Ionomycin free acid
(Synonyms: 离子霉素,SQ23377) 目录号 : GC15446
A calcium ionophore
Cas No.:56092-81-0
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
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- Purity: >98.00%
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- Datasheet
| Cell experiment [1]: | |
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Cell lines |
NK cell |
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Preparation Method |
Cells were then treated with either 1 µM Ionomycin free acid or DMSO, as vehicle control, and cultured for 16 hours. Control and ionomycin-treated cells were then washed and allowed to rest for 24 hours at 2x106 cells/mL in RPMI 10% FBS. |
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Reaction Conditions |
1 µM Ionomycin free acid for 16 hours |
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Applications |
Human NK cells treated with Ionomycin free acid lose their ability to degranulate and secrete IFN-γ in response to a variety of stimuli, but IL-2 stimulation can compensate these defects. |
| Animal experiment [2]: | |
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Animal models |
Athymic nude mice (Balb/c nu/nu female, 6 to 8 weeks old) |
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Preparation Method |
The effects of intratumoral injection of Ionomycin free acid on the growth of subcutaneous HT1376 tumors established in athymic nude mice were then tested. |
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Dosage form |
Intratumoral injection 100 ug Ionomycin free acid 3 times a week for 4 weeks |
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Applications |
Intratumoral injection of Ionomycin free acid into subcutaneous HT1376 tumors reduced the tumorigenicity in nude mice. |
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References: [1].Romera-CÁrdenas G, Thomas LM, et,al. Ionomycin Treatment Renders NK Cells Hyporesponsive. PLoS One. 2016 Mar 23;11(3):e0150998. doi: 10.1371/journal.pone.0150998. PMID: 27007115; PMCID: PMC4805247. |
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Ionomycin free acid is a selective and potent calcium ion carrier that acts as an active Ca2+ carrier. By stimulating the entry of storage-regulated cations across biofilms, effectively improves Ca2+ influx[9].
Human NK cells treated with Ionomycin free acid lose their ability to degranulate and secrete IFN-γ in response to a variety of stimuli, but IL-2 stimulation can compensate these defects[2]. When tested hypothesis by hyperpolarizing HL-60 cells using Ionomycin free acid before electroporation. Hyperpolarizing cells before electroporation alters the pulsed electric field intensity thresholds for reversible electroporation and IRE, allowing for greater control and selectivity of electroporation outcomes[3]. Ionomycin free acid induces calcium influx into the intracellular region and reactive oxygen species production in N1E-115 cells. Lipid hydroperoxide production was induced in ionomycin-treated N1E-115 cells[5]. Ionomycin free acid, at least in part, exerts its effects via specific binding to a G-protein coupled receptor, thereby evoking downstream cellular events like arachidonate release with subsequent prostaglandin formation[6]. A high concentration of Ionomycin free acid increased the frequency and amplitude of calcium oscillation patterns, affecting the balance of mitochondrial energy metabolism, leading to increased reactive oxygen species (ROS) and decreased ATP[1].
Intratumoral injection of Ionomycin free acid into subcutaneous HT1376 tumors reduced the tumorigenicity in nude mice. Furthermore, these in vivo growth-inhibitory effects of Ionomycin free acid were significantly enhanced by pretreatment with cisplatin[8]. Acetylcholine (ACh) evoked secretion by the calcium ionophore, ionomycin, was studied at frog motor nerve endings. Bath application of Ionomycin free acid stimulated an irreversible increase in the rate of spontaneous, quantal ACh release in the presence of extracellular Ca2+. In contrast, local application of Ionomycin free acid stimulated a rapid, reversible acceleration of spontaneous ACh release[4]. Following stimulation with Ionomycin free acid, PD-1+ICOS+ CD4+ T cells expressed significantly lower IL-17A, but not IFNγ, levels in GF BXD2 mice compared to SPF BXD2 mice[7].
References:
[1]. Chen C, Sun T, et,al. Ionomycin-induced mouse oocyte activation can disrupt preimplantation embryo development through increased reactive oxygen species reaction and DNA damage. Mol Hum Reprod. 2020 Oct 1;26(10):773-783. doi: 10.1093/molehr/gaaa056. PMID: 32697831.
