Cardiovascular Research

Regulation of arachidonic acid, eicosanoid, and phospholipase A2 levels in murine mast cells by recombinant stem cell factor.

Authors: A N Fonteh|||J M Samet|||F H Chilton

Affiliations: + Expand

Abstract

The current study evaluates the capacity of recombinant rat stem cell factor (rrSCF) to regulate enzymes that control AA release and eicosanoid generation in mouse bone marrow-derived mast cells (BMMCs). Initial studies indicated that rrSCF provided for 24 h inhibited the release of AA into supernatant fluids of antigen- and ionophore A23187-stimulated BMMCs. Agonist-induced increases in cellular levels of AA were also inhibited, albeit to a lesser degree by rrSCF. To determine the inhibitory mechanism, several steps (e.g., mobilization of cytosolic calcium, release of BMMC granules, and regulation of phospholipase A2 [PLA2] activity) that could influence AA release were measured in rrSCF-treated cells. rrSCF did not alter the capacity of BMMCs to mobilize cytosolic calcium or release histamine in response to antigen and ionophore. BMMCs released large amounts of PLA2 with characteristics of the group II family in response to antigen and ionophore A23187. rrSCF treatment of BMMCs reduced the secretion of this PLA2 activity by BMMCs. Partial purification of acid-extractable PLA2 from rrSCF-treated and untreated BMMCs suggested that rrSCF decreased the quantity of acid-stable PLA2 within the cells. In contrast to group II PLA2, the quantity of cPLA2 (as determined by Western blot analysis) increased in response to rrSCF. To assess the ramifications of rrSCF-induced reductions in AA and group II PLA2, eicosanoid formation was measured in antigen- and ionophore-stimulated BMMCs, rrSCF-inhibited (100 ng/ml, 24 h) prostaglandin D2 (PGD2), thromboxane B2, and leukotriene B4 by 48.4 +/- 7.7%, 61.1 +/- 10.0% AND 38.1 +/- 3.6%, respectively, in antigen-stimulated cells. Similar patterns of inhibition were observed in ionophore-stimulated BMMCs. The addition of a group I PLA2 or exogenous AA to BMMCs reversed the inhibition of eicosanoid generation induced by rrSCF. Together, these data indicate that rrSCF differentially regulates group II and cytosolic PLA2 activities in BMMCs. The resultant reductions in eicosanoid generation suggest that group II PLA2 provides a portion of AA that is used for eicosanoid biosynthesis by BMMCs.

---

Chemical List

Eicosanoids|||Fatty Acids, Nonesterified|||Hematopoietic Cell Growth Factors|||Recombinant Proteins|||Stem Cell Factor|||Arachidonic Acid|||Calcimycin|||Phospholipases A|||Phospholipases A2|||Calcium

---

Reference List

J Clin Invest. 1994 Feb;93(2):571-7|||J Biol Chem. 1994 Feb 25;269(8):5897-904|||J Clin Invest. 1994 Mar;93(3):1224-33|||Proc Natl Acad Sci U S A. 1994 Apr 12;91(8):3142-6|||J Clin Invest. 1994 Apr;93(4):1583-91|||Biochem J. 1994 Apr 1;299 ( Pt 1):197-201|||J Biol Chem. 1994 May 6;269(18):13057-60|||J Immunol. 1994 Jun 1;152(11):5438-46|||Br J Cancer. 1994 Jun;69(6):1166-70|||Biochim Biophys Acta. 1994 May 25;1226(2):201-5|||Eur J Pharmacol. 1994 Feb 21;253(1-2):155-61|||J Allergy Clin Immunol. 1994 Jul;94(1):109-19|||Biochem Pharmacol. 1994 Jul 5;48(1):1-10|||J Biol Chem. 1994 Oct 21;269(42):25999-6005|||Biochem Biophys Res Commun. 1994 Sep 30;203(3):1716-24|||Can J Biochem Physiol. 1959 Aug;37(8):911-7|||J Lab Clin Med. 1984 Dec;104(6):873-81|||J Rheumatol. 1985 Apr;12(2):211-6|||Biochemistry. 1986 Dec 30;25(26):8381-5|||Prostaglandins. 1986 Dec;32(6):889-906|||Biochem Biophys Res Commun. 1987 Jun 30;145(3):1126-33|||J Biol Chem. 1988 Apr 25;263(12):5732-8|||Mol Pharmacol. 1988 Sep;34(3):256-64|||Biochim Biophys Acta. 1988 Dec 16;963(3):476-92|||J Biochem. 1988 Nov;104(5):767-72|||J Biol Chem. 1989 Apr 5;264(10):5335-8|||J Biol Chem. 1989 Apr 5;264(10):5768-75|||J Biochem. 1988 Sep;104(3):326-8|||J Biochem. 1989 Apr;105(4):520-5|||J Biol Chem. 1990 Apr 5;265(10):5409-13|||Chem Immunol. 1990;49:173-205|||J Biol Chem. 1990 Aug 25;265(24):14654-61|||Biochem J. 1990 Oct 1;271(1):37-43|||J Immunol. 1991 Jun 1;146(11):3904-10|||FEBS Lett. 1991 May 6;282(2):326-30|||Cell. 1991 Jun 14;65(6):1043-51|||Adv Exp Med Biol. 1990;279:1-22|||J Biol Chem. 1991 Aug 15;266(23):14850-3|||Biochem Biophys Res Commun. 1991 Aug 15;178(3):1298-305|||FEBS Lett. 1991 Dec 9;294(3):247-51|||Eur J Biochem. 1991 Dec 18;202(3):873-80|||J Immunol. 1992 Feb 1;148(3):772-7|||J Exp Med. 1992 Jan 1;175(1):237-44|||Neurochem Res. 1991 Jun;16(6):651-8|||DNA Cell Biol. 1992 Apr;11(3):233-43|||J Biol Chem. 1992 May 5;267(13):8846-51|||J Immunol. 1992 Jul 15;149(2):599-608|||Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6147-51|||Biochim Biophys Acta. 1992 Jul 29;1127(2):163-7|||Eur J Biochem. 1992 Oct 1;209(1):257-65|||J Biol Chem. 1992 Nov 25;267(33):23451-4|||Eur J Biochem. 1992 Nov 15;210(1):169-76|||Blood. 1992 Dec 15;80(12):3009-21|||J Biol Chem. 1993 Jan 15;268(2):839-44|||J Immunol. 1993 Jan 15;150(2):563-70|||J Clin Invest. 1993 Jan;91(1):115-22|||J Biol Chem. 1993 Feb 5;268(4):2865-71|||J Biol Chem. 1993 Mar 5;268(7):4939-44|||Biochim Biophys Acta. 1993 May 20;1168(1):37-44|||J Biol Chem. 1993 Jun 5;268(16):11697-702|||J Immunol. 1993 Jul 1;151(1):359-66|||J Biol Chem. 1993 Aug 5;268(22):16795-802|||Adv Second Messenger Phosphoprotein Res. 1993;28:81-9|||Int Arch Allergy Immunol. 1993;102(2):170-5|||J Biol Chem. 1993 Oct 15;268(29):21875-82|||J Clin Invest. 1993 Oct;92(4):1639-49|||Biochem Biophys Res Commun. 1993 Oct 15;196(1):147-53|||J Clin Invest. 1993 Nov;92(5):2516-23|||Clin Chem. 1993 Dec;39(12):2453-9|||Eur J Biochem. 1993 Dec 15;218(3):807-13|||J Biol Chem. 1994 Jan 28;269(4):2365-8|||J Biol Chem. 1994 Feb 18;269(7):5092-8