Nicardipine

Manufactured by Merck Group

Sourced in United States, Australia, United Kingdom, Israel, Sao Tome and Principe

Nicardipine is a laboratory product manufactured by Merck Group. It is a calcium channel blocker used for research purposes.

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52 protocols using nicardipine

Cisplatin Chemosensitivity Assay in NCI-H446 Cells

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To assess the chemosensitivity of tumor cells, cell viability was measured via an MTT assay (Promega Corporation). Cell suspensions were cultured in RPMI-1640 medium (Thermo Fisher Scientific, Inc.) in 96-well flat-bottomed microtiter plates at a density of 1×10⁴ cells/well and incubated overnight at 37°C. For the nicardipine combination experiment, the concentration of nicardipine (Sigma-Aldrich; Merck KGaG) in the culture medium was adjusted to 10 µM and incubated at 37°C for 15 min in advance.
The following concentrations of cisplatin (Sigma-Aldrich; Merck KGaG) were used: 10⁻⁷ M, 10⁻⁶ M, 10⁻⁵ M, 10⁻⁴ M and 10⁻³ M. All experiments were performed in triplicate, for each concentration. NCI-H446 cells cultured in RPMI-1640 medium were used as the blank controls. MTT solution was added (1 mg/ml per well) and incubated at 37°C for 48 h. Cell viability was subsequently analyzed at a wavelength of 590 nm, using a spectrophotometric microplate reader (Bio-Rad Laboratories, Inc.). The response percentage of cell viability to different drug concentrations was calculated as the inhibition rate (average absorbance of treatment wells/control wells ×100%).

Wu Y., Tang H., Si R., Xia S., Wang R, & Wang Q. (2020). MICA enhances sensitivity to cisplatin in patients with extensive small cell lung cancer via downregulation of ABCG2. Oncology Letters, 20(2), 1143-1152.

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Gastrointestinal Tract Length Measurement

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The entire gastrointestinal tract was carefully separated from the mesentery and removed from culled adult male WT and Nlgn3^R451C mice. Using dissecting scissors, the small intestine from the pylorus to the ileocaecal junction was detached from the stomach and caecum, laid onto a flat surface and measured using a tape measure. A digital image of the tissue was then acquired. To assess the maximal length of the tissue, a proportion of the small intestinal gut preparations were then placed in a glass beaker containing the L-type calcium channel blocker, nicardipine (Sigma-Aldrich, Castle Hill, NSW, Australia) (3 mM) for 20 min. The small intestinal preparations were then removed from the nicardipine solution, re-measured and re-imaged for comparison.
Colons (proximal colon to rectum) from WT and Nlgn3^R451C mice were dissected, and the surrounding mesentery was removed. Colon preparations were attached to an organ bath using a small insect pin (Australia Entomology supplies, AUS). Tissue preparations were positioned adjacent to a printed ruler located in the base of the organ bath and a digital image was acquired. Colon lengths were measured using ImageJ Fiji software v2.9.0 and were calibrated according to the ruler measurements.

Hosie S., Abo-Shaban T., Mou K., Balasuriya G.K., Mohsenipour M., Alamoudi M.U., Filippone R.T., Belz G.T., Franks A.E., Bornstein J.C., Nurgali K, & Hill-Yardin E.L. (2024). Faster Gastrointestinal Transit, Reduced Small Intestinal Smooth Muscle Tone and Dysmotility in the Nlgn3R451C Mouse Model of Autism. International Journal of Molecular Sciences, 25(2), 832.

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Screening FDA/EMA Approved Compounds

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The Prestwick chemical library® (1280 FDA/EMA approved compounds, 10 µM in 96-well plates) was purchased from Prestwick chemicals. Avermectin B1a, bepridil, fluticasone proprionate, gliquidone, triclosan and nicardipine were purchased from Sigma-Aldrich.

van de Wiel S.M., Bijsmans I.T., van Mil S.W, & van de Graaf S.F. (2019). Identification of FDA-approved drugs targeting the Farnesoid X Receptor. Scientific Reports, 9, 2193.

