Journal: International Journal of Molecular Sciences

Article Title: Role of Intracellular Na + in the Regulation of [Ca 2+ ] i in the Rat Suprachiasmatic Nucleus Neurons

doi: 10.3390/ijms20194868

Figure Lengend Snippet: Preferential enhancement by ouabain of nimodipine-insensitive Ca 2+ transients. ( A ) a representative experiment showing the effect of 2 µM nimodipine on 20 K + -induced Ca 2+ response (an average of 13 cells) in the absence and then presence of 10 µM ouabain. ( B,C ) Superimposition of Ca 2+ transients showing that ouabain inhibits nimodipine-sensitive ( B ) but enhances the nimodipine-insensitive ( C ) Ca 2+ transients. ( D ) Normalized Ca 2+ transients to show the delayed onset of Ca 2+ clearance (marked by horizontal bar) for ouabain-enhanced Ca 2+ transient in the absence (c – a) but not in the presence (d – b) of nimodipine. The control Ca 2+ transient (a) was also shown to indicate the rapid Ca 2+ decay (marked by vertical line) on termination of 20 mM K + stimulation. ( E, F ) Superimposition of Ca 2+ transients showing that monensin enhances both nimodipine-sensitive ( E ) and -insensitive ( F ) Ca 2+ transients. ( G ) Normalized Ca 2+ transients to show the effect of nimodipine on the delayed onset of Ca 2+ clearance (marked by horizontal bar). The control Ca 2+ transient was also shown to indicate the rapid Ca 2+ decay (marked by vertical line) on termination of 20 mM K + stimulation.

Article Snippet: Nimodipine was purchased from Tocris Cookson (Ellisville, MO, USA), and ouabain and monensin from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Role of Intracellular Na + in the Regulation of [Ca 2+ ] i in the Rat Suprachiasmatic Nucleus Neurons

doi: 10.3390/ijms20194868

Figure Lengend Snippet: The plateau-like slow decay phase is independent of nimodipine-sensitive Ca 2+ influx. A representative experiment to show the effect of 2 μM nimodipine on the 20 K + -evoked Ca 2+ transient (an average of 12 cells). Note that the plateau-like slow decay phase (marked by arrowhead) still occurred in the presence of nimodipine to block Ca 2+ entry via L-type Ca 2+ channels. Arrows mark the application of 2 µM nimodipine.

Article Snippet: Nimodipine was purchased from Tocris Cookson (Ellisville, MO, USA), and ouabain and monensin from Sigma-Aldrich (St Louis, MO, USA).

Techniques: Blocking Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 1. Nimodipine attenuates retinal inflammation in EAU mice. (A) Representative fundus photos showed that nimodipine decreased retinal inflam- mation, characterized by less vasculitis, fewer linear lesions, and minimal confluent tissue destruction of the retinas (n = 10). (B) On day 21, the eyes were enucleated and prepared for histopathology. Nimodipine treatment reduced the inflamed lesions greatly. The histopathologic scores were statistically different (n = 10). (CE) The intraocular infiltrated cells were isolated and measured by flow cytometry on day 21 after immunization. Nimodipine decreased CD41

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: Histopathology, Isolation, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 2. Nimodipine modulates the systemic immune profiles. (A) Representative flow cytometry analysis of the naive and memory CD41 T cells. Nimodipine inhibited the naive CD41 T cells (CD44CD62L1) differentiation into memory CD41 T cells (CD441CD62L). (B) Nimodipine decreased the levels of the Th1 and Th17 cells of the DLNs. (C) The proportion of Tregs in DLNs were increased more significantly in the nimodipine-treated mice than in the vehicle-treated mice. Flow cytometry experiments above were gated on CD4. Data are representative of at least six independent experiments and pre- sented as the mean ± SD. *p < 0.05, **p < 0.01.

