| B radykinin Receptor-Meadiated Intracellular Calcium Mobilization and ATP Changes in Cultured Bovine Corneal Endothelial Cell. |
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Suk Woo Yang, Seok Ho Cha, Keon Haeng Lee, Tae Won Hahn |
1Department of Ophthalmology, Kangnam St.Mary Hospital, College of Medicine, The Catholic University of Korea.
2Department of Pharmacology, College of Medicine, The Catholic University of Korea, Seoul, Korea. |
| 배양된 소 각막내피세포에서 Bradykinin 수용기 매개에 의한 세포내 칼슘 동원과 ATP 변화 |
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한태원(Tae Won Hahn),양석우(Suk Woo Yang),차석호(Seok Ho Cha),이권행(Keon Haeng Lee) |
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| Abstract |
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To clarify the effect of bradykinin(Bk)on cultured bovine corneal endothelial cells(BCEC), cytosolic free calcium([Ca2+])mobilization and cell proliferation were investigated. The [Ca2+] was determined using a Ca2+ sensitive indicator, Fura-2/AM, and cell proliferation was evaluated by counting the cell number. Bk induced the transient increase of [Ca2+] in a concentration-dependent manner(10(-11)M~10(-7)M)and its 50% effective concentration was about 5x10(-11)M. The basal [Ca2+] with 1mM CaCl2 in the bathing solution was 87+/-9nM. Transient Bk(10(-8)M)-induced [Ca2+] increase was inhibited slightly but significantly by the pretreatment with EGTA. The pretreatment with U-73122(5x10(-6)M), an inhibitor of phospholipase C, also attenuated Bkinduced [Ca2+] mobilization. To identify and characterize the Bk receptor subtype in BCEC, Bk1 and Bk2 antagonists were investigated. Transient Bk(10(-8)M)-induced [Ca2+] increase was almost absolutely attenuated by the pretreatment with Bk2 antagonist for 10 minutes. To investigate the physiological effect of Bk, Bk-induced mitogenic effect was studied. 10(-8)M of Bk produced significant increase of intracellular ATP levels from the day 2 to the day six of culture period. This Bk-induced mitogenic effect was inhibited by the treatment with Bk2 antagonist. Bk-induced ion transport was determined by measuring intracellular ATP contents. Intracellular ATP content([ATP]i)was decreased by the treatment with 10(-8)M Bk for 10 minutes. Bk-induced [ATP]i decrement was significantly restored by the pretreatment with ouabain for 30 minutes. In summary, stimulation of intracellular signal transduction by Bk in BCEC is coupled with Bk2 type receptor. And also, Bk produces mitogenic effect and enhancesion and fluid transport in BCEC. |
| Key Words:
Bovine corneal endothelial cells;Bradykinin;Bk2 receptor;Intracellular calcium;Intracellular ATP contents;Mitogenic effect |
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