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Perfusion Methodology for Tissue Imaging

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Tissues used for imaging and electrophysiological experiments were perfused with KRB solution containing (mM): NaCl, 120.35; KCl, 5.9; NaHCO₃, 15.5; NaH₂PO₄, 1.2; MgCl₂, 1.2; CaCl₂, 2.5; and glucose, 11.5. KRB solution was oxygenated with a mixture of 97% O₂/3% CO₂ and warmed to 37°C. For experiments using 0 mM [Ca²⁺]_o, CaCl₂ was excluded from the KRB solution and 0.5 mM EGTA was added, resulting in a Ca²⁺ free KRB solution. Ca²⁺ free Hanks’ solution contained the following (mM): NaCl, 125; KCl, 5.36; Glucose, 10; Sucrose, 2.9; NaHCO₃, 15.5; KH₂PO₄, 0.44; Na₂HPO₄, 0.33; Hepes, 10. The pH was adjusted to 7.4 with NaOH.
Nifedipine, nicardipine, atropine, guanethidine, L-NNA, CPA, tetracaine, caffeine, EGTA, and GSK 7975A were purchased from Sigma-Aldrich (St. Louis, MO, USA). Ani9 and MRS2500 was purchased from Tocris Bioscience (Ellisville, Missouri, USA). TTX was purchased from Abcam (Cambridge, MA, USA). Atropine, guanethidine, L-NNA, MRS2500, TTX, CPA, tetracaine and caffeine were diluted in deionized water. Nifedipine and nicardipine were diluted in ethanol. Ani9 was diluted in DMSO. All stock solutions of drugs were diluted to the desired final concentrations with KRB solution.

Drumm B.T., Hannigan K.I., Lee J.Y., Rembetski B.E., Baker S.A., Koh S.D., Cobine C.A, & Sanders K.M. (2022). Ca2+ signalling in interstitial cells of Cajal contributes to generation and maintenance of tone in mouse and monkey lower esophageal sphincters. The Journal of physiology, 600(11), 2613-2636.

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Whole-Mount Preparation of Distal Colon

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Sections of the distal colon (1 cm from the rectum) were isolated and immersed for 15 min in phosphate buffered saline (PBS) pH 7.2 containing 1 µM nicardipine as a muscle relaxant (Sigma-Aldrich, UK). They were then cut along the mesenteric line and washed in PBS to remove luminal contents. Subsequently, they were stretched on a wax support with the mucosal layer downwards and fixed for 6/7 h in Zamboni’s fixative (2% paraformaldehyde containing 0.2% picric acid in 0.1 M PBS) at 4 °C. Following fixation, the tissues were washed in dimethylsulfoxide (DMSO) followed by three 10 min washes in PBS and then stored at 4 °C in PBS containing 0.1% sodium azide. Stretched specimens were processed as longitudinal muscle-myenteric plexus whole-mount preparations (LMMPs) by dissociating the different intestinal layers (i.e. mucosa, submucosa and circular muscle) as previously described^{21 (link)}.

Coletto E., Tough I.R., Pritchard S., Hikima A., Jackson M.J., Jenner P., Ray Chaudhuri K., Cox H.M., Iravani M.M, & Rose S. (2021). Dysregulation of epithelial ion transport and neurochemical changes in the colon of a parkinsonian primate. NPJ Parkinson's Disease, 7, 9.

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Yeast Membrane Protein Expression

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Yeast growth media was obtained from Formedium (Hunstanton, Norfolk, UK). The reagents for protein expression, buffer components, CPM dye, verapamil, nicardipine and disodium ATP were from Sigma-Aldrich (Dorset, UK). N- dodecyl- β-D- maltoside (DDM) was from Merck Chemicals (Nottingham, UK). The protein purification columns were from GE healthcare (Buckinghamshire, UK). Brain polar lipids were from Avanti Polar Lipids (Alabaster, Alabama, USA). Hoechst 33342 was from Thermo Fisher scientific (Waltham, Massachusetts, USA). Ivacaftor was from Vertex pharmaceuticals (Boston, Massachusetts, USA). Tariquidar was from BioVision (Cambridge, UK).

Lingam S., Thonghin N, & Ford R.C. (2017). Investigation of the effects of the CFTR potentiator ivacaftor on human P-glycoprotein (ABCB1). Scientific Reports, 7, 17481.

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Nicardipine Acquisition and Characterization

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Nicardipine was purchased from Sigma Aldrich (St. Louis, MO).

Ekins S., Puhl A.C, & Davidow A. (2020). Repurposing the Dihydropyridine Calcium Channel Inhibitor Nicardipine as a Nav1.8 inhibitor in vivo for Pitt Hopkins Syndrome. Pharmaceutical research, 37(7), 127.