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 3. Nimodipine decreases IRBP120specific immunological response in vitro. The lymphocytes from DLNs of EAU mice were collected and cul- tured in vitro with IRBP120 for 3 d. (A) The percentages of viable cells (zombie) were similar at concentrations of 0, 20, and 40 mM of nimodipine, whereas 80 mM nimodipine treatment decreased the percentage. (BD) Nimodipine (20 and 40 mM) suppressed the differentiation of Th17 and Th1 cells in a dose-dependent manner with no effect on TNF-asecreting cells (n = 7). (E and F) ELISA results showed that nimodipine reduced the releases of the IL-17A and IFN-g in a dose-dependent manner (n = 7). (G) Expression of TNF-a showed no statistical difference between nimodipine and vehicle groups (n = 7). Flow cytometry experiments above were gated on CD4. The values represent the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: In Vitro, Enzyme-linked Immunosorbent Assay, Expressing, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 4. Nimodipine inhibits LPS-stimulated immunological responses in vitro. The PBMCs from uveitis patients were collected and cultured in vitro with LPS (100 ng/ml) for 3 d. (A) The viability of cells was not greatly affected by nimodipine at the concentrations of 0, 10, 20 and 40 mM. However, nimo- dipine (80 mM) decreased viable cells significantly. (B and C) Nimodipine (20 and 40 mM) decreased the expression of IL-17A and IFN-g of CD41 T cells (n = 5). (D) The TNF-a secretion of LPS-stimulated PBMCs was not influenced by nimodipine (n = 5). Flow cytometry experiments above were gated on CD31CD8. The values represent the mean ± SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: In Vitro, Cell Culture, Expressing, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 5. Nimodipine suppresses the energy metabolism, proliferation, activation, and differentiation of T cells. (A) Energy metabolism was evaluated by ROS production. Nimodipine decreased the ROS production of CD41 T cells (n = 6). (B) Nimodipine inhibited the proliferation of CD41 T cells dose dependently (n = 6). (C) Nimodipine (20 and 40 mM) suppressed the early activation of the naive CD41 T cells characterized by the downregulated CD69 expression (cultured for 6 h) (n = 6). (D) The Th1 polarization characterized by IFN-g expression was inhibited by nimodipine (20 and 40 mM) with a statis- tical difference (n = 6). Flow cytometry experiments above were gated on CD4. The values represent the mean ± SD. ****p < 0.0001.

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: Activation Assay, Expressing, Cell Culture, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 6. Nimodipine alleviates EAU through p38MAPK signaling. (A) DEGs were explored and presented in a heatmap of mRNA abundance. Rows denote RNA expression according to their enrichment in EAU vehicle versus EAU 1 nimodipine (n = 3). (B) The bubble plot from the KEGG analysis showed a significant difference in the MAPK signaling pathway between the two groups. (C) Circle plot for GO analysis of pathways that significantly enriched in EAU 1 nimodipine group. The z-score meant the changes in the pathways. The red or blue points referred to the DEGs that contributed to the pathways shown in the right sheet. (D) Heatmap for GSVA analysis that identified the differentially expressed pathways in each sample. (E) The flow cytom- etry analysis showed that the expression of p-p38 in CD41 T cells of DLNs was higher in the EAU vehicle group than the nonimmunization group, and nimodipine treatment significantly downregulated the p-p38 expression (n = 6). (F) Lymphocytes from DLNs of EAU mice were stimulated with IRBP120 for 3 d in the presence of nimodipine or vehicle. Nimodipine (20 and 40 mM) treatment downregulated the p-p38 expression significantly (n = 6). (G) Nimo- dipine suppressed the CD69 expression at 6 h, which could be compromised by asiatic acid (20 mM), a p38 activator (n = 6). Flow cytometry experiments above were gated on CD4. The values represent the mean ± SD. **p < 0.01, ****p < 0.0001.

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: RNA Expression, Expressing, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: The Calcium Channel Inhibitor Nimodipine Shapes the Uveitogenic T Cells and Protects Mice from Experimental Autoimmune Uveitis through the p38-MAPK Signaling Pathway.

doi: 10.4049/jimmunol.2100568

Figure Lengend Snippet: FIGURE 7. The p38MAPK signaling is involved in the inhibitory effect of nimodipine on the pathogenicity of T cell. (A) The fundus photo showed that IRBP120treated T cells induced uveitis successfully, and T cells pretreated with nimodipine failed to induce uveitis. IRBP120treated T cells in the pres- ence of p38 activator asiatic acid (AA) resulted in severe uveitis, characterized by the prominent swelling of retina, tortuous and dilated vessels, and more patchy inflammatory infiltration. Nimodipine only partially blocked the pathogenicity of T cells caused by AA, manifesting as mild uveitis. Clinical scores of the nimodipine group had a significant difference with vehicle group and nimodipine plus AA group (n = 6). (B) On day 14, the eyes were enucleated and prepared for histopathology. The histopathologic scores were statistically different (n = 6). The representative results are presented. The values represent the mean ± SD. *p < 0.05, ****p < 0.0001.

Article Snippet: The cells were seeded into 96-well plates at a density of 2 105 cells per well and cultured in complete RPMI-1640 medium (with 10% FBS, 1% penicillin streptomycin, 1% sodium pyruvate, and 0.1% b-mercaptoethanol) supplemented with 20 mg/ml hIRBP1 20 at 37 C and 5% CO2 and treated with nimodipine (CAS no. 66085-59-4, Selleck Chemicals) at different concentrations (0, 10, 20, 40, or 80 mM).

