Skip to main content
SpringerLink
Log in

Investigation of bioactive fractions from fenugreek seeds (Trigonella foenum-graecum L.): chemical profiling, in vitro anti-inflammatory activity, and their potential against LPS-stimulated J774A.1 cells

  • Research Article
  • Published:
Advances in Traditional Medicine Aims and scope Submit manuscript

Abstract

Despite the known anti-inflammatory effect of fenugreek seeds (Trigonella foenum-graecum L.), the specific mechanism and compounds responsible for this effect have been less studied so far. This research aimed to fractionate the total methanolic extract and investigate their impact on the iNOS expression in J774A.1 macrophage induced by lipopolysaccharide (LPS). The fractions of the total extract were obtained using semi-preparative HPLC. J774A.1 cells were treated with the highest non-toxic concentration of the samples and bacterial LPS (5 μg/ml). After 24 h, treated and untreated cells were used for RNA extraction, and mRNA levels were determined by qRT-PCR analysis. A total of 30 fractions were obtained with 9 main fractions (F3, F6, F9, F12, F15, F18, F21, F24, and F27) selected for cell studies. F9, total extract, and F15 significantly decreased iNOS gene expression compared to the LPS-treated group (up to 99.3%, 84.7% and 76.5% respectively). Overall, these results suggest that the anti-inflammatory effect of fenugreek seeds and/or fractions may be partially due to the suppression of iNOS. This effect can be attributed to various plant compounds, and further isolation may help identify novel anti-inflammatory compounds become a new inflammation. These findings support the potential of both the extract and the fractions as promising candidates for the treatment of inflammation-related diseases in future research.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Data availability

The data that support the findings of this study are available in the manuscript.

References

  • Camacho-Barquero L, Villegas I, Sánchez-Calvo JM, Talero E, Sánchez-Fidalgo S, Motilva V et al (2007) Curcumin, a Curcuma longa constituent, acts on MAPK p38 pathway modulating COX-2 and iNOS expression in chronic experimental colitis. Int Immunopharmacol 7(3):333–342

    Article  CAS  PubMed  Google Scholar 

  • Chan MM-Y, Huang H-I, Fenton MR, Fong D (1998) In vivo inhibition of nitric oxide synthase gene expression by curcumin, a cancer preventive natural product with anti-inflammatory properties. Biochem Pharmacol 55(12):1955–1962

    Article  CAS  PubMed  Google Scholar 

  • Chen L, Yang S, Zumbrun EE, Guan H, Nagarkatti PS, Nagarkatti M (2015) Resveratrol attenuates lipopolysaccharide-induced acute kidney injury by suppressing inflammation driven by macrophages. Mol Nutr Food Res 59(5):853–864

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Choi K-C, Son Y-O, Hwang J-M, Kim B-T, Chae M, Lee J-C (2017) Antioxidant, anti-inflammatory and anti-septic potential of phenolic acids and flavonoid fractions isolated from Lolium multiflorum. Pharm Biol 55(1):611–619

    Article  CAS  PubMed  Google Scholar 

  • Choy KW, Murugan D, Leong X-F, Abas R, Alias A, Mustafa MR (2019) Flavonoids as natural anti-inflammatory agents targeting nuclear factor-kappa B (NFκB) signaling in cardiovascular diseases: a mini review. Front Pharmacol 10:1295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • García-Mediavilla V, Crespo I, Collado PS, Esteller A, Sánchez-Campos S, Tuñón MJ et al (2007) The anti-inflammatory flavones quercetin and kaempferol cause inhibition of inducible nitric oxide synthase, cyclooxygenase-2 and reactive C-protein, and down-regulation of the nuclear factor kappaB pathway in Chang Liver cells. Eur J Pharmacol 557(2–3):221–229

    Article  PubMed  Google Scholar 

  • González R, Ballester I, López-Posadas R, Suárez M, Zarzuelo A, Martínez-Augustin O et al (2011) Effects of flavonoids and other polyphenols on inflammation. Critic Rev Food Sci Nutr 51(4):331–362

    Article  Google Scholar 

  • Harris Daniel C (1988) Quantitative Chem Analysis. 8th edition. W.H. Freeman Pub

  • Hamalainen M, Nieminen R, Vuorela P, Heinonen M, Moilanen E (2007) Anti-inflammatory effects of flavonoids: genistein, kaempferol, quercetin, and daidzein inhibit STAT-1 and NF-κB activations, whereas flavone, isorhamnetin, naringenin, and pelargonidin inhibit only NF-κB activation along with their inhibitory effect on iNOS expression and NO production in activated macrophages. Med Inflamm 2007:045673

