title: The anti-inflammatory effect of the SOCC blocker SK&F 96365 on mouse lymphocytes after stimulation by Con A or PMA/ionomycin.
abstract: SK&F 96365, 51-(beta-[3-(p-methoxyphenyl)-propyloxy]-p-methoxyphenethyl)-1H-imidazole hydrochloride, has emerged as a useful pharmacological tool in the study of store-operated Ca²⁺ entry (SOCE). But the precise molecular mechanism and effect of SK&F 96365 on mouse lymphocytes are still not well determined. This study investigated the pharmacological profile of SK&F 96365 on mouse lymphocytes stimulated by mitogen concanavalin A (Con A) or by a combination of a protein kinase C (PKC) activator, phorbol 12-myristate 13-acetate (PMA) and a calcium ionophore, ionomycin in vitro. Our results showed that SK&F 96365 pre-treatment diminished the cytosolic calcium rise on lymphocytes induced by ionomycin, PMA/ionomycin, and thapsigargin (TG), respectively. CFDA-SE staining results showed that SK&F 96365 (5-20 μM) inhibited both Con A- and PMA/ionomycin-induced lymphocytes proliferation in a time- and dose-dependent manner. Upon the same stimulation, SK&F 96365 inhibited the expression of CD69 and CD25 on CD3⁺ T lymphocytes in a dose-dependent manner. The cell cycle analyzing results showed that SK&F 96365 caused a G0/G1 phase cell cycle arrest on both Con A- and PMA/ionomycin-activated lymphocytes in a dose-dependent manner. In addition, SK&F 96365 induced a decrease in mitochondrial membrane potential (ΔΨm) and promoted mitochondrial permeability transition (MPT) in both Con A- and PMA/ionomycin-activated lymphocytes. Furthermore, SK&F 96365 significantly inhibited the production of proinflammatory cytokines (interferon (IFN)-γ and tumor necrosis factor (TNF)), and the anti-inflammatory cytokine (IL-10) on both Con A- and PMA/ionomycin-activated lymphocytes. SK&F 96365 did not induce a statistically significant increase in levels of proinflammatory IL-6 and monocyte chemoattractant protein-1 (MCP-1) but of IL-12p70 upon the stimulation of Con A, whereas these three cytokines were markedly inhibited by it upon the stimulation of PMA/ionomycin. This finding revealed that SK&F 96365 exhibited an anti-inflammatory effect on mouse lymphocytes both upon the stimulation of Con A and PMA/ionomycin, and the precise mechanism of SK&F 96365 inhibiting Con A-activated lymphocytes proliferation is different from PMA/ionomycin.
authors.affiliation: Institute of Tissue Transplantation and Immunology, Jinan University, Guangzhou 510632, PR China.
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Ye
authors.forename: Yaxing
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Lu
authors.forename: Xiuyan
authors.initials: X
authors.lastname: Huang
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: Zeng
authors.forename: Xinqiang
authors.initials: X
authors.lastname: Lai
authors.forename: Yaoying
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000276:immunology / Q000494:pharmacology; D015703:Antigens, CD / Q000096:biosynthesis / Q000276:immunology; D002118:Calcium / Q000276:immunology / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000276:immunology / Q000494:pharmacology; D015220:Calcium Channels / Q000276:immunology / Q000378:metabolism*; D002453:Cell Cycle / Q000187:drug effects* / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003208:Concanavalin A / Q000276:immunology / Q000494:pharmacology; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D004305:Dose-Response Relationship, Drug; D005260:Female; D005452:Fluoresceins / Q000032:analysis; D005456:Fluorescent Dyes / Q000032:analysis; D007093:Imidazoles / Q000276:immunology / Q000494:pharmacology*; D007249:Inflammation / Q000188:drug therapy / Q000276:immunology* / Q000473:pathology; D015759:Ionomycin / Q000276:immunology / Q000494:pharmacology; D008213:Lymphocyte Activation / Q000187:drug effects; D018655:Lymphocyte Count; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008928:Mitochondria / Q000187:drug effects / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects* / Q000276:immunology; D013388:Succinimides / Q000032:analysis; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology; D013755:Tetradecanoylphorbol Acetate / Q000276:immunology / Q000494:pharmacology; D019284:Thapsigargin / Q000276:immunology / Q000494:pharmacology
chemical_list: C087165:5-(6)-carboxyfluorescein diacetate succinimidyl ester; D000893:Anti-Inflammatory Agents; D015703:Antigens, CD; D002121:Calcium Channel Blockers; D015220:Calcium Channels; D016207:Cytokines; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007093:Imidazoles; D013388:Succinimides; D003208:Concanavalin A; D015759:Ionomycin; D019284:Thapsigargin; C063159:1-(2-(3-(4-methoxyphenyl)propoxy)-4-methoxyphenylethyl)-1H-imidazole; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium