat E, et al. Effects of recombinant human insulin-like growth factor I administration on growth hormone (GH) secretion, both spontaneous and stimulated by GH-releasing hormone or hexarelin, a peptidyl GH secretagogue, in humans. J Clin Endocrinol Metab, 1999, 84: 285-290.
24 Thomas GB, Fairhall KM, Robinson IC. Activation of the hypothalamo-pituitary-adrenal axis by the growth hormone (GH) secretagogue, GH-releasing peptide-6, in rats. Endocrinology, 1997, 138: 1585-1591.
25 Tamura H, Kamegai J, Sugihara H, et al. Glucocorticoids regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2000, 12: 481-485.
26 Nagaki S, Fukamauchi F, Sakamoto Y, et al. Upregulation of brain somatostatin and neuropeptide Y following lidocaine-induced kindling in the rat. Brain Res, 2000, 852: 470-474.
27 Kamegai J, Tamura H, Ishii S, et al. Thyroid hormones regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2001, 13: 275-278.
28 Balasubramanian G, Hanumegowda U, Reddy CS. Secalonic acid D alters the nature of and inhibits the binding of the transcription factors to the phorbol
12-O-tetradecanoate-13 acetate-response element in the developing murine secondary palate. Toxicol Appl Pharmacol, 2000, 169: 142-150.
29 Kamegai J, Wakabayashi I, Kineman RD, et al. Growth hormone-releasing hormone receptor (GHRH-R) and growth hormone secretagogue receptor (GHS-R) mRNA levels during postnatal development in male and female rats. J Neuroendocrinol, 1999, 11: 299-306.
30 Kamegai J, Tamura H, Shimizu T, et al. Estrogen receptor (ER) alpha, but not ERbeta, gene is expressed in growth hormone-releasing hormone neurons of the male rat hypothalamus. Endocrinology, 2001, 142: 538-543.
1 Dominguez B, Avila T, Flores-Hernandez J, et al. Up-regulation of High Voltage-activated Ca (2+) Channels in GC Somatotropes after Long-term Exposure to Ghrelin and Growth Hormone Releasing Peptide-6. Cell Mol Neurobiol, 2008, 1: 77-81.
2 Pong S, Chaung LP, Dean CD, et al. Identification of new G-protein-linked receptor for growth hormone secretagogues. Mol Endocrinol, 2004, 10: 57-61.
3 Howard AD, Feighner SD, Cully DF, et al. A receptor in pituitary and hypothalamus that functions in growth hormone release.Science, 2002, 273: 974-977.
4 McKee KK, Palyha OC, Feighner SD, et al. Molecular analysis of rat pituitary and hypothalamic growth hormone secretagogue receptors. Mol Endocrinol, 1997, 11: 415-423.
5 Kaji H, Tai S, Okimura Y, et al. Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. 78th Annual Meeting of the Endocrine Society, New Orleans, 1998: 33-40.
6 Guan X, Yu H, Palyha OC, et aI. Distribution of mRNA encoding the growth hormone secretagogue receptor in brain and peripheral tissues. Mol Brain Res, 1998, 48: 23-29.
7 Bennett PA, Thomas GB, Howard AD, et al. Hypothalamic growth hormone secretagogue-receptor (GHS-R) expression is regulated by growth hormone in the rat. Endocrinology, 1997, 138: 4552-4557.
8 Yokote R, Sato M, Matsubara S, et al. Molecular cloning and gene expression of growth hormone-releasing peptide receptor in rat tissues. Peptides, 1998, 19: 15-20.
9 Tarmenbaum GS, Lapointe M, Beandet A, et al. Expression of growth hormone secretagogue receptors by growth hormone-releasing hormone neurons in the mediobasal hypothalamus. Endocrinology, 1998, 139: 4420-4423.
10 Hreniuk DL, Palyha OC, Howard AD, et al. Immunocytochemical localisation of a growth hormone secretagogue receptor in rat pituitary cells. 78th Annual Meeting of the Endocrine Society, New Orleans, 1998: 220-224.
11 Ong H, Bodfot V, McNicoll N, et al. Identification and characterization of a newGHRP receptor in the heart. GH IGF Res, 1998, 8: 137-140.
12 Locatelli V, Rossoni G, DeGennaro Colonna V, et al. Evidence for direct cardiac effects of hexarelin in the hypophysectomised rat. 78th Annual Meeting of the Endocrine Society, New Orleans, 1998: P2-247.
13 Feighner SD, Howard AID, Prendergast K, et al. Structural requirements for the activation of the human growth hormone secretagogue receptor by peptide and nonpeptide secretagogues. Mol Endocrinol, 1998, 12: 137-145.
14 Smith RG, Van der Ploeg LHT, Howard AD, et al. Peptidomimetic regulation of growth hormone secretion. Endocr Rev, 1997, 18: 621-645.
15 Nargund RP, Van der Ploeg LHT. Growth hormone secretagogues. In: Bristol JA, ed. Annual reports in medicinal chemistry, VoI. 32. San Diego: Academic Press, 1997: 221-230.
16 McKee KK, Tan CP, Palyha OC, et al. Cloning and characterization of two human G protein-coupled receptor genes (GPR38 and GPR39) related to the growth hormone secretagogue and neurotensin receptors. Genomics, 1997, 46: 426-434.
17 Ong H, McNicoll N, Escher E, et al. Identification of a pituitary growth hormone-releasing peptide (GHRP) receptor subtype by photoaffinity labeling. Endocrinology, 1998, 139: 432-435.
18 Muccioli G, Ghe C, Ghigo MC, et al. Specific receptors for synthetic GH secretagogues in the human brain and pituitary gIand. J Endocrinol, 1998, 157: 99-106.
19 De Gennaro Colonna V, Rossoni G, Bernareggi M, et al. Cardiac ischemia and impairment of vascular endothelinm function in hearts from growth hormone-deficient rats: protection by hexarelin. Eur J Pharmacol, 1997, 334: 201-207.
20 Tan CE, McKee KK, Lin O, et al. Cloning and characterisation of a human and murine T-cell orphan G-protein-coupled receptor similar to the growth hormone secretagogue and neurotensin receptors. Genomics, 1998, 52: 223-229.
21 Cheng K, Chan W-W, Barreto A, et al. The synergistic effects of His-D-Trp-Ala- Trp-D-Phe-Lys-NH2 on growth hormone (GH)-releasing factor-stimulated GH release and intracellular adenosin 3', 5’-monophosphate accumulation in rat pituitary cell culture. Endocrinology, 1989, 124: 2791-2798.
