title: Change of calcium and cAMP concentration by adrenoceptor agents in cultured porcine corneal endothelial cells.
abstract: It has been reported that beta-adrenergic receptors are localized in the corneal endothelial cells. In this study, the change of cellular signal transduction, such as intracellular calcium and cAMP, was determined with pure adrenergic agonists and commercial antiglaucoma adrenergic agents. The intracellular calcium of cultured porcine corneal endothelial cells was inhibited by 10 microM isoproterenol and norepinephrine, but enhanced by propranolol and 50 mM KCl. In the case of phenylephrine, calcium mobility did not alter significantly. Verapamil, at 10 microM, decreased intracellular calcium concentration. In the presence of isoproterenol, cellular cAMP concentration increased from 28.8 pmole/mg protein (1 microM) to 42.2 pmole/mg protein (100 microM) compared with control of 6.07 pmole/mg protein. Incubation with commercial adrenergic eye drops, such as betaxolol, caused the cAMP concentration to increase from 21.6 pmole/mg protein (0.0005%) to 39.1 pmole/mg protein (0.05%). Adding commercial levobunolol and timolol into cells caused cellular cAMP to increase from 14.3 pmole/mg protein (0.0005%) to 840.5 pmole/mg protein (0.05%) and from 115.2 pmole/mg protein (0.00025%) to 931.0 pmole/mg protein (0.025%), respectively. However, the preservative, benzalkonium chloride, increased cellular cAMP from 15.4 pmole/mg protein (0.00001 mg/ml) to 1087.4 pmole/mg protein (0.01 mg/ml). It is concluded that the intracellular calcium of corneal endothelium decreases when the cellular adrenergic receptor is activated by agonists. Benzalkonium chloride, due to its preservative in commercial antiglaucoma agents which increases cellular cAMP, may alter corneal endothelial physiology through long-term use.
authors.affiliation: Department of Ophthalmology, Kaohsiung Medical College, Taiwan, Republic of China.
authors.forename: H Z
authors.initials: HZ
authors.lastname: Wang
authors.forename: S J
authors.initials: SJ
authors.lastname: Hong
authors.forename: K Y
authors.initials: KY
authors.lastname: Wu
mesh_terms: D000322:Adrenergic Agonists / Q000494:pharmacology*; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology*; D000818:Animals; D001548:Benzalkonium Compounds / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism*; D004728:Endothelium, Corneal / Q000187:drug effects* / Q000378:metabolism; D011941:Receptors, Adrenergic / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D013552:Swine
chemical_list: D000322:Adrenergic Agonists; D000319:Adrenergic beta-Antagonists; D001548:Benzalkonium Compounds; D011941:Receptors, Adrenergic; D000242:Cyclic AMP; D002118:Calcium