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. 2022 Feb 11;11(2):364.
doi: 10.3390/antiox11020364.

Chemical Fingerprinting, Antioxidant, and Anti-Inflammatory Potential of Hydroethanolic Extract of Trigonella foenum-graecum

Hina Fatima  1   2   3 , Muhammad Shahid  1 , Chris Pruitt  2 , Meredith A Pung  2 , Paul J Mills  2 , Muhammad Riaz  4 , Rizwan Ashraf  5
Affiliations

Chemical Fingerprinting, Antioxidant, and Anti-Inflammatory Potential of Hydroethanolic Extract of Trigonella foenum-graecum

Hina Fatima et al. Antioxidants (Basel). .

Abstract

In the current study, the antioxidant and anti-inflammatory potential of hydroethanolic extract of T. foenum-graecum seeds was evaluated. Phenolic profiling of T. foenum-graecum was conducted through high-performance liquid chromatography-photodiode array (HPLC-PDA) as well as through the mass spectrometry technique to characterize compounds responsible for bioactivity, which confirmed almost 18 compounds, 13 of which were quantified through a chromatographic assay. In vitro antioxidant analysis of the extract exhibited substantial antioxidant activities with the lowest IC50 value of both DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) inhibition assays. The extract was found to be non-toxic against human RBCs and murine macrophage RAW 264.7 cells. Moreover, the extract significantly (p < 0.001) reduced the lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-α), intrlukin-6 (IL-6), prostaglandin E2 (PGE2), and nitric oxide (NO) in RAW 264.7 cells in a concentration-dependent manner. The hydroethanolic extract of T. foenum-graecum exhibited considerable anti-inflammatory potential by decreasing the cellular infiltration to the inflammatory site in both carrageenan-induced peritonitis and an air pouch model of inflammation. Pretreatment with T. foenum-graecum extract caused significant improvement in antioxidants such as superoxide dismutase (SOD), CAT (catalase), malondialdehyde (MDA), and myeloperoxidase (MPO) against oxidative stress induced by carrageenan. Based on our results of in vivo and in vitro experimentation, we concluded that hydroethanolic extract of T. foenum-graecum is a potential source of phenolic compounds with antioxidant and anti-inflammatory potential.

Keywords: T. foenum-graecum; air pouch inflammation; antioxidants; cellular infiltration; lipid peroxidation; oxidative stress markers; peritonitis.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
(A) Chromatogram of high-pressure liquid chromatography equipped with diode array detector (HPLC-DAD) at 280 nm and (B) extracted spectrum (200–400 nm) of each peak.
Figure 2
Figure 2
Purity plots of representative peaks, (A) gallic acid, and (B) trihydroxybenzoic acid.
Figure 3
Figure 3
UHPLC-Q-TOF chromatogram of T. foenum.
Figure 4
Figure 4
Hemolytic (a) and thrombolytic activity (b) of different doses (50–300 µg/mL) of hydroethanolic extract of T. foenum-graecum seeds through hemolytic and thrombolytic assays. (***, significant at p < 0.001).
Figure 5
Figure 5
Cytotoxic activity (cell viability) of hydroethanolic extract of T. foenum-graecum and doxorubicin (standard) against RAW 264.7 macrophage through MTT assay.
Figure 6
Figure 6
Anti-inflammatory profile of T. foenum-graecum seed extract through quantification of, TNF-α (A), IL-6 (B), NO (C) and PGE2 (D) measurement in RAW 264.7 cell’s culture medium after stimulation with LPS (1 µg/mL). +, treated with LPS, ***, significant at p < 0.001.
Figure 7
Figure 7
Anti-inflammatory profile (cellular infiltration) (A) (Total WBCs), (B) (Monocytes), (C) (Neutrophils) and Eosinophils (D) of hydroethanolic extract of T. foenum-graecum against air pouch model of inflammation. Data are expressed as mean ± S.D. ***, significant at p < 0.001.
Figure 8
Figure 8
Histopathological photographs of air pouch tissues after treatment with different concentrations of T. foenum-graecum extract at 100, 200, 400 mg/kg BW (A–C), carrageenan control (D), dexamethasone 20 mg/kg body weight (E) and normal control (F).
Figure 9
Figure 9
Anti-inflammatory effect A (Neutrophils) and B (Total WBCs) of hydroethanolic extract of T. foenum-graecum on carrageenan-induced peritonitis. Rats were pretreated orally with different concentrations of T. foenum-graecum extract and dexamethasone (20 mg/kg body weight). Data are expressed as mean ± SD. ***, significant at p < 0.001.
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