22 Cheng K, Chan WW, Butler B, et al. Evidence for a role of protein kinase-C in His-D-Trp-Ala-Trp-D-Phe- Lys-NH2-induced growth hormone release from rat primary pituitary cells. Endocrinology, 1991, 129: 3337-3342.
23 Akman MS, Girard M, O'Brien LE Ho AK, et al. Mechanisms of action of a second generation growth hormone-releasing peptide (Ala-His-D-beta-Nal-Ala-Trp- D-Phe-Lys-NH2) in rat anterior pituitary cells. Endocrinology, 1993, 132: 1286-1291.
24 Pong SS, Chaung LY, Smith RG. GHRP-6 (His-D-Trp-Ala-Trp- D-Phe-Lys-NH2) stimulates growth hormone secretion by depolarization in rat pituitary cell cultures.
73rd Annual Meeting of the Endocrine Society, Washington, 1991.
25 Herrington J, Hille B. Growth hormone-releasing hexapeptide elevates intracellular calcium in rat somatotropes by 2 mechanisms. Endocrinology, 1994, 135: 1100-1108.
26 Wu D, Chen C, Katoh K, et al. The effect of GH-releasingpeptide- 2 (GHRP-2 or KP 102) on GH secretion from primary cultured ovine pituitary ceils can be abolished by a specific GH-releasing factor (GRF) receptor antagonist. J Endocrinol, 1994, 140: R8-R13.
27 Cheng K, Chart WW, Butler B, et al. Stimulation of growth hormone release from rat primary pituitary cells by L-692,429, a novel non-peptidyl GH secretagogue. Endocrinology, 1993, 132: 2729-2731.
28 Adams EE, Lei T, Bucherfelder M, et al. Protein kinase C-dependent growth hormone releasing peptides stimulate cyclic adenosine 3', 5’-monophosphate production by human pituitary somatotropinomas expressing gsp oncogenes: evidence for cross-talk between transduction pathways. Mol Endocrinol, 1996, 10: 432-438.
29 Bresson-Bepoldin L, Odessa MF, Dufy-Barbe L. GHRP-6 stimulates calcium increase and growth hormone release in human somatotrophs in vivo. Endocrine, 1994, 2: 793-803.
30 Adams EE, Lei T, Buchfelder M, et al. Biochemical characteristics of human pituitary somatotrophinomas with or without gsp mutations. J Clin Endocrinol Metab, 1995, 80: 2077-2081.
31 Adams EF, Buchfelder M, Lei T, et al. In vitro responses of GH-secreting tumours with and without gsp oncogenes to octreotide, GHRH and growth hormone-releasingpeptide. In: von Werder K, Fahlbusch Reds. Pituitary adenomas: from basic research to diagnosis and therapy. Amsterdam: Elsevier, 1996: 43-47.
32 Korborins M, Adams EF, Grossman AB. The effects of GHsecretagognes on human pituitary cells in culture and on rat hypothalamic tissue. In: Dieguez C, Ghigo E, Boghen M, Casanueva FF, eds. Growth hormone secretagogues: basic findings and clinical implications. Rotterdam: Elsevier, 1998 (in press).
33 Lei T, Bucherfelder M, Fahlbush R, et al. Growth hormonereleasing peptide (GHRP-6) stimulates phosphatidylinositol (PI) turnover in human pituitary somatotroph cells. J Mol Endocrinol, 1995, 14: 135-138.
34 Houslay MD. Crosstalk: a pivotal rote for the protein kinase C in modulating relationships between signal transduction pathways. J Biol Chem, 1998, 264: 8802-8810.
35 Cheng J, Wu TJ, Butler B, et al. Growth hormone releasing peptides: a comparison of the growth hormone releasing activities of GHRP-2 and GHRP-6 in rat primary pituitary cells. Life Sci, 1997, 60: 1385-1392.
36 Hickey GJ, Jacks TM, Schleim KD, et al. Repeat administration of the GH secretagogue MK-0677 increases and maintains elevated IGE-I levels in beagles. J Endocrinol, 1997, 152: 183-192.
37 Carmignac DF, Bennett PA, Robinson ICAE. Effects of growth hormone secretagogues on prolactin release in anesthetized dwarf (dw/dw) rats. Endocrinology, 1998, 139: 3590-3596.
38 Arvat E, Gianotti L, Broglio F, et al. Oestrogen replacement does not restore the reduced GH-releasing activity of hexarelin, a synthetic hexapeptide, in post-menopausal women. Eur J Endocrinol, 1997, 136: 483-487.
39 Korbonits M, Jacobs RA, Aylwin S, et al. Expression of the growth hormone secretagogue receptor in pituitary adenomas and other neuroendocrine tumors. J Clin Endoctinol Metab, 1998, 83: 3624-3630.
40 Haddad G, Penabad JL, Bashey HM, et al. Expression of activin/inhibin subunit messenger ribonucleic acids by gonadotroph adenomas. J Clin Endocrinol Metab, 1994, 79: 1399-1403.
41 Renner U, Brockmeier S, Strasburger CJ, et al. Growth-hormone (GH)-releasing peptide stimulation of GH release from human somatotroph adenoma cells - interaction with GH-releasing hormone, thyrotropin-releasing-hormone, and octreotide. J Clin Endocrinol Metab, 1994, 78: 1090-1096.
42 Dekeyzer Y, Lenne E, Bertagna X. Widespread transcription of the growth hormone-releasing peptide receptor gene in neuroendocfine human tumors. Eur J Endocrinol, 1997, 137: 715-718.
43 Lania A, Ballar6 E, Corbetta S, et al. Growth hormone-releasing peptide (GHRP-6) increases intracellular calcium concentrations in cultured cells from human pituitary adenomas of different types. Eur J Endocrinol, 1998, 139: 343-348.
44 Adams EF, Huang B, Buchfelder M, et al. Presence of growth hormone secretagogue receptor messenger ribonucleic acid in human pituitary tumors and rat GH3 cells. J Clin EndocrinoI Metab, 1998, 83: 638-642.
45 Nielsen S, Mellemkjaer S, Rasmussen L, et al. Gene transcription of receptors for growth hormone-releasing peptide and somatostatin in human pituitary adenomas. J Clin Endocrinol Metab, 1998, 83: 2997-3000.