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Local and Whole-Embryo Chemical Treatments

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For local treatment with chemicals or proteins, AG1X2-formate beads (for chemicals) or Affigel Blue beads (BIO-RAD, 1537302; for BMP4) were soaked in different concentrations of the desired protein or chemical overnight at 4 °C. Beads were washed in PCS before grafting. Dimethyl sulfoxide (DMSO, 0.2%) or BSA (0.1%) was used to dilute the chemicals or proteins, respectively, and for soaking the control beads. Final concentrations used for microbead-soaking: 50 ng/μl recombinant human BMP4 (R&D systems, 312-BP), 200 µM dorsomorphin dihydrochloride (Tocris, 3093), 2 µM ionomycin (Sigma, I9657). For chemical treatment to the whole embryo, the chemical was diluted first in PBS (1:10 v:v) and then in egg albumen (9:10 v:v), which was used to culture the embryos (under the vitelline membrane). For treatments with VIVIT (Tocris, 3930) and BAY 11-7821 (Tocris, 1744), embryos were first soaked in the chemical diluted in PCS for 1 h, prior to culture with albumen containing the same concentration of the chemical. Final concentrations used for treatment of whole embryos: 20 µM dorsomorphin, 200 µM flufenamic acid (Sigma, F9005), 2 µM ionomycin, 50 µM, nicardipine (Sigma, N7510), 20 µM U73122 (Sigma, U6756), 20 µM U73343 (Sigma, U6881), 12 µM VIVIT (Tocris, 3930), 12.5 µM BAY 11-7821 (Tocris, 1744).

Lee H.C., Oliveira N.M., Hastings C., Baillie-Benson P., Moverley A.A., Lu H.C., Zheng Y., Wilby E.L., Weil T.T., Page K.M., Fu J., Moris N, & Stern C.D. (2024). Regulation of long-range BMP gradients and embryonic polarity by propagation of local calcium-firing activity. Nature Communications, 15, 1463.

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Regulation of Insulin Receptor Expression by D4 Receptor

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To determine the second messenger(s) involved in the regulation of D₄ on insulin receptor expression in WKY cells, several inhibitors or agonists were used: protein kinase C (PKC) inhibitor (PKC inhibitor 19–31, 10⁻⁶ mol/L)23 protein kinase A (PKA) inhibitor (PKA inhibitor 14–22, 10⁻⁶ mol/L)24 PKC activator (PMA, 10⁻⁷ mol/L)25 PKA activator (Sp‐cAMPs, 10⁻⁷ mol/L)26 calcium‐channel blocker (nicardipine, 10⁻⁶ mol/L)21 and calcium‐channel agonist (BAY‐K8644, 10⁻⁶ mol/L).27 Those reagents were added into the cell incubation medium 15 minutes before PD168077 treatment.
PKC inhibitor 19 to 31, PMA, Sp‐cAMPs, nicardipine, and BAY‐K8644 were purchased from Sigma‐Aldrich. PKA inhibitor 14 to 22 was purchased from Calbiochem Company (Darmstadt, Germany).

Zhang Y., Ren H., Lu X., He D., Han Y., Wang H., Zeng C, & Shi W. (2016). Inhibition of D4 Dopamine Receptors on Insulin Receptor Expression and Effect in Renal Proximal Tubule Cells. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 5(4), e002448.

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Pharmacological Screening of Herbal Compounds

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A68930 hydrochloride, (R)-propylnorapomorphine hydrochloride, and cabergoline were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). D-APV, FK506 monohydrate, actinomycin D, L-(−)-norepinephrine (+)-bitartrate salt monohydrate, serotonin, nicardipine, U0126, KN93, oleoyl-L-α-lysophosphatidic acid sodium salt, and Ki 16425 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Neurotransmitter libraries (dopaminergic, adrenergic, serotonergic, cholinergic, histaminergic, metabotropic glutamatergic, and GABAergic) were purchased from Enzo Life Sciences, Inc. (Farmingdale, NY, USA). Ginsenoside kit (Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, and Rg2), used in Fig. 5a, was purchased from Extrasynthese (Genay Cedex, France). Ginsenoside Rb1, Rc, Rd, Re, and Rg1, used in Fig. 5d, were purchased from LKT Laboratories, Inc. (St. Paul, MN, USA). Herbal extract and herbal medicine-derived compound libraries were kindly donated by the Institute of Natural Medicine, University of Toyama (Toyama, Japan). Each herbal extract used in this study was obtained using a standard method described as follows: each herbal medicine (purchased from Tochimoto Tenkaido (Osaka, Japan)) was extracted in water (10-times volume of herbal medicine) at 100 °C for 50 min, evaporated under reduced pressure, and freeze‐dried to obtain a powder extract. Each herbal extract was redissolved in water.

Fukuchi M., Okuno Y., Nakayama H., Nakano A., Mori H., Mitazaki S., Nakano Y., Toume K., Jo M., Takasaki I., Watanabe K., Shibahara N., Komatsu K., Tabuchi A, & Tsuda M. (2019). Screening inducers of neuronal BDNF gene transcription using primary cortical cell cultures from BDNF-luciferase transgenic mice. Scientific Reports, 9, 11833.

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