ns19504  (Alomone Labs)

Journal: Scientific Reports

Article Title: Pharmacological activation of BK channels protects against LPS-induced pneumonia

doi: 10.1038/s41598-025-08902-6

Figure Lengend Snippet: Pharmacological BK channel activation protects mice against LPS-induced pneumonia. LPS infection ( i.t. , 10 mg/kg) increased BALF total cell ( a , k ) and neutrophil counts ( b , l ), and BALF CCL-2 ( c , n ), MIP-1α ( o ) and CXCL-10 ( p ) concentrations, lung injury scores ( d , e ), and cytosolic ROS production by BALF cells ( f , m ). Two doses of the BK activator NS1619 or NS19504 ( i.t. , 0.66 mg/kg), given at 0 and 24 h improved all measured markers of acute lung injury. In contrast, mitochondrial ROS production ( g ), BALF total protein levels ( h , q ), quasi-static lung compliance ( i , r ), and body weight loss ( j , s ) were not affected by NS1619 treatment. Control mice received equimolar drug vehicle injections. n = 3–10 mice per group; individual experimental data points are depicted within each bar; bars depict mean ± SEM; *p < 0.05; IF—intensity of fluorescence; A.U.—arbitrary units; scale bar: 650 µm.

Article Snippet: Separate groups also received i.t. injections of the BK channel activator NS1619 (0.66 mg/kg in 50 μl in sterile PBS; Millipore, Burlington, MA) or NS19504 (1.33 mg/kg in 50 μl in sterile PBS; Alomone labs, Israel), the BK channel blocker Paxilline (1.33 mg/kg in 50 μl in sterile PBS; Alomone labs, Israel), or equimolar vehicle controls, at times 0 and 24 h. We then quantified the degree of lung injury at 48 h. This model resulted in a moderate degree of lung injury with potential for recovery (< 3% mortality), representing a clinically relevant infection.

Techniques: Activation Assay, Infection, Control, Fluorescence

Journal: Scientific Reports

Article Title: Pharmacological activation of BK channels protects against LPS-induced pneumonia

doi: 10.1038/s41598-025-08902-6

Figure Lengend Snippet: Pharmacological activation of BK channels with NS1619 (30 μM) or NS19504 (30 μM) administered at 0 h protects primary human alveolar epithelial cells (HPAEpiC against LPS (2 μg/ml) -induced ROS production ( a ). In contrast, primary human pulmonary artery endothelial cells (HPAEC) showed no ROS response to LPS treatment ( b ). Control cells were treated with an equimolar drug vehicle. n = 3–7 separate experimental repeats; individual experimental data points are shown within each bar; bars represent mean ± SEM; * p < 0.05; IF – intensity of fluorescence; A.U.—arbitrary units.

Article Snippet: Separate groups also received i.t. injections of the BK channel activator NS1619 (0.66 mg/kg in 50 μl in sterile PBS; Millipore, Burlington, MA) or NS19504 (1.33 mg/kg in 50 μl in sterile PBS; Alomone labs, Israel), the BK channel blocker Paxilline (1.33 mg/kg in 50 μl in sterile PBS; Alomone labs, Israel), or equimolar vehicle controls, at times 0 and 24 h. We then quantified the degree of lung injury at 48 h. This model resulted in a moderate degree of lung injury with potential for recovery (< 3% mortality), representing a clinically relevant infection.

Techniques: Activation Assay, Control, Fluorescence

Journal: Scientific Reports

Article Title: Pharmacological activation of BK channels protects against LPS-induced pneumonia