46 Barlier A, Grino M, Zamora AJ, et al. Expression of growth hormone secretagogue-receptors (GHS-R) in human pituitary adenomas. European Congress of Endocrinology, Seville, 1998: 332-341.
47 Jansson J, Svensson J, Bengtsson B, et al. Acromegaly and Cushing's syndrome due to ectopic production of GHRH and ACTH by a thymic carcinoid tumour: in vitro responses to GHRH and GHRP-6. Clin Endocrinol, 1998, 48: 243-250.
48 Hosoda H, Kojima M, Matsuo H, et al. Purification and characterization of rat des-Gln14-Ghrelin, a second endogenous ligand for the growth hormone secretagogue receptor. J Biol Chem, 2000, 275(29): 21995-22000.
49 Date Y, Kojima M, Hosoda H, et al. Ghrelin, a novel growth hormone-releasing acylated peptide, is synthesized in a distinct endocrine cell type in the gastrointestinal tracts of rats and humans. Endocrinology, 2000, 141(11): 4255-4261.
50 Nakazato M, Murakami N, Date Y, et al. A role for ghrelin in the central regulation of feeding. Nature, 2001, 409(6817): 194-198.
51 Nagaya N, Kojima M, Uematsu M, et al. Hemodynamic and hormonal effects of human ghrelin in healthy volunteers. Am J Physiol Regul Integr Comp Physiol, 2001, 280(5): 1483-1487.
52 Date Y, Murakami N, Kojima M, et al. Central effects of a novel acylated peptide, ghrelin on growth hormone release in rats. Biochem Biophys Res Commun, 2000, 275(2): 477-480.
53 Wren AM, Small CJ, Ward HL, et al. The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion. Endocrinology, 2000, 141(11): 4325-4328.
54 Shintani M, Ogawa Y, Ebihara K, et al. Ghrelin, an endogenous growth hormone secretagogue, is a novel orexigenic peptide that antagonizes leptin action through the activation of hypothalamic neuropeptide Y/Y1 receptor pathway. Diabetes, 2001, 50(2): 227-232.
55 Kojima M, Hosodo H, Date Y, et al. Ghrelin is a growth hormone releasing acylated peptide from stomach. Nature, 1999, 402(6762): 656-660.
56 Banks WA, Tschop M, Robinson SM, et al. Extent and direction of ghrelin transport across the blood–brain barrier is determined by its unique primary structure. J Pharmacol Exp Ther, 2002, 302(2): 822-827.
57 Bednarek MA, Feighner SD, Pong SS, et al. Structure–function studies on the new growth hormone-releasing peptide, ghrelin: minimal sequence of ghrelin necessary for activation of growth hormone secretagogue receptor 1a. J Med Chem, 2000, 43: 4370- 4376.
58 Muccioli G, Papotti M, Locatelli V, et al. Binding of 125I-labeled ghrelin to membranes from human hypothalamus and pituitary gland. J Endocrin Invest, 2001, 24: 7-9.
59 Date Y, Kojima M, Hosoda H, et al. Ghrelin, a novel growth hormone-releasing acylated peptide, is synthesised in a distinct endocrine cell type in the gastrointestinal tract of rats and humans. Endocrinology, 2000, 141: 4255-4261.
60 Seoane LM, Tovar S, Baldelli R, et al. Ghrelin elicits a marked stimulatory effect on GH secretion in freely-moving rats. Eur J Endocrinol, 2000, 143: 7-9.
61 Arvat E, Di Vito L, Broglio M, et al. Preliminary evidence that ghrelin, the natural GH secretagogue (GHS)-receptor ligand, strongly stimulates GH secretion in humans. J Endocrin Invest, 2000, 23: 493-495.
62 Peino R, Baldelli R, Rodriguez-Garcia J, et al. Ghrelin-induced growth hormone secretion in humans. Eur J Endocrinol, 2000, 143: 11-14.
63 Takaya K, Ariyasu H, Kanamoto N, et al. Ghrelin strongly stimulates growth hormone release in humans. J Clin Endocrinol Metab, 2000, 85: 4908-4911.
64 Kamegai J, Tamura H, Ishii S, Sugihara H, et al. Thyroid hormones regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2001, 13: 275-278.
65 Tamura H, Kamegai J, Sugihara H, et al. Glucocorticoids regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2000, 12: 481- 485.
66 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
67 Katayama M, Nogami H, Nishiyama J, et al. Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland. Neuroendocrinology, 2000, 72: 333-340.
68 Barlier A, Zamora AJ, Grino M, et al. Expression of functional growth hormone secretagogue receptors in human pituitary adenomas: polymerase chain reaction, triple in situ hybridization and cell culture studies. J Neuroendocrinol, 1999, 11: 491-502.
69 Smith RG, Feighner S, Prendergast K, et al. A new orphan receptor involved in pulsatile growth hormone release. Trends Endocrinol Metab, 1999, 10(4): 128-135.
70 Cheng K, Chan WWS, Butler B, et al. Evidence for a role of protein kinase-C in His-D-Trp-Ala-D-Phe-Lys-amide induced growth hormone release from rat pituitary cells. Endocrinology, 1991, 129: 3337-3342
71 Luton J, Cerdas S, Billaud L, et al. Clinical features of adrenocortical carcinoma, prognostic factors, and the effect of mitotane therapy. N Engl J Med, 1990, 322: 1195-1201.
72 Mazzocchi G, Neri G, Rucinski M, et al. Ghrelin enhances the growth of cultured human adrenal zona glomerulosa cells by exerting MAPK-mediated proliferogenic and antiapoptotic effects. Peptides, 2004, 25: 1269-1277.
73 Muccioli G, Pons N, Ghe C, et al. Ghrelin and des-acyl ghrelin both inhibit isoproterenol-induced lipolysis in rat adipocytes via a non-type 1a growth hormone secretagogue receptor. Eur J Pharmacol, 2004, 498: 27-35.
74 Nanzer AM, Khalaf S, Mozid AM, et al. Ghrelin exerts a proliferative effect on a rat pituitary somatotroph cell line via the mitogen-activated protein kinase pathway. Eur J Endocrinol, 2004, 151: 233-240.
75 Nogueiras R, Tovar S, Mitchell SE, et al. Regulation of growth hormone secretagogue receptor gene expression in the arcuate nuclei of the rat by leptin and ghrelin. Diabetes, 2004, 53: 2552-2558.
