绵羊气管上皮细胞中环磷酸鸟苷的调节

Regulation of guanosine 3':5'-cyclic monophosphate in ovine tracheal epithelial cells.

作者信息

Range S P, Holland E D, Basten G P, Knox A J

机构信息

Division of Respiratory Medicine, City Hospital, Nottingham.

出版信息

Br J Pharmacol. 1997 Apr;120(7):1249-54. doi: 10.1038/sj.bjp.0701040.

DOI:10.1038/sj.bjp.0701040

PMID:9105699

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1564600/

Abstract

Guanosine 3':5'-cyclic monophosphate (cyclic GMP) is an important second messenger mediating the effects of nitric oxide (NO) and natriuretic peptides. Cyclic GMP pathways regulate several aspects of lung pathophysiology in a number of airway cells. The regulation of this system has not been extensively studied in pulmonary epithelial tissue. 2. We have studied the production of cyclic GMP by suspensions of ovine tracheal epithelial cells in response to activators of soluble guanylyl cyclase (sodium nitroprusside (SNP) and S-nitroso-N-acetyl-penicillamine (SNAP) and particulate guanylyl cyclase (atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), C-type natriuretic peptide (CNP) and E. coli heat stable enterotoxin (STa)). 3. Both 10(-7)-10(-3) M and 10(-7)-10(-3) M SNAP generated a concentration-dependent marked elevation in cyclic GMP production when incubated with 10(-3) M 3-isobutyl-l -methylxanthine (IBMX) (both greater than 25 x baseline values with highest drug concentration). 4. The increase in production of cyclic GMP in response to 10(-6) M SNP and 10(-5) M SNAP was markedly inhibited by both 5 x 10(-5) M haemoglobin (102% and 92% inhibition) and 5 x 10(-5) M methylene blue (82% and 84% inhibition). 5. The increase in cyclic GMP in response to 10(-3) M SNP was measured following co-incubation with the phosphodiesterase inhibitors 10(-7)-10(-3) M IBMX, 10(-7)-10(-4) M milrinone and 10(-7)-10(-4) M SKF 96231. Only 10(-4)-10(-3) M IBMX significantly increased cyclic GMP levels. 6. Cyclic GMP production was also significantly elevated from baseline by 10(-5) M ANP, 10(-5) M BNP, 10(-5) M CNP and 200 iu ml-3 of E. coli STa toxin in the presence of 10(-3) M IBMX. Increases with these natriuretic peptides and STa toxin were smaller in magnitude (2-4 fold) than those seen with SNP and SNAP. CNP was the most potent of the natriuretic peptides studied suggesting type B membrane bound guanylate cyclase is the predominant form expressed. 7. These results suggest that ovine tracheal epithelial cells contain active guanylyl cyclases. The more marked response to SNP and SNAP than to natriuretic peptides suggests that soluble guanylyl cyclase predominates.

摘要

鸟苷3':5'-环磷酸（环鸟苷酸，cGMP）是一种重要的第二信使，介导一氧化氮（NO）和利钠肽的作用。环鸟苷酸途径在多种气道细胞中调节肺病理生理学的多个方面。该系统在肺上皮组织中的调节尚未得到广泛研究。2. 我们研究了绵羊气管上皮细胞悬液对可溶性鸟苷酸环化酶激活剂（硝普钠（SNP）和S-亚硝基-N-乙酰青霉胺（SNAP））以及颗粒性鸟苷酸环化酶（心房利钠肽（ANP）、脑利钠肽（BNP）、C型利钠肽（CNP）和大肠杆菌热稳定肠毒素（STa））的反应中环鸟苷酸的产生。3. 当与10⁻³ M 3-异丁基-1-甲基黄嘌呤（IBMX）一起孵育时，10⁻⁷ - 10⁻³ M的SNP和10⁻⁷ - 10⁻³ M的SNAP均能引起环鸟苷酸产生的浓度依赖性显著升高（在最高药物浓度下均大于基线值的25倍）。4. 5×10⁻⁵ M血红蛋白（抑制率分别为102%和92%）和5×10⁻⁵ M亚甲蓝（抑制率分别为82%和84%）均显著抑制了10⁻⁶ M SNP和10⁻⁵ M SNAP引起的环鸟苷酸产生增加。5. 在与磷酸二酯酶抑制剂10⁻⁷ - 10⁻³ M IBMX、10⁻⁷ - 10⁻⁴ M米力农和10⁻⁷ - 10⁻⁴ M SKF 962³共同孵育后，测定了10⁻³ M SNP引起的环鸟苷酸增加。只有10⁻⁴ - 1⁻³ M IBMX显著提高了环鸟苷酸水平。6. 在10⁻³ M IBMX存在的情况下，10⁻⁵ M ANP、10⁻⁵ M BNP、10⁻⁵ M CNP和200 iu/ml³的大肠杆菌STa毒素也使环鸟苷酸产生从基线显著升高。这些利钠肽和STa毒素引起的增加幅度（2 - 4倍）小于SNP和SNAP引起的增加幅度。CNP是所研究的利钠肽中最有效的，表明B型膜结合鸟苷酸环化酶是主要表达形式。7. 这些结果表明绵羊气管上皮细胞含有活性鸟苷酸环化酶。对SNP和SNAP的反应比对利钠肽的反应更明显，表明可溶性鸟苷酸环化酶占主导地位。