目的:探讨降钙素基因相关肽(CGRP)对大鼠胃平滑肌的调控作用。方法:贴块法原代培养大鼠胃平滑肌细胞,特异性平滑肌肌动蛋白a-actin免疫细胞化学鉴定。实验分CGRPI组(10-7mol/LCGRP)、CGRPⅡ组(10~mol/LCGRP)、CGRPI]]组(10-9^mol/LCGRP)和干预组(10-6mol/LCGRP8-37)与空白对照组。图像分析系统测定大鼠胃平滑肌细胞长度;Ca3+荧光指示剂测定细胞内Ca2+浓度;CellTiter-Glo试剂盒检测细胞内ATP含量。结果:CGRP可舒张胃平滑肌细胞,降低胃平滑肌细胞游离Ca2+浓度及ATP含量。结论:CGRP通过受体介导直接参与胃平滑肌细胞松弛;CGRP可能通过降低胃平滑肌细胞游离Ca3+浓度、ATP含量而发挥其调控作用。
Objective: To investigate regulation of the calcitonin gene-related peptide (CGRP) on the gastric smooth muscle in rats. Methods: Primary rat gastric smooth muscle cells were cultured using tissue attachment block method. Specific smooth muscle a-actin antibody was stained by immunohistochemistry to identify the primary cultured cells. There were 5 groups: CGRP group I , Ⅱ ,Ⅲ (10-7 , 10-8 , 10-9 mol/L)group, CGRP 8-37 group(10-6 mol/L) and the control group. The length of smooth muscle cells was measured by micrometry before and after giving CGRP; by means of fluorescent indicator Fura-2/ AM, Ca2+ were tested to show CGRP how to affect gastric smooth muscle cells; CellTiter-Glo Kit was used to detect intracellular ATP content. Results: Gastric smooth muscle cells of rats were isolated successfully. CGRP reduced gastric smooth muscle in various concentrations of free Ca2+ concentration and ATP content. Conclusion: CGRP is directly involved in the relaxation of gastric smooth muscle cells by receptor mediator; CGRP plays a regulatory role in: gastric smooth muscle cells by reducing the concentration of free Ca2+ and ATP content.