Trigonella foenum-graecum Seeds Oil Attenuated Inflammation and Angiogenesis in vivo through Down-Regulation of TNF-α
Authors: Asif Muhammad , Yousaf Hafiz M. , Saleem Mohammad , Saadullah Malik , Chohan Tahir A. , Shamas Muhammad U. , Yaseen Hafiza S. , Mahrukh  , Yousaf Muhammad U. , Yaseen Maria 
Published: October 2020

Introduction:
Inflammation is a vital reaction of the natural immune system that protects against
encroaching agents. However, uncontrolled inflammation can lead to complications. Trigonella foenumgraecum
is traditionally used as an anti-inflammatory herb.

Objectives:
The current study was conducted to explore the antioxidant, anti-inflammatory, and antiangiogenic
potentials of Trigonella foenum-graecum seeds oil.

Methods:
Oil was extracted from seeds of Trigonella foenum-graecum by cold press method and labelled as
TgSO. Phytochemical (GC-MS, Folin-Ciocalteu method) and metal analyses were conducted to evaluate the
metalo-chemical profile of TgSO. In vitro antioxidant assays (2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis-3-
ethylbenzothiazoline-6-sulfonic acid and ferric reducing antioxidant power) were performed to assess its antioxidant
potential. In vitro antimicrobial activity was evaluated using agar disc diffusion method and the safety
profile of TgSO was assessed in acute toxicological studies following OECD 425 guidelines. In vivo antiinflammatory
activities of TgSO were assessed in carrageenan, serotonin, histamine, formalin, and cotton pelletinduced
oedema models. Serum TNF-α, Superoxide Dismutase (SOD) and, Catalases (CAT) levels were assessed
by ELISA kits. In vivo antiangiogenic activity of TgSO was screened in chick Chorioallantoic Membrane
(CAM) assay. Histopathological studies using excised paws were conducted to observe the effects of TgSO
treatment at the tissue level. In silico docking studies were conducted to screen the binding potentials of identified
compounds with TNF-α.

Results:
Extraction by cold press method yielded 16% of TgSO. Phytochemical analysis of TgSO through GCMS
showed the presence of eugenol, dihydrocoumairn, heptadecanoic acid, tri- and tetradecanoic acid, and
hexadecanoic acid, respectively. Total phenolic contents of TgSO were found to be 0.30±0.01mg/g gallic acid
equivalent in Folin-Ciocalteu method. Metal analysis indicated the presence of different metals in TgSO. Findings
of antioxidant models showed the moderate antioxidant potential of TgSO. Findings of antimicrobial assays
showed that TgSO was active against bacterial (S. aureus, S. epidermidis) and fungal (C. albicans, and A. niger)
strains. In vivo toxicity study data showed that TgSO was safe up to the dose of 5000 mg/kg. Data of oedema
models showed a significant (p<0.05) reduction in oedema development in TgSO treated animals in both acute
and chronic models. Histopathological evaluations of paws showed minimum tissue infiltration with inflammatory
cells in TgSO-treated animals. Treatment with TgSO also significantly (p<0.05) down-regulated TNF-α in
serum while levels of SOD and CAT were up-regulated. Findings of the CAM assay revealed the antiangiogenic
activity of TgSO. Findings of in silico docking studies showed that identified phytoconstituents can bind with
culprit cytokine (TNF-α).

Conclusion:
Data obtained from the current study conclude that TgSO has antioxidant, anti-inflammatory, and
antiangiogenic effects that validate its traditional uses. Synergistic actions of different phytoconstituents are
proposed to be responsible for the observed effects.