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Multi-constituent synergism is responsible for anti-inflammatory effect of Azadirachta indica leaf extract.

AbstractCONTEXT:
Azadirachta indica A. Juss. (Meliaceaes) leaves have been used traditionally to treat swelling and rheumatism in Indian cultures.
OBJECTIVE:
To fractionate A. indica leaf extracts using bioactivity guided manner for identification of the active anti-inflammatory principles.
MATERIALS AND METHODS:
Polarity-gradient sequential extracts (petroleum ether, chloroform, methanol, and water) of A. indica leaves were screened for their anti-inflammatory potential using the carrageenan-induced rat paw edema model (1 g/kg). The chloroform extract was sequentially fractionated to obtain n-hexane (F-1), n-hexane-chloroform (F-2), and chloroform (F-3) fractions and their inhibitory effect on rat paw edema was evaluated (500 mg/kg). Inhibitory effect of F-2 on granuloma formation, plasma interleukin (IL-1), and tumor necrosis factor (TNF-α) was assessed at the doses of 100, 200, and 400 mg/kg using the cotton pellet assay in rats. Three sub-fractions (SF-1, SF-2, and SF-3) were obtained upon chromatography of F-2, and their inhibitory effect on cyclooxygenase was assessed at 200 µg/mL concentration. The sub-fractions were subjected to gas chromatography-mass spectrometry (GC-MS).
RESULTS:
All the extracts showed significant anti-inflammatory effect; however, chloroform extract was the most effective against paw edema (53.25% inhibition). The three fractions of chloroform extract showed significant effect, while F-2 being the most potent (51.02%). F-2 demonstrated dose-dependent inhibition of granuloma and cytokines. Interestingly, all the sub-fractions of F-2 inhibited COX-1 and COX-2 with almost equal potential. GC-MS revealed that chemically the sub-fractions were totally different from each other.
DISCUSSION AND CONCLUSION:
Anti-inflammatory effect of A. indica is a result of cumulative and synergistic effects of diversified constituents with varying polarities that collectively exert the effect via suppression of cyclo-oxygenases and cytokines (IL-1 and TNF-α).
AuthorsMuhammad Ihtisham Umar, Mohd Zaini Asmawi, Amirin Sadikun, A M S Abdul Majid, Item Justin Atangwho, Mohamed B Khadeer Ahamed, Rabia Altaf, Ashfaq Ahmad
JournalPharmaceutical biology (Pharm Biol) Vol. 52 Issue 11 Pg. 1411-22 (Nov 2014) ISSN: 1744-5116 [Electronic] England
PMID25026347 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Anti-Inflammatory Agents
  • Plant Extracts
Topics
  • Animals
  • Anti-Inflammatory Agents (isolation & purification, therapeutic use)
  • Azadirachta
  • Dose-Response Relationship, Drug
  • Drug Synergism
  • Edema (blood, drug therapy, pathology)
  • Female
  • Humans
  • Male
  • Plant Extracts (isolation & purification, therapeutic use)
  • Plant Leaves
  • Rats
  • Rats, Sprague-Dawley
  • Sheep
  • Treatment Outcome

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