Singh P, Asada I, Owlia A, Collins T J, Thompson J C
Department of Surgery, University of Texas Medical Branch, Galveston 77550.
Am J Physiol. 1988 Feb;254(2 Pt 1):G217-23. doi: 10.1152/ajpgi.1988.254.2.G217.
We have examined the direct effect of somatostatin (SRIF) on basal and stimulated amylase release from guinea pig pancreatic acini using the in vitro method of continuous perifusion. The optimal conditions of flow rate, chamber size, acinar cell volume per chamber, and period of secretagogue infusion were defined for the perifusion system. The kinetic profile of amylase release in response to cholecystokinin-octapeptide (CCK-8), vasoactive intestinal peptide (VIP), and SRIF was studied. Under optimal conditions, the acini were found to remain equally responsive to an ED50 dose of CCK-8 (0.5-0.8 nM) for 12 h of perifusion. The duration of amylase response to any given dose of CCK-8, given for the optimal period of 5 min, was 80-100 min. The total amylase released minus the basal release divided by 90 min (delta response) in response to the maximum effective (Maxeff) dose of CCK-8 (100 nM) was 14,667 +/- 1,433 U/l (amounting to a 10-fold increase compared with basal values). When compared with the amount of total delta amylase released in response to the Maxeff dose of CCK, the total amylase released in response to the Maxeff doses of SRIF (1 microM) and VIP (10 nM) was 10-21% and 51-59%, respectively. SRIF (100 nM) significantly decreased VIP- (0.1-1.0 nM) stimulated amylase release by 45-70% in the perifusion method of study but had no significant effect on the CCK-stimulated amylase release. This suggests that the perifusion method can be used for investigating the mechanism of SRIF-mediated inhibition of VIP effects on amylase release in an in vitro system.
我们采用连续灌流的体外方法,研究了生长抑素(SRIF)对豚鼠胰腺腺泡基础状态及刺激状态下淀粉酶释放的直接影响。确定了灌流系统的最佳流速、腔室大小、每个腔室的腺泡细胞体积以及促分泌素输注时间。研究了淀粉酶对八肽胆囊收缩素(CCK-8)、血管活性肠肽(VIP)和SRIF释放的动力学特征。在最佳条件下,发现腺泡在灌流12小时内对ED50剂量的CCK-8(0.5 - 0.8 nM)保持同等反应性。在最佳的5分钟给药期内,给予任何给定剂量CCK-8后淀粉酶反应的持续时间为80 - 100分钟。对CCK-8最大有效(Maxeff)剂量(100 nM)反应的总淀粉酶释放量减去基础释放量后除以90分钟(δ反应)为14,667 ± 1,433 U/l(与基础值相比增加了10倍)。与对CCK最大有效剂量反应释放的总δ淀粉酶量相比,对SRIF(1 μM)和VIP(10 nM)最大有效剂量反应释放的总淀粉酶量分别为10% - 21%和51% - 59%。在灌流研究方法中,SRIF(100 nM)显著降低VIP(0.1 - 1.0 nM)刺激的淀粉酶释放45% - 70%,但对CCK刺激的淀粉酶释放无显著影响。这表明灌流方法可用于在体外系统中研究SRIF介导的对VIP影响淀粉酶释放的抑制机制。