Biochemical Aspects of Prostaglandins and Thromboxanes. by Norman Kharasch

By Norman Kharasch

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Extra resources for Biochemical Aspects of Prostaglandins and Thromboxanes. Proceedings of the 1976 Intra-Science Research Foundation Symposium December 1–3, Santa Monica, California

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4 ÷ 10 " cp m pe r dis h o f 1 ÷ 10 6 cells ) wer e treate d wit h 2 ml o f mediu m contain in g th e indicate d substanc e (bovin e albumin , crystallize d 4 mg/ml ; indomethacin , 5 ÷ 10* 6 M; an d bradykinin, 2 yg/ml ) fo r 1 5 min . When indomethaci n wa s used , th e dishe s wer e preincubate d wit h th e dru g fo r 2 0 mi n befor e th e treatmen t describe d above . Aliquots o f th e mediu m wer e analyze d b y thin-laye r chromatography . Duplicat e dishe s wer e use d an d th e valu e obtaine d agree d withi n 10 % of th e mea n value .

Th e specifi c activitie s o f thes e prostaglandi n synthetase s measure d b y th e formatio n o f PGF2a wer e foun d t o b e 3 3 ng/min/m g protei n fo r contro l an d actino myci n D-treate d cell s an d 3 0 ng/min/m g protei n fo r th e cycloheximide-treate d cells . 3 ) Prostaglandi n synthetas e activit y o f microsoma l preparation s fro m MC5- 5 cell s wa s no t inhibite d i n vitr o b y actinomyci n D o r cycloheximid e at concentration s o f 10 0 yg/ml . Indomethaci n di d inhibi t PGF2a formatio n b y 90% .

Simila r studie s wit h indomethaci n indicate d tha t eve n a te n minut e preincubatio n was sufficien t t o caus e complet e inhibitio n o f prostaglandi n production . Al l o f th e steroida l anti-inflammator y drug s teste d inhibite d th e serum-stimulate d productio n o f prostaglandin s by MC5- 5 cell s (11) . Dexamethason e wa s on e o f th e mor e poten t steroid s (Fig . 2 ) . Maximu m inhibitio n (70% ) wa s obtaine d a t 1 0 " M, an d hal f maxima l inhibitio n (ID50 ) 6 Deacylatio n of Phospholipid s 1 0 -· 10" · I0" Dexomtthotont , Ì 7 ßï-· 19 10 Fig .

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