Persaud S J, Jones P M
Biomedical Sciences Division, King's College London, Campden Hill Road, W8 7AH, London, UK.
Endocrine. 1995 Apr;3(4):285-9. doi: 10.1007/BF03021407.
The involvement of the family of protein kinase C (PKC) isoenzymes in the secretory response of rat islets of Langerhans to glucose, the major insulin secretagogue, was investigated using the PKC inhibitor Ro 31-8220, a derivative of staurosporine. Ro 31-8220 was a more selective PKC inhibitor than staurosporine in islets, having minimal effects on protein kinases activated by cyclic AMP or by Ca(2+) and calmodulin. The secretory response to 4βPMA, an activator of phorbol ester-sensitive isoforms of PKC, was abolished by Ro 31-8220. Basal insulin secretion (2MM: glucose) was not affected by Ro 31-8220, but 20MM: glucose-induced insulin release was inhibited in a dose-dependent manner, maximally by ∼50% at 10 µM: Ro 31-8220. Higher concentrations of Ro 31-8220 (507gmM: ) did not further inhibit the secretory response to glucose and also caused ∼50% inhibition of insulin secretion stimulated by 10MM: glyceraldehyde. Ca(2+)-stimulated insulin secretion from electrically permeabilised islets was not inhibited by Ro 31-8220. Calphostin C, which inhibits some isoforms of PKC by interacting with the diacylglycerol binding site, unexpectedly caused a large (∼10-fold) increase in secretion at 2MM: glucose, so could not be used in islets to further investigate the involvement of phorbol ester-sensitive PKC isoforms in the insulin secretory process. One possible explanation for our results using Ro 31-8220 is that phorbol ester-insensitive isoforms of PKC (ζ and/orι) are involved in glucose-stimulated insulin secretion from rat islets.
利用蛋白激酶C(PKC)抑制剂Ro 31-8220(一种星形孢菌素衍生物),研究了PKC同工酶家族在大鼠胰岛对主要胰岛素促分泌剂葡萄糖的分泌反应中的作用。在胰岛中,Ro 31-8220是比星形孢菌素更具选择性的PKC抑制剂,对由环磷酸腺苷或钙(Ca2+)及钙调蛋白激活的蛋白激酶影响极小。Ro 31-8220消除了对4βPMA(一种佛波酯敏感型PKC同工型的激活剂)的分泌反应。基础胰岛素分泌(2毫摩尔/升葡萄糖)不受Ro 31-8220影响,但20毫摩尔/升葡萄糖诱导的胰岛素释放受到剂量依赖性抑制,在10微摩尔/升Ro 31-8220时最大抑制约50%。更高浓度的Ro 31-8220(50微摩尔/升)并未进一步抑制对葡萄糖的分泌反应,还导致10毫摩尔/升甘油醛刺激的胰岛素分泌受到约50%的抑制。Ro 31-8220不抑制电透化胰岛中钙(Ca2+)刺激的胰岛素分泌。钙泊三醇C通过与二酰基甘油结合位点相互作用抑制某些PKC同工型,意外地在2毫摩尔/升葡萄糖时导致分泌大幅增加(约10倍),因此不能用于胰岛进一步研究佛波酯敏感型PKC同工型在胰岛素分泌过程中的作用。我们使用Ro 31-8220得到的结果的一种可能解释是,PKC的佛波酯不敏感型同工型(ζ和/或ι)参与了大鼠胰岛中葡萄糖刺激的胰岛素分泌。