Genistein, a potent inhibitor of secretory phospholipase A2: a new insight in down regulation of inflammation.
OBJECTIVE
Some of the legumes, spices and medicinal herbs rich in genistein are known for their anti-inflammatory properties. Anti-inflammatory property of these herbs is determined by subjecting secretory phospholipase A(2) (sPLA(2)) inhibition, a key enzyme in the inflammatory reactions by genistein.


METHODS
Genistein was assessed for inhibition of sPLA(2) activity using (14)C-oleate radiolabelled Escherichia coli membrane as substrate. The enzyme-inhibitor interaction was established by intrinsic fluorescence and circular dichroism studies. The in vivo anti-inflammatory activity was tested by injecting sPLA(2), Vipera russelli venom phospholipase-V (VRV-PL-V) with different concentrations of genistein in the range of 3-21 muM into intra plantar surface of right hind footpad of mice. Systemic effect was tested by administering the genistein (21 muM) i.p. 30 min before and immediately after sPLA(2) injection.


RESULTS
Genistein inhibited sPLA(2) enzymes of inflammatory exudates (human synovial fluid and human pleural fluid) and snake venoms (VRV-PL-V and Naja naja phospholipase-I) in a concentration dependent manner with IC(50) values ranging from 5.75 to 11.75 muM. Increasing the calcium (Ca(2+)) concentration from 2.5 to 15 mM and substrate concentration up to 120 nM did not alter the level of inhibition. Genistein alters the intrinsic fluorescence intensity and shown apparent shift in far ultra violet-circular dichroism spectra of VRV-PL-V, indicating the direct interaction with enzyme. Genistein also inhibited the VRV-PL-V induced mouse paw oedema in a concentration dependent manner. The genistein at 21 muM concentration administered immediately after the VRV-PL-V injection, effectively neutralized the oedema inducing activity.


CONCLUSIONS
Genistein inhibited sPLA(2) activity of both inflammatory exudates and snake venoms in a concentration dependent manner and sPLA(2) induced mouse paw oedema. The study partially explains the observed anti-inflammatory property of several medicinal herbs which containing genistein.
Dharmappa
Kattepura K
KK
Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysore 570006, India.
Mohamed
Riyaz
R
Shivaprasad
Holenarasipura V
HV
Vishwanath
Bannikuppe Sannanaik
BS
D000818:Animals; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation / Q000187:drug effects / Q000502:physiology*; D019833:Genistein / Q000494:pharmacology* / Q000627:therapeutic use; D006801:Humans; D007249:Inflammation / Q000188:drug therapy / Q000201:enzymology; D018836:Inflammation Mediators / Q000494:pharmacology* / Q000627:therapeutic use; D051379:Mice; D054497:Phospholipases A2, Secretory / Q000037:antagonists & inhibitors* / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology* / Q000627:therapeutic use; D012910:Snake Venoms / Q000037:antagonists & inhibitors / Q000201:enzymology
D018836:Inflammation Mediators; D047428:Protein Kinase Inhibitors; D012910:Snake Venoms; D019833:Genistein; D054497:Phospholipases A2, Secretory