[2]. Romera-Cárdenas G, Thomas LM, et,al. Ionomycin Treatment Renders NK Cells Hyporesponsive. PLoS One. 2016 Mar 23;11(3):e0150998. doi: 10.1371/journal.pone.0150998. PMID: 27007115; PMCID: PMC4805247.
[3]. Aiken EJ, Kilberg BG, et,al. Ionomycin-Induced Changes in Membrane Potential Alter Electroporation Outcomes in HL-60 Cells. Biophys J. 2018 Jun 19;114(12):2875-2886. doi: 10.1016/j.bpj.2018.05.018. PMID: 29925024; PMCID: PMC6026377.
[4]. Hunt JM, Silinsky EM. Ionomycin-induced acetylcholine release and its inhibition by adenosine at frog motor nerve endings. Br J Pharmacol. 1993 Oct;110(2):828-32. doi: 10.1111/j.1476-5381.1993.tb13887.x. PMID: 8242258; PMCID: PMC2175912.
[5]. Nakamura S, Nakanishi A, et,al. Ionomycin-induced calcium influx induces neurite degeneration in mouse neuroblastoma cells: analysis of a time-lapse live cell imaging system. Free Radic Res. 2016;50(11):1214-1225. doi: 10.1080/10715762.2016.1227074. Epub 2016 Sep 29. PMID: 27573976.
[6]. Leis HJ, Windischhofer W. Ionomycin induces prostaglandin E2 formation in murine osteoblastic MC3T3-E1 cells via mechanisms independent of its ionophoric nature. Biochem Cell Biol. 2016 Jun;94(3):236-40. doi: 10.1139/bcb-2015-0148. Epub 2016 Feb 9. PMID: 27065246.
[7]. Hong H, Alduraibi F, et,al. Host Genetics But Not Commensal Microbiota Determines the Initial Development of Systemic Autoimmune Disease in BXD2 Mice. Arthritis Rheumatol. 2022 Apr;74(4):634-640. doi: 10.1002/art.42008. Epub 2022 Feb 10. PMID: 34725967; PMCID: PMC9071869.
[8]. Miyake H, Hara I, et,al. Calcium ionophore, ionomycin inhibits growth of human bladder cancer cells both in vitro and in vivo with alteration of Bcl-2 and Bax expression levels. J Urol. 1999 Sep;162(3 Pt 1):916-21. doi: 10.1097/00005392-199909010-00090. PMID: 10458408.
[9]. Erdahl WL, Chapman CJ, et,al. Ionomycin, a carboxylic acid ionophore, transports Pb(2+) with high selectivity. J Biol Chem. 2000 Mar 10;275(10):7071-9. doi: 10.1074/jbc.275.10.7071. PMID: 10702273.
Ionomycin free acid 是一种选择性的强效钙离子载体，可作为活性 Ca2+ 载体。通过刺激存储调节阳离子进入生物膜，有效提高 Ca2+ 内流[9]。
用离子霉素游离酸处理的人 NK 细胞失去脱颗粒和分泌 IFN-γ 的能力以响应各种刺激，但 IL-2 刺激可以弥补这些缺陷[2]。当通过在电穿孔前使用离子霉素游离酸使 HL-60 细胞超极化来检验假设时。电穿孔前超极化细胞会改变可逆电穿孔和 IRE 的脉冲电场强度阈值，从而更好地控制和选择性电穿孔结果[3]。 Ionomycin free acid 在 N1E-115 细胞中诱导钙流入细胞内区域并产生活性氧。在离子霉素处理的 N1E-115 细胞中诱导脂质氢过氧化物的产生[5]。离子霉素游离酸至少部分通过与 G 蛋白偶联受体特异性结合发挥作用，从而引发下游细胞事件，如花生四烯酸释放以及随后的前列腺素形成[6]。高浓度的离子霉素游离酸增加钙振荡模式的频率和幅度，影响线粒体能量代谢的平衡，导致活性氧（ROS）增加和ATP减少[1]。 p>\n
将离子霉素游离酸瘤内注射到皮下 HT1376 肿瘤中可降低裸鼠的致瘤性。此外，通过顺铂预处理显着增强了离子霉素游离酸的这些体内生长抑制作用[8]。在青蛙运动神经末梢研究了乙酰胆碱 (ACh) 引起的钙离子载体、离子霉素的分泌。在细胞外 Ca2+ 存在的情况下，离子霉素游离酸的浴应用刺激了自发的、量子 ACh 释放速率的不可逆增加。相比之下，局部应用离子霉素游离酸可刺激 ACh 自发释放的快速、可逆加速[4]。与 SPF BXD2 小鼠相比，用离子霉素游离酸刺激后，PD-1+ICOS+ CD4+ T 细胞表达的 IL-17A 水平显着低于 SPF BXD2 小鼠[7]。
| Cas No. | 56092-81-0 | SDF | |
| 别名 | 离子霉素,SQ23377 | ||
| 化学名 | (4R,6R,8R,10E,12S,14S,16E,18S,19S,20R,21S)-11,19,21-trihydroxy-22-((2R,2'R,5R,5'R)-5'-((R)-1-hydroxyethyl)-2,5'-dimethyloctahydro-[2,2'-bifuran]-5-yl)-4,6,8,12,14,18,20-heptamethyl-9-oxodocosa-10,16-dienoic acid | ||
