【摘要】 目的分析胰淀素受体在胰淀素抑制胰岛B细胞功能中的作用。 方法 胰淀素、胰淀素受体拮抗剂AC253单独及联合短时间作用于大鼠胰岛B细胞系INS-1细胞后，ELISA及RT-PCR检测葡萄糖刺激胰岛素分泌及mRNA转录情况，共聚焦显微镜下观察高糖、磺脲类药物及KCl刺激下细胞内Ca2+浓度的变化，RT-PCR检测胰淀素短时间作用对胰淀素受体成分受体活性修饰蛋白(RAMP)rnJRNA表达的影响。 结果胰淀素短时间孵育高糖刺激下胰岛素分泌由(2.08±0.35)ng／ml降至(0.73±0．24)ng／ml (P<0.01)，mRNA转录由(1.00±0.12)降至(0.39±0．09)(P<0.01)，加用AC253后胰岛素分泌升至(1-41±0.32)ng／ml，mRNA转录升至(0.62±0.08)，与单用胰淀素组比较，差异有统计学意义(P<0.05)。AC253可缓解胰淀素对高糖刺激下Ca抖升高的抑制作用，Ca2+总体变化水平和峰值由(650．155±4．818)和(1．154±0．011)升至(769．795±4．956)和(1．589±0.013) (P均<0.01)，同样AC253也可缓解胰淀素对磺脲类药物及KCl刺激下细胞内Ca^2＋升高的抑制作用。胰淀素短时间孵育未改变RAMP mRNA的表达。 结论胰淀素受体可能参与了胰淀素抑制胰岛G细胞的胰岛素分泌、合成及对细胞内Ca2+浓度的调节。
    【关键词】胰淀素；胰淀素受体；胰岛B细胞；胰岛素分泌

    【Abstract】  Objective   To investigate the effects of amylin receptor on inhibition of pancreatic p cell function induced by amylin． Methods   Amylin and specific amylin receptor antagonist AC253 were applied to INS-1 cells separately or simultaneously．Glucose-stimulated insulin release and mRNAtranscription were measured by ELISA and RT-PCR respectively．Laser scanning confocal microscope wasused to monitor the changes of intracellular fluorescence intensity which represent Ca²⁺ concentration changes upon glucose、sulfonylurea or KCl stimulation．The influence of short term incubation with amylin on mRNA expression of receptor activity modifying protein(RAM[P)as a component of amylin reccptor was observed by RT—PCR． Results Glucose-stimulated insulin secretion was decreased from(2.08±0.35)ng／ml to(0.73±0.24) ng/ml  (P<0.01)，and rnRNA transcription was decreased from(1.00±0.12)to(0.39±0.09)(P<0.01)by short time exposure to amylin．Insulin release was promoted to (1.41±0.32) ng/ml，and mRNA was increased to

(0.62±0.08) by addition of AC253，statistically different fromusing amylin alone(P<0.05)．AC253 attenuated amylin’S inhibition on glucose—induced increase of Ca²⁺，with average Of  Ca²⁺，and peak value increased from (650.155±4.818)and (1.154±0.011) to(769.795±4.956) and (1.589±0．013) (P<0.01)．Similarly。the inhibitory effects of amylin on calcium influx induced by sulfonylurea or KCl were alI attenuated in the presence of AC253．No changes were detected in RAMP mRNA expression after short term incubation with amylin． Conclusion   Amylin receptor may participate in amylin’S inhibition on insulin release，synthesis and regulation of intracellular Ca²⁺．
    【Key words】  Amylin；Amylin recept。r；Pancreatic B cell；Insulin secretion