Techniques: Histopathology

Journal: Frontiers in Pharmacology

Article Title: Therapeutic effects of traditional Chinese medicine Hua-Feng-Dan in a rat model of ischemic stroke involve renormalization of gut microbiota

doi: 10.3389/fphar.2025.1485340

Figure Lengend Snippet: Hua-Feng-Dan ameliorates the size of the brain infarct and the neurological impairments. Animals were subjected to middle cerebral artery occlusion or sham surgery, then left untreated (MCAO, Sham) or treated with nimodipine (NMDP) or Hua-Feng-Dan at a low dose (HFD-L, 0.162 g/kg), intermediate dose (HFD-M, 0.324 g/kg) or high dose (HFD-H, 0.648 g/kg). (A) Representative photographs of ischemic halves of brains after TTC staining. (B) Extent of cerebral infarction (n = 3). (C) Brain water content (n = 4). (D) Neurological score (n = 20). Quantitative data are mean ± SD. *** P < 0.001 vs. Sham group; ## P < 0.01, ### P < 0.001 vs. MCAO group; based on one-way ANOVA and Tukey’s multiple-comparisons post hoc test.

Article Snippet: Rats were randomized to receive, by oral gavage, saline instead of drug treatment or one of the following drugs dissolved in saline: the calcium channel blocker nimodipine (NMDP, batch 211274, Yabao Pharmaceutical, Shanghai, China), which is widely used to ischemic cerebrovascular disease; and Hua-Feng-Dan.

Techniques: Staining

Journal: Frontiers in Pharmacology

Article Title: Therapeutic effects of traditional Chinese medicine Hua-Feng-Dan in a rat model of ischemic stroke involve renormalization of gut microbiota

doi: 10.3389/fphar.2025.1485340

Figure Lengend Snippet: Hua-Feng-Dan alleviates histopathology injury and apoptosis of nerve cells after ischemic stroke in rats. Animals were subjected to middle cerebral artery occlusion or sham surgery, then left untreated (MCAO, Sham) or treated with nimodipine (NMDP) or Hua-Feng-Dan at a low dose (HFD-L, 0.162 g/kg), intermediate dose (HFD-M, 0.324 g/kg) or high dose (HFD-H, 0.648 g/kg). (A) Thin sections of brain tissue after hematoxylin-eosin staining (n = 3). Blue arrows represent macrophage infiltration or vascular stasis; red arrows, necrotic neurons or pyramidal cells; green arrows, glial cell proliferation; yellow arrows, vacuoles or septa; and black arrows, crumpled neurons or pyramidal cells. Scale bar, 100 μm. (B) Thin sections of brain tissue after terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining (n = 3). Scale bar, 100 μm. (C) TUNEL positive cells density (n = 3).

Article Snippet: Rats were randomized to receive, by oral gavage, saline instead of drug treatment or one of the following drugs dissolved in saline: the calcium channel blocker nimodipine (NMDP, batch 211274, Yabao Pharmaceutical, Shanghai, China), which is widely used to ischemic cerebrovascular disease; and Hua-Feng-Dan.

Techniques: Histopathology, Staining, End Labeling, TUNEL Assay

Journal: Frontiers in Pharmacology

Article Title: Therapeutic effects of traditional Chinese medicine Hua-Feng-Dan in a rat model of ischemic stroke involve renormalization of gut microbiota

doi: 10.3389/fphar.2025.1485340

Figure Lengend Snippet: Hua-Feng-Dan restores the integrity of the intestinal barrier after ischemic stroke in rats. Animals were subjected to middle cerebral artery occlusion or sham surgery, then left untreated (MCAO, Sham) or treated with a high dose of Hua-Feng-Dan (HFD, 0.648 g/kg) and nimodipine (NMDP). (A) Representative photographs of colon H&E staining (n = 3). Red arrows indicate the presence of lymphocytic infiltration; blue arrows, edema or edematous mucosal epithelial cells; black arrows, necrosis of mucosal epithelial cells; yellow arrows, dilated intestinal glands; and purple arrows, vasodilation. Scale bar = 200 and 100 μm. (B–D) Comparison of levels of three indices of intestinal barrier permeability: lipopolysaccharide (LPS), diamine oxidase (DAO) and d -lactate ( d -LA). (E–G) Comparison of levels of three pro-inflammatory factors: tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6. (H) Comparison of levels of total cholesterol (T-CHO) as an index of dyslipidemia. (I, J) Comparison of levels of oxidative stress factors total superoxide dismutase (T-SOD) and malondialdehyde (MDA). Data are mean ± SD, n = 6 (excluding H&E staining). * P < 0.05, ** P < 0.01, *** P < 0.001 vs. Sham group; # P < 0.05, ## P < 0.01, ### P < 0.001 vs. MCAO group (based on one-way ANOVA and Tukey’s multiple-comparisons post hoc test).

Article Snippet: Rats were randomized to receive, by oral gavage, saline instead of drug treatment or one of the following drugs dissolved in saline: the calcium channel blocker nimodipine (NMDP, batch 211274, Yabao Pharmaceutical, Shanghai, China), which is widely used to ischemic cerebrovascular disease; and Hua-Feng-Dan.

Techniques: Staining, Comparison, Permeability