| Canonical SMILES | O[C@H](C)[C@]1(C)O[C@H](CC1)[C@]2(C)O[C@@H](C[C@@H]([C@@H](C)[C@H]([C@H](/C=C/C[C@H](C)C[C@H](C)/C(O)=C\C([C@H](C)C[C@H](C)C[C@H](C)CCC(O)=O)=O)C)O)O)CC2 | ||
| 分子式 | C41H72O9 | 分子量 | 709.01 |
| 溶解度 | 1.4mg/mL in DMSO, 2.5mg/mL in DMF | 储存条件 | Desiccate at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.4104 mL | 7.0521 mL | 14.1042 mL |
| 5 mM | 0.2821 mL | 1.4104 mL | 2.8208 mL |
| 10 mM | 0.141 mL | 0.7052 mL | 1.4104 mL |
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动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
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2.
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Liberation of [3H]arachidonic acid and changes in cytosolic free calcium in fura-2-loaded human platelets stimulated by Ionomycin and collagen
Biochem J1986 May 1;235(3):869-77.PMID: 3092807DOI: 10.1042/bj2350869
Cytosolic Ca2+ levels and arachidonate liberation were investigated in platelets loaded with the fluorescent Ca2+ indicator dye fura-2, and labelled with [3H]arachidonate. Fura-2 was used in preference to quin2 because the latter interfered with [3H]arachidonate labelling of phospholipids. From a resting free Ca2+ level of around 100 nM, Ionomycin (10-200 nM) evoked an instantaneous, concentration-dependent increase in cytosolic Ca2+ that only resulted in [3H]arachidonate liberation (up to 4-fold over control) at Ca2+ levels greater than 1 microM. Addition of collagen (10 micrograms/ml) evoked an elevation in Ca2+ up to 461 +/- 133 nM. These changes in Ca2+ were accompanied by a 2-4-fold elevation in [3H]arachidonate with depletion of [3H]phosphatidylcholine by 17 +/- 4% and [3H]phosphatidylinositol by 41 +/- 7%. Indomethacin (10 microM) reduced the elevation in Ca2+ by collagen to 115 +/- 18 nM but did not significantly inhibit the 2-4-fold increase in [3H]arachidonate. [3H]Phosphatidylcholine and [3H]phosphatidylinositol were decreased by 9 +/- 7% and 10 +/- 6%, respectively, with collagen in the presence of indomethacin. Stimulation of phosphoinositide turnover by collagen in the presence and absence of indomethacin was indicated by [32P]phosphatidate formation in cells prelabelled with [32P]Pi. This phosphatidate formation was decreased (75%) by the presence of indomethacin. In the presence of indomethacin, phorbol myristate acetate (20 nM) alone or in combination with Ionomycin (30 nM) failed to stimulate arachidonate liberation despite a marked stimulation of aggregation. These results indicate that, whereas Ionomycin requires Ca2+ in the microM range for arachidonate liberation, collagen, notably in the presence of indomethacin, does so at basal Ca2+ levels. The mechanisms underlying the regulation of arachidonate release by collagen are not clear, but do not appear to involve activation of protein kinase C, or an elevation of cytosolic free Ca2+.