76 Thompson NM, Gill DA, Davies R, et al. Ghrelin and des-octanoyl ghrelin promote adipogenesis directly in vivo by a mechanism independent of GHS-R1a. Endocrinology, 2003, 145: 234-242.
77 Tortorella C, Macchi C, Spinazzi R, et al. Ghrelin, an endogenous ligand for the growth hormonesecretagogue receptor, is expressed in the human adrenal cortex. Int J Mol Med,2003, 12: 213-217.
2 Pong S, Chaung LP, Dean CD, et al. Identification of a new G-protein-link receptor for growth hormone secretagogues. Mol Endocrinol, 1996, 10: 57-61.
3 Howard AD, Feighner SD, Cully DF, et al. A receptor in pituitary and hypothalamus that functions in growth hormone release. Science, 1996, 273: 974-977.
4 Simith RG. Development of growth hormone secretagogues. Endocr Rev, 2005, 26 (3): 346-360.
5 Kamegai J, Tamura H, Shimizu T, et al. The role of pituitary ghrelin in growth hormone (GH) secretion: GH-releasinghormone-dependent regulation of pituitary ghrelin gene expression and peptide content. Endocrinology, 2004, 145(8): 3731-3738.
6 Volante M, Allia E, Gugliotta P, et al. Expression of ghrelin and of the GH secretagogue receptor by pancreatic islet cells and related endocrine tumors. J Clin Endocrinol Metab, 2002, 87: 1300-1308.
7 Penny L, Adrian C, Lisa K. The potential autocrine/paracrine roles of ghrelin and its receptor in hormone-dependent cancer. Cytokine & Growth Factor Reviews, 2003, 14: 113-122.
8 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
9 Yan M, Jones ME, Hernandez M, et al. Oestrogen replacement in vivo rescues the dysfunction of pituitary somatotropes in ovariectomised aromatase knockout mice. Neuroendocrinology, 2005, 81: 158-166.
10 Chen C. Growth hormone secretagogue actions on the pituitary gland: multiple receptors for multiple ligands? Clin Exp Pharmacol Physiol, 2000, 27: 323-329.
11 Katayama M, Nogami H, Nishiyama J, et al. Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland. Neuroendocrinology, 2000, 72: 333-340.
12 Kamegai J, Wakabayashi I, Miyamoto K, et al. Growth hormone-dependent regulation of pituitary GH secretagogue receptor (GHS-R) mRNA levels in the spontaneous dwarf Rat. Neuroendocrinology, 1998, 68: 312-318.
1 Bertherat J. Cloning of the growth hormone secretagogues receptor cDNA: new evidence for a third endocrine pathway controlling growth hormone release. Eur J Endocrinol, 1997, 136: 37-38.
2 Smith RG. Development of growth hormone secretagogues. Endocr Rev, 2005, 26: 346-360.
3 Skinner MM, Nass R, Lopes B, et al. Growth hormone secretagogue receptor expression in human pituitary tumors. J Clin Endocrinol Metab, 1998, 83: 4314-4320.
4 Kim K, Arai K, Sanno N, et al. Ghrelin and growth hormone (GH) secretagogue receptor (GHSR) mRNA expression in human pituitary adenomas. Clin Endocrinol (Oxf), 2001, 54: 759-768.
5雷霆,刘勤,刘暌,等.生长激素第三调节途径在人垂体生长激素腺瘤生长激素释放中的作用.中华实验外科杂志, 1999, 16: 429-431.
6 Lei T, Adams EF, Buchfelder M, et al. Relationship between protein kinase C and adenylyl cyclase activity in the regulation of growth hormone secretion by human pituitary somatotrophinomas. Neurosurgery, 1996, 39: 569-575.
7 Matsuzaki Y, Miyazawa K, Yokota T, et al. Molecular cloning and characterization of the human p19 (INK4d) gene promoter. FEBS Lett, 2002, 517: 272-276.
8 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
9冯作化,主编.医学分子生物学.第一版.北京:人民卫生出版社,2001: 61.
10 Alam J, Cook JL. Reporter genes: application to the study of mammalian gene transcription. Anal Biochem, 1999, 188(2): 245-254.
11 Suto CM, Ignar DM. Selection of an optimal reporter gene for cell-based high throughput screening assays. J Biomol Screen, 1997, 2:7-9.
12刘志锋,姜勇.报告基因技术的理论基础及其应用.生理科学进展, 2002, 33(4): 361-363.
13 Eyck P, Sabine B, Ulrich L, et al. Molecular cloing of a functional promoter of thehuman plakoglobin gene. Eur J Endocrinol, 2001, 145(2): 625-633.
14 Vasudevan N, Bahadur U, Kondaiah P. Characterization of chicken riboflavin carrier protein gene structure and promoter regulation by estrogen. J Biosci, 2001, 26(1): 39-46.
1 Antonson P, Al-Beidh F, Matthews J, et al. The human RAP250gene: genomic structure and promoter analysis. Gene, 2004, 327(2): 233-238.
2 Kobori H, Hayshi M, Saruta T. Throid Hormone Stimulates Renin Gene Expression Through the Thyroid Hormone Response Element. Hypertension, 2001, 37(1): 99-104.
3 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
4 Kaji H, Tai S, Okimura Y, et al. Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. J Biol Chem, 1998, 273: 85-88.
5冯作化,主编.医学分子生物学.第一版.北京:人民卫生出版社, 2001: 61.
6巨立中,成军,钟彦伟.启动子DNA结合蛋白研究策略.世界华人消化杂志, 2004, 12: 141-142.
7萨姆布鲁克J,拉塞尔DW.分子克隆实验指南.黄培堂译.第3版.北京:科学出版社,2002: 1347-1350.
8刘志峰,姜勇.报告基因技术的理论基础及其应用.生理科学进展, 2002, 33(4): 361-363.
9 Alam J, Cook JL. Reporter genes: application to the study of mammalian gene transcription. Anal Biochem, 1990, 188(2): 245-254.
10薛丽香,童坦军,张宗玉.报告基因的选择及其研究趋向.生理科学进展, 2002, 33(4): 364-366.
11 Eyck P, Sabine B, Ulrich L, et al. Molecular cloning of a functional promoter of the human plakoglobin gene. Eur J Endocrinol, 2001, 145(2): 625-633.
12 Vasudevan N, Bahadur U, Kondaiah P. Characterization of chicken riboflavin carrier protein gene structure and promoter regulation by estrogen. J Biosci. 2001, 26(1): 39-46.
