Swope S L, Schonbrunn A
Proc Natl Acad Sci U S A. 1984 Mar;81(6):1822-6. doi: 10.1073/pnas.81.6.1822.
The amphibian tetradecapeptide, bombesin (BBS) has been shown to stimulate insulin secretion both in vivo and by pancreatic islet cells in vitro. To determine whether BBS can act directly on pancreatic beta cells, we examined its effects on insulin secretion by HIT-T15 cells (HIT cells), a clonal islet cell line. Addition of 100 nM BBS to HIT cells stimulated insulin release 25-fold within 30 sec. The rapid stimulatory effect of BBS on insulin release was short-lived: the secretory rate returned to basal levels after 90 min of BBS treatment. The decrease in the rate of insulin release in the continued presence of BBS was due not to depletion of intracellular insulin stores but to specific desensitization to this peptide. Stimulation of insulin secretion by BBS was dose dependent with an ED50 value (0.51 +/- 0.15 nM) similar to the concentration of BBS-like immunoreactive material in rat plasma. Five BBS analogs, including porcine gastrin-releasing peptide, were as powerful as BBS in stimulating insulin release. The relative potencies of the analogs tested indicated that the COOH-terminal octapeptide sequence in BBS was sufficient for stimulation of release. In contrast, 14 peptides structurally unrelated to BBS did not alter insulin secretion. BBS action was synergistic with that of glucagon; insulin secretion in the presence of maximal concentrations of both peptides was greater than the additive effects of the two peptides added individually. Somatostatin inhibited BBS-stimulated release by 69 +/- 1% with an ID50 value of 3.2 +/- 0.3 nM. These results show that BBS stimulation of insulin secretion by a clonal pancreatic cell line closely parallels its effects in vivo and support the hypothesis that BBS stimulates insulin secretion by a direct effect on the pancreatic beta cell. The clonal HIT cell line provides a homogeneous cell preparation amenable for studies on the biochemical mechanisms of BBS action in the endocrine pancreas.
两栖类十四肽铃蟾肽(BBS)已被证明在体内和体外均可刺激胰岛细胞分泌胰岛素。为了确定BBS是否能直接作用于胰腺β细胞,我们检测了其对克隆胰岛细胞系HIT-T15细胞(HIT细胞)胰岛素分泌的影响。向HIT细胞中添加100 nM BBS可在30秒内刺激胰岛素释放增加25倍。BBS对胰岛素释放的快速刺激作用是短暂的:BBS处理90分钟后,分泌率恢复到基础水平。在持续存在BBS的情况下胰岛素释放速率的降低并非由于细胞内胰岛素储备的耗尽,而是对该肽的特异性脱敏。BBS对胰岛素分泌的刺激呈剂量依赖性,其半数有效剂量(ED50)值(0.51±0.15 nM)与大鼠血浆中铃蟾肽样免疫反应性物质的浓度相似。包括猪胃泌素释放肽在内的五种BBS类似物在刺激胰岛素释放方面与BBS一样有效。所测试类似物的相对效力表明,BBS的COOH末端八肽序列足以刺激释放。相比之下,14种与BBS结构无关的肽并未改变胰岛素分泌。BBS的作用与胰高血糖素的作用具有协同性;在两种肽的最大浓度存在下的胰岛素分泌大于单独添加两种肽的相加效应。生长抑素以69±1%的幅度抑制BBS刺激的释放,半数抑制剂量(ID50)值为3.2±0.3 nM。这些结果表明,BBS对克隆胰腺细胞系胰岛素分泌的刺激与其在体内的作用密切平行,并支持BBS通过直接作用于胰腺β细胞来刺激胰岛素分泌的假说。克隆的HIT细胞系提供了一种均质的细胞制剂,适合用于研究BBS在内分泌胰腺中作用的生化机制。