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Background  Hematopoietic prostaglandin D 2  synthase (H-PGDS, GST Sigma) is a member of the glutathione S-transferase super family of enzymes that catalyses the conjugation of electrophilic substances with reduced glutathione. The enzyme catalyses the conversion of PGH 2  to PGD 2  which mediates inflammatory responses. The inhibition of H-PGDS is of importance in alleviating damage to tissues due to unwarranted synthesis of PGD 2 .  Combretum molle  has been used in African ethno medicinal practices and has been shown to reduce fever and pain. The effect of  C. molle  alkaloid extract on H-PGDS was thus, investigated.    Methods  H-PGDS was expressed in  Escherichia coli  XL1-Blue cells and purified using nickel immobilized metal affinity chromatography. The effect of  C. molle  alkaloid extract on H-PGDS activity was determined with 1-chloro-2, 4-dinitrobenzene (CDNB) as substrate. The effect of  C. molle  alkaloid extract with time on H-PGDS was determined. The mechanism of inhibition was then investigated using CDNB and glutathione (GSH) as substrates.    Results  A specific activity of 24 μmol/mg/min was obtained after H-PGDS had been purified. The alkaloid extract exhibited a 70% inhibition on H-PGDS with an IC 50  of 13.7 μg/ml.  C. molle  alkaloid extract showed an uncompetitive inhibition of H-PGDS with K i  = 41 μg/ml towards GSH, and non-competitive inhibition towards CDNB with K i  = 7.7 μg/ml and K i  ′  = 9.2 μg/ml.    Conclusion  The data shows that  C. molle  alkaloid extract is a potent inhibitor of H-PGDS. This study thus supports the traditional use of the plant for inflammation.

Inhibition of hematopoietic prostaglandin D2 Synthase (H-PGDS) by an alkaloid extract from Combretum molle