eNampt, CRH and Il-6 influence on gene expression of rat primary corticotropes.

ACTH is known to be secreted from pituitary cortictropes after CRH and Il-6 stimulation. Recenetly it was proven that eNampt also stimulates ACTH secretion from rat pituitary gland. We used microarrays to detail the global programme of gene expression in isolated rat pituitary corticotropes, 24h after administration of eNampt, CRH and Il-6. Rat corticotropes were isolated from primary rat pituitary gland culture with anti-CRH antibody used as selection marker. Isolated Rat cortictropes were subjected to the cell culture. Subsequently eNampt, CRH and Il-6 were administered into the cell medium. After 24h, RNA were extracted and hybridize into the Affymetrix microarrays.

已知促肾上腺皮质激素（ACTH）可在促肾上腺皮质激素释放激素（CRH）与白细胞介素6（IL-6）的刺激下，由垂体促肾上腺皮质激素细胞分泌。近期已有研究证实，细胞外烟酰胺磷酸核糖转移酶（eNampt）同样可刺激大鼠垂体分泌ACTH。本研究采用微阵列（microarray）技术，对大鼠原代分离的垂体促肾上腺皮质激素细胞在给予eNampt、CRH及IL-6刺激24小时后的全基因表达谱进行了详细解析。实验中，我们以抗CRH抗体作为筛选标记，从大鼠原代垂体细胞培养物中分离得到促肾上腺皮质激素细胞。将分离得到的大鼠促肾上腺皮质激素细胞进行体外培养。随后向细胞培养基中加入eNampt、CRH与IL-6进行刺激。刺激24小时后，提取细胞总RNA，并将其与Affymetrix微阵列芯片进行杂交。