doi: 10.1038/s41598-025-08902-6

Figure Lengend Snippet: ( a ) BK channel-mediated network analysis predicts cytokine and ROS clusters connected through hydrogen peroxide (H 2 O 2 ) in an in silico model. A protein-drug interaction network was constructed using the STITCH database to investigate the relationships between BK channels, oxidative stress, and inflammatory molecules in an LPS-induced pneumonia model. The network includes BK channels (KCNMA1), BK channel activators (NS1619/NS19504), neutrophil/macrophage chemoattractants (CCL-2, MIP-1α, CXCL-10), and oxidative stress enzymes (SOD1, catalase). The network analysis identified two distinct clusters: a ROS cluster (including SOD2, zinc, and copper) and a cytokine cluster (including CCL-4, CXCR-2, CCR-2, CCL-5, and CXCL-9). These clusters are interconnected by H 2 O 2 , linking ROS production and cytokine release to BK channels in LPS-induced pneumonia. Dashed lines represent connections identified in this study; red nodes indicate query proteins obtained in this study, while gray nodes represent the most likely downstream interaction partners with a high confidence level (0.95 on a 0–1 scale). ( b ) Schematic diagram of the proposed mechanism underlying BK channel activation-mediated protection against LPS-induced oxidative stress.

Article Snippet: Separate groups also received i.t. injections of the BK channel activator NS1619 (0.66 mg/kg in 50 μl in sterile PBS; Millipore, Burlington, MA) or NS19504 (1.33 mg/kg in 50 μl in sterile PBS; Alomone labs, Israel), the BK channel blocker Paxilline (1.33 mg/kg in 50 μl in sterile PBS; Alomone labs, Israel), or equimolar vehicle controls, at times 0 and 24 h. We then quantified the degree of lung injury at 48 h. This model resulted in a moderate degree of lung injury with potential for recovery (< 3% mortality), representing a clinically relevant infection.

Techniques: In Silico, Construct, Activation Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: BK Channel-Mediated Microglial Phagocytosis Alleviates Neurological Deficit After Ischemic Stroke

doi: 10.3389/fncel.2021.683769

Figure Lengend Snippet: BK channel activator NS19504 promoted microglial phagocytosis after OGD in primary culture . (A) Representative images of beads (red) phagocytosed by CD11b + microglia (green) after 1 h of OGD. Control group: primary microglia cultured with basic microglia medium; NS19504 group: primary microglia cultured with 10 μM NS19504 in basic microglia medium; Paxilline group: primary microglia cultured with 1 μM Paxilline in basic microglia medium. Scale bar = 25 μm. (B) Bar graph showed that semiquantitative data of the relative bead density versus microglia. Data are mean + SE, n = 3 per group. * p < 0.05, control vs. NS19504 or Paxilline vs. NS19504 group. (C) Representative photomicrographs showed that neuronal debris (red) phagocytosed by microglia (green) in the NS19504- and Paxilline-treated mice following tMCAO. Scale bar = 25 μm. (D) Bar graph shows semiquantitative data of the proportion of microglia that phagocytose neuronal debris. Data are mean + SE, n = 3 per group. * p < 0.05, NS19504 vs. control or Paxilline vs. NS19504 group. (E) Representative photomicrographs show FITC and PE-positive cells. FITC-labeled beads were phagocytosed by PE-labeled microglia. Primary microglia were divided into the control group (basic microglia medium); LPS group (200 ng/ml LPS); LPS + NS19504 group (200 ng/ml LPS and 10 μM NS19504); and LPS + Paxilline group (200 ng/ml LPS and 1 μM Paxilline). (F) Bar graph shows semiquantitative analysis of the proportion of microglia that phagocytose beads. Data are mean + SE, n = 3 per group. ** p < 0.01, compared with the control group. # p < 0.05, compared with the Paxilline group. (G) Bar graph shows the result of CCK-8 assay in the primary microglia without or with different concentrations of Paxilline and NS19504 for 12 h of OGD. Data are mean ± SE, n = 3 per group. # p < 0.05, ## p < 0.01, compared with the control group. * p < 0.05, ** p < 0.01; NS represents for NS19504, and Pax represents for Paxilline.

Article Snippet: The selective BK channel antagonist Paxilline ( ) and activator NS19504 ( ) powder (Alomone Labs, Jerusalem, Israel) were, respectively, dissolved in DMSO to the final storage concentrations of 2 and 50 mM.

