title: Glibenclamide decreases ATP-induced intracellular calcium transient elevation via inhibiting reactive oxygen species and mitochondrial activity in macrophages.
abstract: Increasing evidence has revealed that glibenclamide has a wide range of anti-inflammatory effects. However, it is unclear whether glibenclamide can affect the resting and adenosine triphosphate (ATP)-induced intracellular calcium ([Ca(2+)]i) handling in Raw 264.7 macrophages. In the present study, [Ca(2+)]i transient, reactive oxygen species (ROS) and mitochondrial activity were measured by the high-speed TILLvisION digital imaging system using the indicators of Fura 2-am, DCFDA and rhodamine-123, respectively. We found that glibenclamide, pinacidil and other unselective K(+) channel blockers had no effect on the resting [Ca(2+)]i of Raw 264.7 cells. Extracellular ATP (100 µM) induced [Ca(2+)]i transient elevation independent of extracellular Ca(2+). The transient elevation was inhibited by an ROS scavenger (tiron) and mitochondria inhibitor (rotenone). Glibenclamide and 5-hydroxydecanoate (5-HD) also decreased ATP-induced [Ca(2+)]i transient elevation, but pinacidil and other unselective K(+) channel blockers had no effect. Glibenclamide also decreased the peak of [Ca(2+)]i transient induced by extracellular thapsigargin (Tg, 1 µM). Furthermore, glibenclamide decreased intracellular ROS and mitochondrial activity. When pretreated with tiron and rotenone, glibenclamide could not decrease ATP, and Tg induced maximal [Ca(2+)]i transient further. We conclude that glibenclamide may inhibit ATP-induced [Ca(2+)]i transient elevation by blocking mitochondria KATP channels, resulting in decreased ROS generation and mitochondrial activity in Raw 264.7 macrophages.
authors.affiliation: Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, Jinan, China ; Department of Neurology, Qilu Hospital, Shandong University, Jinan, China.
authors.forename: Duo-ling
authors.initials: DL
authors.lastname: Li
authors.affiliation: Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, Jinan, China.
authors.forename: Zhi-yong
authors.initials: ZY
authors.lastname: Ma
authors.affiliation: Department of Otorhinolaryngology, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan, China.
authors.forename: Zhi-jie
authors.initials: ZJ
authors.lastname: Fu
authors.affiliation: Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, Jinan, China.
authors.forename: Ming-ying
authors.initials: MY
authors.lastname: Ling
authors.affiliation: Department of Neurology, Qilu Hospital, Shandong University, Jinan, China.
authors.forename: Chuan-zhu
authors.initials: CZ
authors.lastname: Yan
authors.affiliation: Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education and Chinese Ministry of Public Health, Qilu Hospital, Shandong University, Jinan, China.
authors.forename: Yun
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D005452:Fluoresceins; D005453:Fluorescence; D016257:Fura-2 / Q000031:analogs & derivatives; D005905:Glyburide / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects*; D053078:Membrane Potential, Mitochondrial / Q000502:physiology; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology*; D020110:Pinacidil; D015221:Potassium Channels / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors*; D020112:Rhodamine 123; D012402:Rotenone
chemical_list: D005452:Fluoresceins; D015221:Potassium Channels; D017382:Reactive Oxygen Species; C117780:mitochondrial K(ATP) channel; D012402:Rotenone; C049925:fura-2-am; D020112:Rhodamine 123; C029569:diacetyldichlorofluorescein; D020110:Pinacidil; D000255:Adenosine Triphosphate; D005905:Glyburide; D002118:Calcium; D016257:Fura-2