  • Kamal P, Pal R, Nath R, Sachan AK (2022) Protective effects of the ethanolic fenugreek seeds extract and its potentiation with nitric oxide modulators in adjuvant induced changes in arthritic index, proinflammatory/anti-inflammatory cytokines imbalance and oxidative stress markers in rats. Indian J Physiol Pharmacol Ther 66(1):45–54

    Article  CAS  Google Scholar 

  • Kassaian N, Azadbakht L, Forghani B, Amini M (2009) Effect of fenugreek seeds on blood glucose and lipid profiles in type 2 diabetic patients. Int J Vitam Nutr Res 79(1):34–39

    Article  CAS  PubMed  Google Scholar 

  • Kawabata T, Cui M-Y, Hasegawa T, Takano F, Ohta T (2011) Anti-inflammatory and anti-melanogenic steroidal saponin glycosides from Fenugreek (Trigonella foenum-graecum L.) seeds. Planta Med 77(07):705–710

    Article  CAS  PubMed  Google Scholar 

  • Kim HP, Son KH, Chang HW, Kang SS (2004) Anti-inflammatory plant flavonoids and cellular action mechanisms. J Pharmacol Sci 96(3):229–245

    Article  CAS  PubMed  Google Scholar 

  • Kim J-Y, Park SJ, Yun K-J, Cho Y-W, Park H-J, Lee K-T (2008) Isoliquiritigenin isolated from the roots of Glycyrrhiza uralensis inhibits LPS-induced iNOS and COX-2 expression via the attenuation of NF-κB in RAW 264.7 macrophages. Eur J Pharmacol 584(1):175–184

    Article  CAS  PubMed  Google Scholar 

  • Lin C-Y, Wang W-H, Chen S-H, Chang Y-W, Hung L-C, Chen C-Y et al (2017) Lipopolysaccharide-induced nitric oxide, prostaglandin E2, and cytokine production of mouse and human macrophages are suppressed by pheophytin-b. Int J Mol Sci 18(12):2637

    Article  PubMed  PubMed Central  Google Scholar 

  • Liu Y, Kakani R, Nair MG (2012) Compounds in functional food fenugreek spice exhibit anti-inflammatory and antioxidant activities. Food Chem 131(4):1187–1192

    Article  CAS  Google Scholar 

  • Mandegary A, Pournamdari M, Sharififar F, Pournourmohammadi S, Fardiar R, Shooli S (2012) Alkaloid and flavonoid rich fractions of fenugreek seeds (Trigonella foenum-graecum L.) with antinociceptive and anti-inflammatory effects. Food Chem Toxicol 50(7):2503–2507

    Article  CAS  PubMed  Google Scholar 

  • Mehrabani M, Raeiszadeh M, Najafipour H, Esmaeli Tarzi M, Amirkhosravi A, Poustforoosh A et al (2020) Evaluation of the cytotoxicity, antibacterial, antioxidant, and anti-inflammatory effects of different extracts of Punica granatum var. pleniflora. JKMU 27(5):414–425

    Google Scholar 

  • Miri A, Sarani M, Najafidoust A, Mehrabani M, Zadeh FA, Varma RS (2022) Photocatalytic performance and cytotoxic activity of green-synthesized cobalt ferrite nanoparticles. Mater Res Bull 149:111706

    Article  CAS  Google Scholar 

  • Mohamadi N, Pournamdari M, Sharififar F, Ansari M (2020) Simultaneous spectrophotometric determination of trigonelline, diosgenin and nicotinic acid in dosage forms prepared from fenugreek seed extract. IJPR 19(2):153

    CAS  PubMed  PubMed Central  Google Scholar 

  • Mohamadi N, Sharififar F, Ansari M, Pournamdari M, Rezaei M, Hassanabadi N (2021) Pharmacokinetic profile of diosgenin and trigonelline following intravenous and oral administration of fenugreek seed extract and pure compound in rabbit. J Asian Nat Prod Res 23(5):466–477

    Article  CAS  PubMed  Google Scholar 

  • Morgan MJ, Liu Z-G, Cells (2010) Reactive oxygen species in TNFα-induced signaling and cell death. Mol Cells 30(1):1–12

    Article  CAS  PubMed  Google Scholar 

  • Naidu MM, Shyamala B, Naik JP, Sulochanamma G, Srinivas P (2011) Chemical composition and antioxidant activity of the husk and endosperm of fenugreek seeds. LWT - Food Sci Technol 44(2):451–456

    Article  Google Scholar 

  • Noudeh GD, Sharififar F, Noodeh AD, Moshafi MH, Afzadi MA, Behravan E et al (2010) Antitumor and antibacterial activity of four fractions from Heracleum persicum Desf. and Cinnamomum zeylanicum Blume. J Med Plants Res 4(21):2176–80