Ionomycin-stimulated arachidonic acid release in human platelets: a role for protein kinase C and tyrosine phosphorylation
Thromb Haemost1996 Aug;76(2):248-52.PMID: 8865540DOI: 10.1002/(SICI)1097-4644(19961201)63:3%3C292::AID-JCB4%3E3.0.CO;2-S
Collagen (10-90 micrograms/ml) and Ionomycin (1 microM; a calcium ionophore) each evoked rises in intracellular free calcium, protein kinase C activity and arachidonic acid release in human platelets, and as previously demonstrated for collagen, Ionomycin (1 microM) stimulated protein tyrosine phosphorylation. However, at lower concentrations (60 and 250 nM) Ionomycin selectively mobilised calcium. Ro31-8220 (a selective inhibitor of protein kinase C) inhibited (by 50%) Ionomycin-stimulated arachidonic acid release. Genistein (an inhibitor of protein tyrosine kinases) also reduced by 50% Ionomycin-stimulated arachidonic acid release. In combination, genistein and Ro31-8220 abolished Ionomycin-stimulated arachidonic acid release. These findings show 1) that a rise in calcium is not sufficient, and 2) the activation of both protein kinase C and protein tyrosine phosphorylation is necessary, for full Ionomycin-stimulated arachidonic acid release in human platelets.
Calcium-calmodulin plays a major role in bradykinin-induced arachidonic acid release by bovine aortic endothelial cells
J Cell Biochem1996 Dec 1;63(3):292-301.PMID: 8913880DOI: 10.1002/(SICI)1097-4644(19961201)63:3%3C292::AID-JCB4%3E3.0.CO;2-S
We provided evidence that calcium-calmodulin plays a major role in bradykinin-induced arachidonic acid release by bovine aortic endothelial cells. In cells labeled for 16 hr with 3H-arachidonic acid, Ionomycin and Ca2(+)-mobilizing hormones such as bradykinin, thrombin and platelet activating factor induced arachidonic acid release. However, arachidonic acid release was not induced by agents known to increase cyclic AMP (forskolin, isoproterenol) or cyclic GMP (sodium nitroprusside). Bradykinin induced the release of arachidonic acid in a dose-dependent manner (EC50 = 1.6 +/- 0.7 nM). This increase was rapid, reaching a maximal value of fourfold above basal level in 15 min. In a Ca2(+)-free medium, bradykinin was still able to release arachidonic acid but with a lower efficiency. Quinacrine (300 microM), a blocker of PLA2, completely inhibited bradykinin-induced arachidonic acid release. The B2 bradykinin receptor antagonist HOE-140 completely inhibited bradykinin-induced arachidonic acid release. The B1-selective agonist DesArg9-bradykinin was inactive and the B1-selective antagonist [Leu8] DesArg9-bradykinin had no significant effect on bradykinin-induced arachidonic acid release. The phospholipase C inhibitor U-73122 (100 microM) decreased bradykinin-induced arachidonic acid release. The calmodulin inhibitor W-7 (50 microM) drastically reduced the bradykinin- and Ionomycin-induced arachidonic acid release. Also, forskolin decreased bradykinin-induced arachidonic acid release. These results suggest that the activation of PLA2 by bradykinin in BAEC is a direct consequence of phospholipase C activation. Ca2(+)-calmodulin appears to be the prominent activator of PLA2 in this system.