13 Korbonits M, Jacobs RA, Aylwin SJB et al. Expression of the growth hormone secretagogue receptor in pituitary adenomas and other neuroendocrine tumors. J Clin Endoctinol Metab, 1998, 83: 3624-3630.
14 Poxon SW, Hughes JA. A biofuctional assay to study pRL-CMV plasmid DNA formulation stability. PDA J Pharm Sci Technol, 1999, 53(6): 314-317.
1 Yan M, Jones ME, Hernandez M, et al. Oestrogen replacement in vivo rescues the dysfunction of pituitary somatotropes in ovariectomised aromatase knockout mice. Neuroendocrinology, 2005, 81: 158-166.
2 Chen C. Growth hormone secretagogue actions on the pituitary gland: multiple receptors for multiple ligands? Clin Exp Pharmacol Physiol, 2000, 27: 323-329.
3 Katayama M, Nogami H, Nishiyama J, et al. Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland. Neuroendocrinology, 2000, 72: 333-340.
4 Kamegai J, Wakabayashi I, Miyamoto K, et al. Growth hormone-dependent regulation of pituitary GH secretagogue receptor (GHS-R) mRNA levels in the spontaneous dwarf Rat. Neuroendocrinology, 1998, 68: 312-318.
5 Petersenn S. Structure and regulation of the growth hormone secretagogue receptor. Minerva Endocrinol, 2002, 27: 243-256.
6 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001,142: 2649-2659.
7 Kaji H, Tai S, Okimura Y, et al. Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. J Biol Chem, 1998, 273: 85-88.
8 Nogueiras R, Tovar S, Mitchell SE, et al. Regulation of growth hormone secretagogue receptor gene expression in the arcuate nuclei of the rat by leptin and ghrelin. Diabetes, 2004, 53: 52-58.
9 Kim K, Arai K, Sanno N, et al. Ghrelin and growth hormone (GH) secretagogue receptor (GHSR) mRNA expression in human pituitary adenomas. Clin Endocrinol (Oxf), 2001, 54: 759-768.
10 Frutos MG, Cacicedo L, Fernandez C, et al. Insights into a role of GH secretagogues in reversing the age-related decline in the GH/IGF-I axis. Am J Physiol Endocrinol Metab, 2007, 293: 1140-1152.
11 Mano-Otagiri A, Nemoto T, Sekino A, et al. Growth hormone-releasing hormone (GHRH) neurons in the arcuate nucleus (Arc) of the hypothalamus are decreased in transgenic rats whose expression of ghrelin receptor is attenuated: Evidence that ghrelin receptor is involved in the up-regulation of GHRH expression in the arc. Endocrinology, 2006, 147: 4093-4103.
12 Luque RM, Peinado JR, Gracia-Navarro F, et al. Cortistatin mimics somatostatin by inducing a dual, dose-dependent stimulatory and inhibitory effect on growth hormone secretion in somatotropes. J Mol Endocrinol, 2006, 36: 547-556.
13 Gauna C, Van der Lely AJ. Somatostatin, cortistatin, ghrelin and glucose metabolism. J Endocrinol Invest, 2005, 28: 127-131.
14 Petersenn S, Rasch AC, Presch S, et al. Characterization of the human somatostatin receptor type 4 promoter. Mol Cell Endocrinol, 2002, 188: 75-83.
15 Sato M, Nakahara K, Miyazato M, et al. Regulation of GH secretagogue receptor gene expression in the rat nodose ganglion. J Endocrinol, 2007, 194: 41-46.
16 Kineman RD, Kamegai J, Frohman LA. Growth hormone (GH)-releasing hormone (GHRH) and the GH secretagogue (GHS), L692, 585, differentially modulate rat pituitary GHS receptor and GHRH receptor messenger ribonucleic acid levels. Endocrinology, 1999, 140: 3581-3586.
17 Bennett PA, Thomas GB, Howard AD, et al. Hypothalamic growth hormone secretagogue-receptor (GHS-R) expression is regulated by growth hormone in the rat. Endocrinology, 1997, 138: 4552-4557.
18 Mazzocchi G, Neri G, Rucinski M, et al. Ghrelin enhances the growth of cultured human adrenal zona glomerulosa cells by exerting MAPK-mediated proliferogenic and antiapoptotic effects. Peptides, 2004, 25: 1269-1277.
19雷霆,刘勤,刘睽等.生长激素第三调节途径在人垂体生长激素腺瘤生长激素释放中的作用.中华实验外科杂志, 1999, 16(5): 429-431.
20 Avramidis A, Polyzos SA, Efstathiadou Z, et al. Sustained Clinical Inactivity and Stabilization of GH/IGF-1 Levels in an Acromegalic Patient after Discontinuation of Somatostatin Analogue Treatment. Endocr J, 2008, 31: 113-117.
21 Martinez-Delgado IA, Gomez-Martinez G. [Usefulness of GH and IGF-1 to establishthe dose and frequency of application of octreotide to treat acromegaly]. Rev Med Inst Mex Seguro Soc, 2007, 45: 291-295.
22 Maiza JC, Vezzosi D, Matta M, et al. Long-term (up to 18 years) effects on GH/IGF-1 hypersecretion and tumour size of primary somatostatin analogue (SSTa) therapy in patients with GH-secreting pituitary adenoma responsive to SSTa. Clin Endocrinol (Oxf), 2007, 67: 282-289.
23 Ghigo E, Gianotti L, Arvat E, et al. Effects of recombinant human insulin-like growth factor I administration on growth hormone (GH) secretion, both spontaneous and stimulated by GH-releasing hormone or hexarelin, a peptidyl GH secretagogue, in humans. J Clin Endocrinol Metab, 1999, 84: 285-290.
24 Thomas GB, Fairhall KM, Robinson IC. Activation of the hypothalamo-pituitary-adrenal axis by the growth hormone (GH) secretagogue, GH-releasing peptide-6, in rats. Endocrinology, 1997, 138: 1585-1591.
25 Tamura H, Kamegai J, Sugihara H, et al. Glucocorticoids regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2000, 12: 481-485.
26 Nagaki S, Fukamauchi F, Sakamoto Y, et al. Upregulation of brain somatostatin and neuropeptide Y following lidocaine-induced kindling in the rat. Brain Res, 2000, 852: 470-474.
27 Kamegai J, Tamura H, Ishii S, et al. Thyroid hormones regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2001, 13: 275-278.
