Korean J Physiol Pharmacol.  2014 Jun;18(3):241-247. 10.4196/kjpp.2014.18.3.241.

Effects of C18 Fatty Acids on Intracellular Ca2+ Mobilization and Histamine Release in RBL-2H3 Cells

Affiliations
  • 1College of Pharmacy, Chung-Ang University, Seoul 156-756, Korea. simss@cau.ac.kr

Abstract

To investigate the underlying mechanisms of C18 fatty acids (stearic acid, oleic acid, linoleic acid and alpha-linolenic acid) on mast cells, we measured the effect of C18 fatty acids on intracellular Ca2+ mobilization and histamine release in RBL-2H3 mast cells. Stearic acid rapidly increased initial peak of intracellular Ca2+ mobilization, whereas linoleic acid and alpha-linolenic acid gradually increased this mobilization. In the absence of extracellular Ca2+, stearic acid (100 microM) did not cause any increase of intracellular Ca2+ mobilization. Both linoleic acid and alpha-linolenic acid increased intracellular Ca2+ mobilization, but the increase was smaller than that in the presence of extracellular Ca2+. These results suggest that C18 fatty acid-induced intracellular Ca2+ mobilization is mainly dependent on extracellular Ca2+ influx. Verapamil dose-dependently inhibited stearic acid-induced intracellular Ca2+ mobilization, but did not affect both linoleic acid and alpha-linolenic acid-induced intracellular Ca2+ mobilization. These data suggest that the underlying mechanism of stearic acid, linoleic acid and alpha-linolenic acid on intracellular Ca2+ mobilization may differ. Linoleic acid and alpha-linolenic acid significantly increased histamine release. Linoleic acid (C18:2: omega-6)-induced intracellular Ca2+ mobilization and histamine release were more prominent than alpha-linolenic acid (C18:3: omega-3). These data support the view that the intake of more alpha-linolenic acid than linoleic acid is useful in preventing inflammation.

Keyword

Ca2+ mobilization; C18 fatty acids; Histamine release; PLC assay

MeSH Terms

alpha-Linolenic Acid
Fatty Acids*
Histamine Release*
Inflammation
Linoleic Acid
Mast Cells
Oleic Acid
Verapamil
Fatty Acids
Linoleic Acid
Oleic Acid
Verapamil
alpha-Linolenic Acid

Figure

  • Fig. 1 Cytotoxicity of C18 fatty acids in RBL-2H3 cells. Cells were incubated with C18 fatty acids (A: stearic acid, B: oleic acid, C: linoleic acid, D: α-linolenic acid) for 24 h in a 5% CO2 incubator at 37℃. The number of living cells was measured using MTT. Results are mean±S.D. from four separate experiments.

  • Fig. 2 C18 fatty acid-induced intracellular Ca2+ mobilization. Intracellular Ca2+ mobilization was measured in Fura-2/AM-loaded RBL-2H3 cells. Intracellular Ca2+ concentration induced by a 100-sec exposure to the C18 fatty acids [A: C18 fatty acids, B: stearic acid (SA), C: linoleic acid (LA), D: α-linolenic acid (LN)] was expressed as the ratio of F340/F380. Results are the representative data of four separate experiments.

  • Fig. 3 C18 fatty acids-induced intracellular Ca2+ mobilization in the presence or absence of extracellular Ca2+. Intracellular Ca2+ concentration was induced by a 100-sec exposure to the C18 fatty acids [A: stearic acid (SA, 100 µM), B: linoleic acid (LA, 100 µM), C: α-linolenic acid (LN, 100 µM)]. Results are the representative data of four separate experiments.

  • Fig. 4 Effect of verapamil on intracellular Ca2+ mobilization induced by C18 fatty acids. After a 50-sec treatment with 10 µM verapamil (VP), intracellular Ca2+ concentration was induced by a 100-sec exposure to the C18 fatty acids [A: stearic acid (SA), B: linoleic acid (LA), C: α-linolenic acid (LN)]. Results are the representative data of four separate experiments.

  • Fig. 5 (A) Dose-response of verapamil (VP) to intracellular Ca2+ mobilization and (B) peak ratio induced by stearic acid (SA, 100 µM). Results are the representative data of four separate experiments. *Significantly different from control (p<0.05).

  • Fig. 6 Effect of bisindolylmaleimide (BIM) on intracellular Ca2+ mobilization induced by a 100-sec exposure to C18 fatty acids [A: stearic acid (SA), B: linoleic acid (LA), C: α-linolenic acid (LN)]. Results are the representative data of four separate experiments.

  • Fig. 7 C18 Fatty acid-induced histamine release in RBL-2H3 cells. (A) Histamine release induced by arachidonic acid (AA, 100 µM), stearic acid (SA, 100 µM), linoleic acid (LA, 100 µM) and α-linolenic acid (LN, 100 µM). (B) Effect of 10 µM bisindolylmaleimide (BIM) on histamine release induced by linoleic acid (LA, 100 µM) and α-linolenic acid (LN, 100 µM). *Significantly different from Control (p<0.05); aSignificantly different from LA (p<0.05); bSignificantly different from LN (p<0.05).


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