L-Ascorbic acid potentiates nitric oxide synthesis in endothelial cells
J Biol Chem1999 Mar 19;274(12):8254-60.PMID: 10075731DOI: 10.1074/jbc.274.12.8254
Ascorbic acid has been shown to enhance impaired endothelium-dependent vasodilation in patients with atherosclerosis by a mechanism that is thought to involve protection of nitric oxide (NO) from inactivation by free oxygen radicals. The present study in human endothelial cells from umbilical veins and coronary arteries investigates whether L-ascorbic acid additionally affects cellular NO synthesis. Endothelial cells were incubated for 24 h with 0.1-100 microM ascorbic acid and were subsequently stimulated for 15 min with Ionomycin (2 microM) or thrombin (1 unit/ml) in the absence of extracellular ascorbate. Ascorbate pretreatment led to a 3-fold increase of the cellular production of NO measured as the formation of its co-product citrulline and as the accumulation of its effector molecule cGMP. The effect was saturated at 100 microM and followed a similar kinetics as seen for the uptake of ascorbate into the cells. The investigation of the precursor molecule L-gulonolactone and of different ascorbic acid derivatives suggests that the enediol structure of ascorbate is essential for its effect on NO synthesis. Ascorbic acid did not induce the expression of the NO synthase (NOS) protein nor enhance the uptake of the NOS substrate L-arginine into endothelial cells. The ascorbic acid effect was minimal when the citrulline formation was measured in cell lysates from ascorbate-pretreated cells in the presence of known cofactors for NOS activity. However, when the cofactor tetrahydrobiopterin was omitted from the assay, a similar potentiating effect of ascorbate pretreatment as seen in intact cells was demonstrated, suggesting that ascorbic acid may either enhance the availability of tetrahydrobiopterin or increase its affinity for the endothelial NOS. Our data suggest that intracellular ascorbic acid enhances NO synthesis in endothelial cells and that this may explain, in part, the beneficial vascular effects of ascorbic acid.
Phosphorylation of cytosolic phospholipase A2 by IL-3 is associated with increased free arachidonic acid generation and leukotriene C4 release in human basophils
J Allergy Clin Immunol1998 Sep;102(3):512-20.PMID: 9768595DOI: 10.1016/s0091-6749(98)70142-3
Background: Human basophils secrete leukotriene C4 (LTC4) in response to various stimuli, and a short treatment with IL-3 enhances LTC4 release, although IL-3 alone does not induce LTC4 release. However, the mechanism of this priming effect of IL-3 for LTC4 generation remains unknown in human basophils.
Objective: This study was designed to explore the mechanisms by which short treatments with IL-3 enhance stimulated secretion of LTC4, with a focus on the activation of cytosolic phospholipase A2 (cPLA2).
Methods: The phosphorylation state of cPLA2 in human basophils was examined by its shift in electrophoretic mobility as detected by Western blotting. Free arachidonic acid (AA) and LTC4 were measured by gas chromatography-negative ion chemical ionization mass spectrometry and LTC4-specific RIA, respectively.
Result: Human basophils expressed cPLA2. IL-3, as well as the protein kinase C (PKC) activator phorbol 12-myristate 13-acetate, caused a shift in the electrophoretic mobility of cPLA2, which indicated phosphorylation of cPLA2 and therefore its activation. Ionomycin at a concentration of 0.1 microg/mL was used to induce a modest elevation of cytosolic calcium response ([Ca2+]I), no apparent cPLA2 phosphorylation, and little free AA and LTC4 generation. Pretreatment with IL-3 (1 to 10 ng/mL) markedly enhanced Ionomycin (0.1 microg/mL)-mediated AA and LTC4 generation. The concentration dependence of cPLA2 phosphorylation by IL-3 and its effects on free AA and LTC4 generation were similar. The selective PKC inhibitors, bis-indolylmaleimide II and Ro-31-8220 inhibited the phorbol 12-myristate 13-acetate-mediated cPLA2 electrophoretic mobility shift, but not the IL-3-mediated shift, suggesting that the IL-3 effect is PKC independent. Both the anaphylatoxin split product of the C component C5 (C5a) and f-Met-Leu-Phe induced PKC-independent cPLA2 phosphorylation with a similar time course most notable for the absence of observable changes in cPLA2 phosphorylation before 30 seconds. These results suggested an explanation for the absence of free AA generation by C5a. When [Ca2+]I was elevated in response to C5a, there was no phosphorylation of cPLA2, and by the time cPLA2 became phosphorylated, [Ca2+]I had returned to resting levels. Treatment with IL-3 preconditioned the cPLA2 by causing its phosphorylation so that the transient [Ca2+]I response, which followed stimulation by C5a, could induce the generation of free AA and LTC4.
Conclusion: Taken together, these results suggest that the effect of IL-3 for free AA generation and LTC4 release might be due to induction of cPLA2 phosphorylation. The studies demonstrated a need for synchronous cPLA2 phosphorylation and elevations in [Ca2+]I.