28 Balasubramanian G, Hanumegowda U, Reddy CS. Secalonic acid D alters the nature of and inhibits the binding of the transcription factors to the phorbol
12-O-tetradecanoate-13 acetate-response element in the developing murine secondary palate. Toxicol Appl Pharmacol, 2000, 169: 142-150.
29 Kamegai J, Wakabayashi I, Kineman RD, et al. Growth hormone-releasing hormone receptor (GHRH-R) and growth hormone secretagogue receptor (GHS-R) mRNA levels during postnatal development in male and female rats. J Neuroendocrinol, 1999, 11: 299-306.
30 Kamegai J, Tamura H, Shimizu T, et al. Estrogen receptor (ER) alpha, but not ERbeta, gene is expressed in growth hormone-releasing hormone neurons of the male rat hypothalamus. Endocrinology, 2001, 142: 538-543.
1 Dominguez B, Avila T, Flores-Hernandez J, et al. Up-regulation of High Voltage-activated Ca (2+) Channels in GC Somatotropes after Long-term Exposure to Ghrelin and Growth Hormone Releasing Peptide-6. Cell Mol Neurobiol, 2008, 1: 77-81.
2 Pong S, Chaung LP, Dean CD, et al. Identification of new G-protein-linked receptor for growth hormone secretagogues. Mol Endocrinol, 2004, 10: 57-61.
3 Howard AD, Feighner SD, Cully DF, et al. A receptor in pituitary and hypothalamus that functions in growth hormone release.Science, 2002, 273: 974-977.
4 McKee KK, Palyha OC, Feighner SD, et al. Molecular analysis of rat pituitary and hypothalamic growth hormone secretagogue receptors. Mol Endocrinol, 1997, 11: 415-423.
5 Kaji H, Tai S, Okimura Y, et al. Cloning and characterization of the 5'-flanking region of the human growth hormone secretagogue receptor gene. 78th Annual Meeting of the Endocrine Society, New Orleans, 1998: 33-40.
6 Guan X, Yu H, Palyha OC, et aI. Distribution of mRNA encoding the growth hormone secretagogue receptor in brain and peripheral tissues. Mol Brain Res, 1998, 48: 23-29.
7 Bennett PA, Thomas GB, Howard AD, et al. Hypothalamic growth hormone secretagogue-receptor (GHS-R) expression is regulated by growth hormone in the rat. Endocrinology, 1997, 138: 4552-4557.
8 Yokote R, Sato M, Matsubara S, et al. Molecular cloning and gene expression of growth hormone-releasing peptide receptor in rat tissues. Peptides, 1998, 19: 15-20.
9 Tarmenbaum GS, Lapointe M, Beandet A, et al. Expression of growth hormone secretagogue receptors by growth hormone-releasing hormone neurons in the mediobasal hypothalamus. Endocrinology, 1998, 139: 4420-4423.
10 Hreniuk DL, Palyha OC, Howard AD, et al. Immunocytochemical localisation of a growth hormone secretagogue receptor in rat pituitary cells. 78th Annual Meeting of the Endocrine Society, New Orleans, 1998: 220-224.
11 Ong H, Bodfot V, McNicoll N, et al. Identification and characterization of a newGHRP receptor in the heart. GH IGF Res, 1998, 8: 137-140.
12 Locatelli V, Rossoni G, DeGennaro Colonna V, et al. Evidence for direct cardiac effects of hexarelin in the hypophysectomised rat. 78th Annual Meeting of the Endocrine Society, New Orleans, 1998: P2-247.
13 Feighner SD, Howard AID, Prendergast K, et al. Structural requirements for the activation of the human growth hormone secretagogue receptor by peptide and nonpeptide secretagogues. Mol Endocrinol, 1998, 12: 137-145.
14 Smith RG, Van der Ploeg LHT, Howard AD, et al. Peptidomimetic regulation of growth hormone secretion. Endocr Rev, 1997, 18: 621-645.
15 Nargund RP, Van der Ploeg LHT. Growth hormone secretagogues. In: Bristol JA, ed. Annual reports in medicinal chemistry, VoI. 32. San Diego: Academic Press, 1997: 221-230.
16 McKee KK, Tan CP, Palyha OC, et al. Cloning and characterization of two human G protein-coupled receptor genes (GPR38 and GPR39) related to the growth hormone secretagogue and neurotensin receptors. Genomics, 1997, 46: 426-434.
17 Ong H, McNicoll N, Escher E, et al. Identification of a pituitary growth hormone-releasing peptide (GHRP) receptor subtype by photoaffinity labeling. Endocrinology, 1998, 139: 432-435.
18 Muccioli G, Ghe C, Ghigo MC, et al. Specific receptors for synthetic GH secretagogues in the human brain and pituitary gIand. J Endocrinol, 1998, 157: 99-106.
19 De Gennaro Colonna V, Rossoni G, Bernareggi M, et al. Cardiac ischemia and impairment of vascular endothelinm function in hearts from growth hormone-deficient rats: protection by hexarelin. Eur J Pharmacol, 1997, 334: 201-207.
20 Tan CE, McKee KK, Lin O, et al. Cloning and characterisation of a human and murine T-cell orphan G-protein-coupled receptor similar to the growth hormone secretagogue and neurotensin receptors. Genomics, 1998, 52: 223-229.
21 Cheng K, Chan W-W, Barreto A, et al. The synergistic effects of His-D-Trp-Ala- Trp-D-Phe-Lys-NH2 on growth hormone (GH)-releasing factor-stimulated GH release and intracellular adenosin 3', 5’-monophosphate accumulation in rat pituitary cell culture. Endocrinology, 1989, 124: 2791-2798.
22 Cheng K, Chan WW, Butler B, et al. Evidence for a role of protein kinase-C in His-D-Trp-Ala-Trp-D-Phe- Lys-NH2-induced growth hormone release from rat primary pituitary cells. Endocrinology, 1991, 129: 3337-3342.
23 Akman MS, Girard M, O'Brien LE Ho AK, et al. Mechanisms of action of a second generation growth hormone-releasing peptide (Ala-His-D-beta-Nal-Ala-Trp- D-Phe-Lys-NH2) in rat anterior pituitary cells. Endocrinology, 1993, 132: 1286-1291.
24 Pong SS, Chaung LY, Smith RG. GHRP-6 (His-D-Trp-Ala-Trp- D-Phe-Lys-NH2) stimulates growth hormone secretion by depolarization in rat pituitary cell cultures.
