The hypothalamic Arg-Phe-amide-related peptides, gonadotropin-inhibitory hormone and orthologous mammalian peptides of Arg-Phe-amide, may be important regulators of the hypothalamus-pituitary-gonadal reproductive axis...The hypothalamic Arg-Phe-amide-related peptides, gonadotropin-inhibitory hormone and orthologous mammalian peptides of Arg-Phe-amide, may be important regulators of the hypothalamus-pituitary-gonadal reproductive axis. These peptides may modulate the effects of kisspeptins because they are presently recognized as the most potent activators of the hypothalamus-pituitary-gonadal axis. However, their effects on gonadotropin-releasing hormone neurons have not been investigated. In the current study, the GT1-7 cell line-expressing gonadotropin-releasing hormone was used as a model to explore the effects of Arg-Phe- amide-related peptides on kisspeptin activation. Intracellular calcium concentration was quantified using the calcium-sensitive dye, fura-2 acetoxymethyl ester. Gonadotropin-releasing hormone released into the medium was detected via enzyme-linked immunosorbent assay. Results showed that 100 nmol/L kisspeptin-10 significantly increased gonadotropin-releasing hormone levels （at 120 minutes of exposure） and intracellular calcium concentrations. Co-treatment of kisspeptin with 1 μmol/L gonadotropin-inhibitory hormone or 1 μmol/L Arg-Phe-amide-related peptide-1 significantly attenuated levels of kisspeptin-induced gonadotropin-releasing hormone but did not affect kisspeptin-induced elevations of intracellular calcium concentration. Overall, the results suggest that gonadotropin-inhibitory hormone and Arg-Phe-amide-related peptide-1 may have inhibitory effects on kisspeptin-activated gonadotropin-releasing hormone neurons independent of the calcium signaling pathway.展开更多
自组装的纳米纤维肽作为一种新型材料，被广泛应用于细胞的三维培养以及组织缺损的修复。本研究在传统自组装多肽RADA16—1（RAD16）的基础上，为成骨细胞体外培养构建了一种支架材料，即把细胞黏附基序arginine—glycine—aspartic（RG...自组装的纳米纤维肽作为一种新型材料，被广泛应用于细胞的三维培养以及组织缺损的修复。本研究在传统自组装多肽RADA16—1（RAD16）的基础上，为成骨细胞体外培养构建了一种支架材料，即把细胞黏附基序arginine—glycine—aspartic（RGD）连接到RAD16上，再与纯RAD16按比例混合，构成本实验的研究材料Hybrid。用原子力显微镜（atomic force microscope，AFM）观察材料的微观结构。通过相差显微镜、MTT、组织化学染色等工具或方法观察成骨细胞（MC3T3-E1）在材料中的生长、增殖和分化情况。结果显示，Hybrid可以形成良好的纳米纤维结构。与RAD16相比，这种改进的纳米材料可以促进细胞的增殖，且碱性磷酸酶（alkaline phosphatase，ALP）的表达呈强阳性，有明显的钙结节形成。实验表明该材料在骨组织工程方面有潜在的应用价值。展开更多
基金supported by a grant from TUBITAK,project No.110S381
文摘The hypothalamic Arg-Phe-amide-related peptides, gonadotropin-inhibitory hormone and orthologous mammalian peptides of Arg-Phe-amide, may be important regulators of the hypothalamus-pituitary-gonadal reproductive axis. These peptides may modulate the effects of kisspeptins because they are presently recognized as the most potent activators of the hypothalamus-pituitary-gonadal axis. However, their effects on gonadotropin-releasing hormone neurons have not been investigated. In the current study, the GT1-7 cell line-expressing gonadotropin-releasing hormone was used as a model to explore the effects of Arg-Phe- amide-related peptides on kisspeptin activation. Intracellular calcium concentration was quantified using the calcium-sensitive dye, fura-2 acetoxymethyl ester. Gonadotropin-releasing hormone released into the medium was detected via enzyme-linked immunosorbent assay. Results showed that 100 nmol/L kisspeptin-10 significantly increased gonadotropin-releasing hormone levels （at 120 minutes of exposure） and intracellular calcium concentrations. Co-treatment of kisspeptin with 1 μmol/L gonadotropin-inhibitory hormone or 1 μmol/L Arg-Phe-amide-related peptide-1 significantly attenuated levels of kisspeptin-induced gonadotropin-releasing hormone but did not affect kisspeptin-induced elevations of intracellular calcium concentration. Overall, the results suggest that gonadotropin-inhibitory hormone and Arg-Phe-amide-related peptide-1 may have inhibitory effects on kisspeptin-activated gonadotropin-releasing hormone neurons independent of the calcium signaling pathway.
文摘自组装的纳米纤维肽作为一种新型材料，被广泛应用于细胞的三维培养以及组织缺损的修复。本研究在传统自组装多肽RADA16—1（RAD16）的基础上，为成骨细胞体外培养构建了一种支架材料，即把细胞黏附基序arginine—glycine—aspartic（RGD）连接到RAD16上，再与纯RAD16按比例混合，构成本实验的研究材料Hybrid。用原子力显微镜（atomic force microscope，AFM）观察材料的微观结构。通过相差显微镜、MTT、组织化学染色等工具或方法观察成骨细胞（MC3T3-E1）在材料中的生长、增殖和分化情况。结果显示，Hybrid可以形成良好的纳米纤维结构。与RAD16相比，这种改进的纳米材料可以促进细胞的增殖，且碱性磷酸酶（alkaline phosphatase，ALP）的表达呈强阳性，有明显的钙结节形成。实验表明该材料在骨组织工程方面有潜在的应用价值。