Techniques: Control, Cell Culture, Labeling, CCK-8 Assay

Journal: Frontiers in Cellular Neuroscience

Article Title: BK Channel-Mediated Microglial Phagocytosis Alleviates Neurological Deficit After Ischemic Stroke

doi: 10.3389/fncel.2021.683769

Figure Lengend Snippet: Activation of BK channels promoted the phagocytic function of microglia after tMCAO and decreased neuronal apoptosis . (A) Representative photomicrograms of IBA1 + microglia (green) and TUNEL + apoptotic cells (red) in the peri-infarct region of brain slice in the DMSO, NS19504, and Paxilline groups at 3 days after tMCAO. Scale bar = 50 μm. (B) Bar graph shows quantitative analysis of the number of IBA1 + TUNEL + cells per field. Data are mean + SE, n = 3 per group. *vs. DMSO group, ** p < 0.01. (C) Representative photomicrograms of NeuN + neurons (green) and TUNEL + apoptotic cells (red) in the peri-infarct region of brain slice in the DMSO, NS19504, and Paxilline groups at 3 days after tMCAO. Scale bar = 50 μm. (D) Bar graph shows quantitative analysis of the number of NeuN + /TUNEL + cells per vision. Data are mean + SE, n = 3 per group. *vs. DMSO group, ** p < 0.01.

Article Snippet: The selective BK channel antagonist Paxilline ( ) and activator NS19504 ( ) powder (Alomone Labs, Jerusalem, Israel) were, respectively, dissolved in DMSO to the final storage concentrations of 2 and 50 mM.

Techniques: Activation Assay, TUNEL Assay, Slice Preparation

Journal: Frontiers in Cellular Neuroscience

Article Title: BK Channel-Mediated Microglial Phagocytosis Alleviates Neurological Deficit After Ischemic Stroke

doi: 10.3389/fncel.2021.683769

Figure Lengend Snippet: Activation of BK channels ameliorated stroke outcomes. (A) Experiment design of the animal experiment. Three days before tMCAO, mice were subjected behavior training. After a 90-min tMCAO surgery, mice were divided randomly into four groups (sham, DMSO, NS19504, and Paxilline). At 1, 3, 7, and 14 days, behavior tests were performed, and protein and RNA samples were collected. (B) Weight changes of mice in three groups at 1, 2, 3, 7, and 14 days after tMCAO. (C) Modified Neurological Severity Score of three groups at 1, 3, 7, and 14 days after tMCAO. (D) Average score per second of three groups of hinging wire at 3 days after tMCAO. (E) Survivorship curve during 14 days of three groups after tMCAO. Data are mean + SE, n = 21–23 per group at 1 day, n = 17–23 per group at 3 days, n = 8–13 per group at 7 days, n = 3–9 per group at 14 days. *NS19504 vs. Paxilline group, ** p < 0.01. # NS19504 vs. DMSO, # p < 0.05.

Article Snippet: The selective BK channel antagonist Paxilline ( ) and activator NS19504 ( ) powder (Alomone Labs, Jerusalem, Israel) were, respectively, dissolved in DMSO to the final storage concentrations of 2 and 50 mM.

Techniques: Activation Assay, Modification

Journal: Frontiers in Cellular Neuroscience

Article Title: BK Channel-Mediated Microglial Phagocytosis Alleviates Neurological Deficit After Ischemic Stroke

doi: 10.3389/fncel.2021.683769

Figure Lengend Snippet: Inflammatory factors and phosphorylated-ERK expressions under different treatments at 3 days after tMCAO . (A) Bar graph represents the relative mRNA levels of inflammatory factors including TNF-α, TGF-β, IL-10, IL-6, IL-1β, and IL-1α. Data are mean + SE, n = 3 per group. (B) Representative image of Western blot of phosphorylated-ERK1/2 (p-ERK), total-ERK1/2 (t-ERK), and β-actin at 3 days after tMCAO. (C) Bar graph showing the relative protein levels of p -ERK vs. t-ERK at 3 day after tMCAO. Data are mean + SE, n = 9 per group. *NS19504 vs. Paxilline group, * p < 0.05; ns, no significance.

Article Snippet: The selective BK channel antagonist Paxilline ( ) and activator NS19504 ( ) powder (Alomone Labs, Jerusalem, Israel) were, respectively, dissolved in DMSO to the final storage concentrations of 2 and 50 mM.

Techniques: Western Blot