73rd Annual Meeting of the Endocrine Society, Washington, 1991.
25 Herrington J, Hille B. Growth hormone-releasing hexapeptide elevates intracellular calcium in rat somatotropes by 2 mechanisms. Endocrinology, 1994, 135: 1100-1108.
26 Wu D, Chen C, Katoh K, et al. The effect of GH-releasingpeptide- 2 (GHRP-2 or KP 102) on GH secretion from primary cultured ovine pituitary ceils can be abolished by a specific GH-releasing factor (GRF) receptor antagonist. J Endocrinol, 1994, 140: R8-R13.
27 Cheng K, Chart WW, Butler B, et al. Stimulation of growth hormone release from rat primary pituitary cells by L-692,429, a novel non-peptidyl GH secretagogue. Endocrinology, 1993, 132: 2729-2731.
28 Adams EE, Lei T, Bucherfelder M, et al. Protein kinase C-dependent growth hormone releasing peptides stimulate cyclic adenosine 3', 5’-monophosphate production by human pituitary somatotropinomas expressing gsp oncogenes: evidence for cross-talk between transduction pathways. Mol Endocrinol, 1996, 10: 432-438.
29 Bresson-Bepoldin L, Odessa MF, Dufy-Barbe L. GHRP-6 stimulates calcium increase and growth hormone release in human somatotrophs in vivo. Endocrine, 1994, 2: 793-803.
30 Adams EE, Lei T, Buchfelder M, et al. Biochemical characteristics of human pituitary somatotrophinomas with or without gsp mutations. J Clin Endocrinol Metab, 1995, 80: 2077-2081.
31 Adams EF, Buchfelder M, Lei T, et al. In vitro responses of GH-secreting tumours with and without gsp oncogenes to octreotide, GHRH and growth hormone-releasingpeptide. In: von Werder K, Fahlbusch Reds. Pituitary adenomas: from basic research to diagnosis and therapy. Amsterdam: Elsevier, 1996: 43-47.
32 Korborins M, Adams EF, Grossman AB. The effects of GHsecretagognes on human pituitary cells in culture and on rat hypothalamic tissue. In: Dieguez C, Ghigo E, Boghen M, Casanueva FF, eds. Growth hormone secretagogues: basic findings and clinical implications. Rotterdam: Elsevier, 1998 (in press).
33 Lei T, Bucherfelder M, Fahlbush R, et al. Growth hormonereleasing peptide (GHRP-6) stimulates phosphatidylinositol (PI) turnover in human pituitary somatotroph cells. J Mol Endocrinol, 1995, 14: 135-138.
34 Houslay MD. Crosstalk: a pivotal rote for the protein kinase C in modulating relationships between signal transduction pathways. J Biol Chem, 1998, 264: 8802-8810.
35 Cheng J, Wu TJ, Butler B, et al. Growth hormone releasing peptides: a comparison of the growth hormone releasing activities of GHRP-2 and GHRP-6 in rat primary pituitary cells. Life Sci, 1997, 60: 1385-1392.
36 Hickey GJ, Jacks TM, Schleim KD, et al. Repeat administration of the GH secretagogue MK-0677 increases and maintains elevated IGE-I levels in beagles. J Endocrinol, 1997, 152: 183-192.
37 Carmignac DF, Bennett PA, Robinson ICAE. Effects of growth hormone secretagogues on prolactin release in anesthetized dwarf (dw/dw) rats. Endocrinology, 1998, 139: 3590-3596.
38 Arvat E, Gianotti L, Broglio F, et al. Oestrogen replacement does not restore the reduced GH-releasing activity of hexarelin, a synthetic hexapeptide, in post-menopausal women. Eur J Endocrinol, 1997, 136: 483-487.
39 Korbonits M, Jacobs RA, Aylwin S, et al. Expression of the growth hormone secretagogue receptor in pituitary adenomas and other neuroendocrine tumors. J Clin Endoctinol Metab, 1998, 83: 3624-3630.
40 Haddad G, Penabad JL, Bashey HM, et al. Expression of activin/inhibin subunit messenger ribonucleic acids by gonadotroph adenomas. J Clin Endocrinol Metab, 1994, 79: 1399-1403.
41 Renner U, Brockmeier S, Strasburger CJ, et al. Growth-hormone (GH)-releasing peptide stimulation of GH release from human somatotroph adenoma cells - interaction with GH-releasing hormone, thyrotropin-releasing-hormone, and octreotide. J Clin Endocrinol Metab, 1994, 78: 1090-1096.
42 Dekeyzer Y, Lenne E, Bertagna X. Widespread transcription of the growth hormone-releasing peptide receptor gene in neuroendocfine human tumors. Eur J Endocrinol, 1997, 137: 715-718.
43 Lania A, Ballar6 E, Corbetta S, et al. Growth hormone-releasing peptide (GHRP-6) increases intracellular calcium concentrations in cultured cells from human pituitary adenomas of different types. Eur J Endocrinol, 1998, 139: 343-348.
44 Adams EF, Huang B, Buchfelder M, et al. Presence of growth hormone secretagogue receptor messenger ribonucleic acid in human pituitary tumors and rat GH3 cells. J Clin EndocrinoI Metab, 1998, 83: 638-642.
45 Nielsen S, Mellemkjaer S, Rasmussen L, et al. Gene transcription of receptors for growth hormone-releasing peptide and somatostatin in human pituitary adenomas. J Clin Endocrinol Metab, 1998, 83: 2997-3000.
46 Barlier A, Grino M, Zamora AJ, et al. Expression of growth hormone secretagogue-receptors (GHS-R) in human pituitary adenomas. European Congress of Endocrinology, Seville, 1998: 332-341.
47 Jansson J, Svensson J, Bengtsson B, et al. Acromegaly and Cushing's syndrome due to ectopic production of GHRH and ACTH by a thymic carcinoid tumour: in vitro responses to GHRH and GHRP-6. Clin Endocrinol, 1998, 48: 243-250.
48 Hosoda H, Kojima M, Matsuo H, et al. Purification and characterization of rat des-Gln14-Ghrelin, a second endogenous ligand for the growth hormone secretagogue receptor. J Biol Chem, 2000, 275(29): 21995-22000.
49 Date Y, Kojima M, Hosoda H, et al. Ghrelin, a novel growth hormone-releasing acylated peptide, is synthesized in a distinct endocrine cell type in the gastrointestinal tracts of rats and humans. Endocrinology, 2000, 141(11): 4255-4261.
