Arachidonic acid-metabolites produced in cells isolated from rat heart

J Mol Cell Cardiol21

(Supplement

IV)(1989)

FC52DIFFERENCES IN THE PRODUCTION OF EICOSANOIDS BY MACRO- AND MICROVASCULAR ENDOTHELIAL CELLS. P.Rosen, S.Zink, R.Oestreich, Diabetes-Forschungsinstitut, Diisseldorf, FRG Since it became obvious that several features distinguish capillary endothelial cells from those of macrovascular origin, we studied the whether also the formation of eicosanoids is different in enquestion dothelial cell (EC) of different vascular origin. Therefore the formation of eicosanoids by bovine aortic EC (BAEC) and bovine retinal EC (BREC) isolated by collagenase digestion and cultivated in DMEM with of eicosanoPGI was the main product fetal calf serum was determined. to about 70% of total id formation by BAEC. With 10 u.N AA SGI amounted eicosanoids whereas other prostaglandinz (PGs) and lipoxygenase products (LOX) were only detected in minor amounts. As compared to BEAC, PG-synthesis by BREC was reduced to about 50% and PGE became the major PG. The rat 3 o PGI /PGE was higher LOX products were not detectable at all. EC than 10 in BAEC and became as low as 0.5 in BREC. Thus 2.micr 2 vacular differ from macrovascular EC by the reduced capacity for the conversion of AA, by the reduced synthesis of eicosanoids after stimulation by and complete absence of LOX. Synthesis of PGE2 A23187 and bradykinin, seems to be characteristic feature of microvascular EC.

FC~~A~AC~~D~NICACID-NETABOLITES

PBDDDCED IN CELL8 ISOLATED FBDN NAT HURT. M.C.J.G. Linssen. P.J.M.R. Lematens. M. van Bilsen. W. Engels and G.J. van der Vusse. Dept. Physiology and Med. Microbiology, Univ. of Limburg. Maastricht, The Netherlands. To investigate which cell types in the heart are capable to produce arachidonate metabolites. myocytes and non-myocytes were isolated from adult rat hearts after perfusion with collagenase. Rod-shaped myocytes were allowed to attach on dishes and used four hours after isolation. The non-myocytes were grown to confluency under tissue culture conditions (primary non-myocytes). In a subset of experiments subcultivations of the cells were used after one or two passages. The cells were stimulated with 10 uH A23187 and 80 pH arachidonic acid in Hanks’ balanced salt solution with Hepes buffer. After 30 q in incubation at 37’C the formed products were measured with high pressure liquid chrollatography and radio-irmuno-assay techniques. Prostaglandin or throtsboxane production by myocytes was less than the detection level of HPLC (< 1 ng/mg protein1 whereas the primary non-myocytes produced amounts of PGEs 1212 + 54 ng/mg protein). L-keto-Fla (a stable degradation product of PGIzl (59 + 15) and TXBl (2,O + 0.5). The production in cultures of non-myocytes after the first and second passage -was found to be 379 + 270. 159 l 87 and < 1 ng/mg protein respectively. Results with the RIA indicated that liitle amounts of 6-keto-Fla (0.5 ng/mg protein) were formed in q yocytes. In conclusion the mass of PC’s and TX (under maximal stimulation) are not formed by cardiomyocytes but by non-myocytes.

FC54REPERFUSION

OF

PRODUCTION. A.G. Semb, J. LJepts. Physiology

HUMAN Vaage-. and

NYOCARDIUH: K. Forsdahl, surgery-,

GRANULOCYTE D. University

TRAPPING

Sorlle., oi

M. Lie*, I‘l-omse,

AND U.D. Norway.

EICOSANOID N)os.

Granulocytes (PIIN) and elcosanolds are possible mediators of reperfuarterial and coronary sinus concentrations 610" 1n,ury. We measured and LTB. of PMN, b-keto-PLF,. (stable metabollte of prostacyclln) during myocardlal reperiuslon I" patients undergolng open heart surgslgnlficant sequestration of PMN (p
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