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Glibenclamide decreases ATP-induced intracellular calcium transient elevation via inhibiting reactive oxygen species and mitochondrial activity in macrophages.

Li DL, Ma ZY, Fu ZJ, Ling MY, Yan CZ, Zhang Y - PLoS ONE (2014)

Bottom Line: 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.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.Furthermore, glibenclamide decreased intracellular ROS and mitochondrial activity.

View Article: PubMed Central - PubMed

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.

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.

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Related in: MedlinePlus

Effect of glibenclamide on the resting [Ca2+]i of Raw 264.7 macrophages.A, representative calcium imaging of Raw 264.7 cells in control solution. B, representative calcium imaging with glibenclamide treatment (100 µM). C, glibenclamide has no effect on the resting [Ca2+]i with or without calcium in the extracellular solution. D, pinacidil and other unselective potassium channel blockers (4-aminopyridine, 4-AP, 10 µM; tetraethylammonium, TEA, 100 µM) did not change the resting [Ca2+]i.
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pone-0089083-g001: Effect of glibenclamide on the resting [Ca2+]i of Raw 264.7 macrophages.A, representative calcium imaging of Raw 264.7 cells in control solution. B, representative calcium imaging with glibenclamide treatment (100 µM). C, glibenclamide has no effect on the resting [Ca2+]i with or without calcium in the extracellular solution. D, pinacidil and other unselective potassium channel blockers (4-aminopyridine, 4-AP, 10 µM; tetraethylammonium, TEA, 100 µM) did not change the resting [Ca2+]i.

Mentions: Glibenclamide (100 µM) had no effect on the resting [Ca2+]i of Raw 264.7 cells with or without Ca2+ in extracellular solution (P>0.05, Figure 1A, C). Membrane KATP opener (pinacidil, 100 µM) and other unselective potassium channel blockers (TEA, 100 µM; 4-AP,10 µM) did not change the resting [Ca2+]i (P>0.05, Figure 1B, D).


Glibenclamide decreases ATP-induced intracellular calcium transient elevation via inhibiting reactive oxygen species and mitochondrial activity in macrophages.

Li DL, Ma ZY, Fu ZJ, Ling MY, Yan CZ, Zhang Y - PLoS ONE (2014)

Effect of glibenclamide on the resting [Ca2+]i of Raw 264.7 macrophages.A, representative calcium imaging of Raw 264.7 cells in control solution. B, representative calcium imaging with glibenclamide treatment (100 µM). C, glibenclamide has no effect on the resting [Ca2+]i with or without calcium in the extracellular solution. D, pinacidil and other unselective potassium channel blockers (4-aminopyridine, 4-AP, 10 µM; tetraethylammonium, TEA, 100 µM) did not change the resting [Ca2+]i.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928368&req=5

pone-0089083-g001: Effect of glibenclamide on the resting [Ca2+]i of Raw 264.7 macrophages.A, representative calcium imaging of Raw 264.7 cells in control solution. B, representative calcium imaging with glibenclamide treatment (100 µM). C, glibenclamide has no effect on the resting [Ca2+]i with or without calcium in the extracellular solution. D, pinacidil and other unselective potassium channel blockers (4-aminopyridine, 4-AP, 10 µM; tetraethylammonium, TEA, 100 µM) did not change the resting [Ca2+]i.
Mentions: Glibenclamide (100 µM) had no effect on the resting [Ca2+]i of Raw 264.7 cells with or without Ca2+ in extracellular solution (P>0.05, Figure 1A, C). Membrane KATP opener (pinacidil, 100 µM) and other unselective potassium channel blockers (TEA, 100 µM; 4-AP,10 µM) did not change the resting [Ca2+]i (P>0.05, Figure 1B, D).

Bottom Line: 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.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.Furthermore, glibenclamide decreased intracellular ROS and mitochondrial activity.

View Article: PubMed Central - PubMed

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.

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.

Show MeSH
Related in: MedlinePlus