50 Nakazato M, Murakami N, Date Y, et al. A role for ghrelin in the central regulation of feeding. Nature, 2001, 409(6817): 194-198.
51 Nagaya N, Kojima M, Uematsu M, et al. Hemodynamic and hormonal effects of human ghrelin in healthy volunteers. Am J Physiol Regul Integr Comp Physiol, 2001, 280(5): 1483-1487.
52 Date Y, Murakami N, Kojima M, et al. Central effects of a novel acylated peptide, ghrelin on growth hormone release in rats. Biochem Biophys Res Commun, 2000, 275(2): 477-480.
53 Wren AM, Small CJ, Ward HL, et al. The novel hypothalamic peptide ghrelin stimulates food intake and growth hormone secretion. Endocrinology, 2000, 141(11): 4325-4328.
54 Shintani M, Ogawa Y, Ebihara K, et al. Ghrelin, an endogenous growth hormone secretagogue, is a novel orexigenic peptide that antagonizes leptin action through the activation of hypothalamic neuropeptide Y/Y1 receptor pathway. Diabetes, 2001, 50(2): 227-232.
55 Kojima M, Hosodo H, Date Y, et al. Ghrelin is a growth hormone releasing acylated peptide from stomach. Nature, 1999, 402(6762): 656-660.
56 Banks WA, Tschop M, Robinson SM, et al. Extent and direction of ghrelin transport across the blood–brain barrier is determined by its unique primary structure. J Pharmacol Exp Ther, 2002, 302(2): 822-827.
57 Bednarek MA, Feighner SD, Pong SS, et al. Structure–function studies on the new growth hormone-releasing peptide, ghrelin: minimal sequence of ghrelin necessary for activation of growth hormone secretagogue receptor 1a. J Med Chem, 2000, 43: 4370- 4376.
58 Muccioli G, Papotti M, Locatelli V, et al. Binding of 125I-labeled ghrelin to membranes from human hypothalamus and pituitary gland. J Endocrin Invest, 2001, 24: 7-9.
59 Date Y, Kojima M, Hosoda H, et al. Ghrelin, a novel growth hormone-releasing acylated peptide, is synthesised in a distinct endocrine cell type in the gastrointestinal tract of rats and humans. Endocrinology, 2000, 141: 4255-4261.
60 Seoane LM, Tovar S, Baldelli R, et al. Ghrelin elicits a marked stimulatory effect on GH secretion in freely-moving rats. Eur J Endocrinol, 2000, 143: 7-9.
61 Arvat E, Di Vito L, Broglio M, et al. Preliminary evidence that ghrelin, the natural GH secretagogue (GHS)-receptor ligand, strongly stimulates GH secretion in humans. J Endocrin Invest, 2000, 23: 493-495.
62 Peino R, Baldelli R, Rodriguez-Garcia J, et al. Ghrelin-induced growth hormone secretion in humans. Eur J Endocrinol, 2000, 143: 11-14.
63 Takaya K, Ariyasu H, Kanamoto N, et al. Ghrelin strongly stimulates growth hormone release in humans. J Clin Endocrinol Metab, 2000, 85: 4908-4911.
64 Kamegai J, Tamura H, Ishii S, Sugihara H, et al. Thyroid hormones regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2001, 13: 275-278.
65 Tamura H, Kamegai J, Sugihara H, et al. Glucocorticoids regulate pituitary growth hormone secretagogue receptor gene expression. J Neuroendocrinol, 2000, 12: 481- 485.
66 Petersenn S, Rasch AC, Penshorn M, et al. Genomic structure and transcriptional regulation of the human growth hormone secretagogue receptor. Endocrinology, 2001, 142: 2649-2659.
67 Katayama M, Nogami H, Nishiyama J, et al. Developmentally and regionally regulated expression of growth hormone secretagogue receptor mRNA in rat brain and pituitary gland. Neuroendocrinology, 2000, 72: 333-340.
68 Barlier A, Zamora AJ, Grino M, et al. Expression of functional growth hormone secretagogue receptors in human pituitary adenomas: polymerase chain reaction, triple in situ hybridization and cell culture studies. J Neuroendocrinol, 1999, 11: 491-502.
69 Smith RG, Feighner S, Prendergast K, et al. A new orphan receptor involved in pulsatile growth hormone release. Trends Endocrinol Metab, 1999, 10(4): 128-135.
70 Cheng K, Chan WWS, Butler B, et al. Evidence for a role of protein kinase-C in His-D-Trp-Ala-D-Phe-Lys-amide induced growth hormone release from rat pituitary cells. Endocrinology, 1991, 129: 3337-3342
71 Luton J, Cerdas S, Billaud L, et al. Clinical features of adrenocortical carcinoma, prognostic factors, and the effect of mitotane therapy. N Engl J Med, 1990, 322: 1195-1201.
72 Mazzocchi G, Neri G, Rucinski M, et al. Ghrelin enhances the growth of cultured human adrenal zona glomerulosa cells by exerting MAPK-mediated proliferogenic and antiapoptotic effects. Peptides, 2004, 25: 1269-1277.
73 Muccioli G, Pons N, Ghe C, et al. Ghrelin and des-acyl ghrelin both inhibit isoproterenol-induced lipolysis in rat adipocytes via a non-type 1a growth hormone secretagogue receptor. Eur J Pharmacol, 2004, 498: 27-35.
74 Nanzer AM, Khalaf S, Mozid AM, et al. Ghrelin exerts a proliferative effect on a rat pituitary somatotroph cell line via the mitogen-activated protein kinase pathway. Eur J Endocrinol, 2004, 151: 233-240.
75 Nogueiras R, Tovar S, Mitchell SE, et al. Regulation of growth hormone secretagogue receptor gene expression in the arcuate nuclei of the rat by leptin and ghrelin. Diabetes, 2004, 53: 2552-2558.
76 Thompson NM, Gill DA, Davies R, et al. Ghrelin and des-octanoyl ghrelin promote adipogenesis directly in vivo by a mechanism independent of GHS-R1a. Endocrinology, 2003, 145: 234-242.
77 Tortorella C, Macchi C, Spinazzi R, et al. Ghrelin, an endogenous ligand for the growth hormonesecretagogue receptor, is expressed in the human adrenal cortex. Int J Mol Med,2003, 12: 213-217.