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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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Effect of glibenclamide on the [Ca2+]i transient elevation induced by extracellular ATP.A, C, time series of the mean F ratio during the application of different agents. B, D, the maximal [Ca2+]i with different agents. Glibenclamide (100 µM) and 5-hydroxydecanoate (5-HD, 100 µM) decreased the ATP- or Tg-induced peak [Ca2+]i; pinacidil (100 µM) did not change the maximal [Ca2+]i; diazoxide (Dia, 100 µM) increased the peak [Ca2+]i without significant differences. * P<0.05, compared with the control. BS, bath solution.
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pone-0089083-g003: Effect of glibenclamide on the [Ca2+]i transient elevation induced by extracellular ATP.A, C, time series of the mean F ratio during the application of different agents. B, D, the maximal [Ca2+]i with different agents. Glibenclamide (100 µM) and 5-hydroxydecanoate (5-HD, 100 µM) decreased the ATP- or Tg-induced peak [Ca2+]i; pinacidil (100 µM) did not change the maximal [Ca2+]i; diazoxide (Dia, 100 µM) increased the peak [Ca2+]i without significant differences. * P<0.05, compared with the control. BS, bath solution.

Mentions: Glibenclamide (100 µM) decreased the peak of the [Ca2+]i transient elevation induced by extracellular ATP (100 µM) (P<0.05, Figure 3 A, B). Pinacidil (100 µM) and other unselective potassium channel blockers (TEA and 4-AP) did not change the maximal [Ca2+]i (Figure 3 A, B). The mitochondrial KATP blocker 5-HD (100 µM) also decreased the maximal [Ca2+]i (P<0.05, Figure 3 A, B). When simultaneously pretreated with glibenclamide (100 µM) and 5-HD (100 µM), the maximal [Ca2+]i transient was decreased to the same level as that by either glibenclamide or 5-HD alone (P>0.05, Figure 3 A, B).


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 [Ca2+]i transient elevation induced by extracellular ATP.A, C, time series of the mean F ratio during the application of different agents. B, D, the maximal [Ca2+]i with different agents. Glibenclamide (100 µM) and 5-hydroxydecanoate (5-HD, 100 µM) decreased the ATP- or Tg-induced peak [Ca2+]i; pinacidil (100 µM) did not change the maximal [Ca2+]i; diazoxide (Dia, 100 µM) increased the peak [Ca2+]i without significant differences. * P<0.05, compared with the control. BS, bath solution.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0089083-g003: Effect of glibenclamide on the [Ca2+]i transient elevation induced by extracellular ATP.A, C, time series of the mean F ratio during the application of different agents. B, D, the maximal [Ca2+]i with different agents. Glibenclamide (100 µM) and 5-hydroxydecanoate (5-HD, 100 µM) decreased the ATP- or Tg-induced peak [Ca2+]i; pinacidil (100 µM) did not change the maximal [Ca2+]i; diazoxide (Dia, 100 µM) increased the peak [Ca2+]i without significant differences. * P<0.05, compared with the control. BS, bath solution.
Mentions: Glibenclamide (100 µM) decreased the peak of the [Ca2+]i transient elevation induced by extracellular ATP (100 µM) (P<0.05, Figure 3 A, B). Pinacidil (100 µM) and other unselective potassium channel blockers (TEA and 4-AP) did not change the maximal [Ca2+]i (Figure 3 A, B). The mitochondrial KATP blocker 5-HD (100 µM) also decreased the maximal [Ca2+]i (P<0.05, Figure 3 A, B). When simultaneously pretreated with glibenclamide (100 µM) and 5-HD (100 µM), the maximal [Ca2+]i transient was decreased to the same level as that by either glibenclamide or 5-HD alone (P>0.05, Figure 3 A, B).

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