    Google Scholar 

  • Pandian RS, Anuradha C, Viswanathan P (2002) Gastroprotective effect of fenugreek seeds (Trigonella foenum graecum) on experimental gastric ulcer in rats. J Ethnopharmacol 81(3):393–397

    Article  PubMed  Google Scholar 

  • Predonzani A, Calì B, Agnellini AH, Molon B (2015) Spotlights on immunological effects of reactive nitrogen species: when inflammation says nitric oxide. WJEM, World J Exp Med 5(2):64. https://doi.org/10.5493/wjem.v5.i2.64

    Article  PubMed  Google Scholar 

  • Pundarikakshudu K, Shah DH, Panchal AH, Bhavsar GC (2016) Anti-inflammatory activity of fenugreek (Trigonella foenum-graecum Linn) seed petroleum ether extract. Indian J Pharmacol 48(4):441–444

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Raeiszadeh M, Esmaeili-Tarzi M, Bahrampour-Juybari K, Nematollahi-Mahani S, Pardakhty A, Nematollahi M et al (2018) Evaluation the effect of Myrtus communis L. extract on several underlying mechanisms involved in wound healing: an in vitro study. S Afr J Bot 118:144–150

    Article  Google Scholar 

  • Samareh Fekri M, Mandegary A, Sharififar F, Poursalehi HR, Nematollahi MH, Izadi A et al (2018) Protective effect of standardized extract of Myrtus communis L.(myrtle) on experimentally bleomycin-induced pulmonary fibrosis: biochemical and histopathological study. Drug Chem Toxicol 41(4):408–414

    Article  CAS  PubMed  Google Scholar 

  • Sarhadynejad Z, Sharififar F, Pardakhty A, Nematollahi M-H, Sattaie-Mokhtari S, Mandegary A (2016) Pharmacological safety evaluation of a traditional herbal medicine “Zereshk-e-Saghir” and assessment of its hepatoprotective effects on carbon tetrachloride induced hepatic damage in rats. J Ethnopharmacol 190:387–395

    Article  PubMed  Google Scholar 

  • Sharififar F, Khazaeli P, Alli N (2009) In vivo evaluation of anti-inflammatory activity of topical preparations from Fenugreek (Trigonella foenum-graecum L.) seeds in a cream base. Iran J Pharm Sci 5(3):157–162

    Google Scholar 

  • Sheu F, Lai H-H, Yen G-C (2001) Suppression effect of soy isoflavones on nitric oxide production in RAW 264.7 macrophages. J Agric Food Chem 49(4):1767–1772

    Article  CAS  PubMed  Google Scholar 

  • Sindhu G, Ratheesh M, Shyni G, Nambisan B, Helen A (2012) Anti-inflammatory and antioxidative effects of mucilage of Trigonella foenum graecum (Fenugreek) on adjuvant induced arthritic rats. Int Immunopharmacol 12(1):205–211

    Article  CAS  PubMed  Google Scholar 

  • Tavakoly R, Maracy MR, Karimifar M, Entezari MH (2018) Does fenugreek (Trigonella foenum-graecum) seed improve inflammation, and oxidative stress in patients with type 2 diabetes mellitus? A parallel group randomized clinical trial. Eur J Integr Med 18:13–17

    Article  Google Scholar 

  • Torre D, Pugliese A, Speranza F (2002) Role of nitric oxide in HIV-1 infection: friend or foe? Lancet Infect Dis 2(5):273–280

    Article  CAS  PubMed  Google Scholar 

  • Tripathi P, Tripathi P, Kashyap L, Singh V (2007) The role of nitric oxide in inflammatory reactions. FEMS Immunol Med Microbiol Immunol 51(3):443–452

    Article  CAS  Google Scholar 

  • Tsai SH, Lin-Shiau SY, Lin JK (1999) Suppression of nitric oxide synthase and the down-regulation of the activation of NFκB in macrophages by resveratrol. Br J Pharmacol Chemother 126(3):673–680

    Article  CAS  Google Scholar 

  • Vasanthkumar T, Hanumanthappa M, Lakshminarayana R (2019) Curcumin and capsaicin modulates LPS induced expression of COX-2, IL-6 and TGF-β in human peripheral blood mononuclear cells. Cytotechnology 71:963–976

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zamora R, Vodovotz Y, Billiar TR (2000) Inducible nitric oxide synthase and inflammatory diseases. Mol Med 6(5):347–373

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

I am thankful for the resources provided by Vice Chancellor for Research and Technology, Kerman University of Medical Science that enabled the successful execution of this project.

Funding

This study was funded by Kerman University of Medical Science, Kerman, Iran (Grant number: 95000275.

Author information

Authors and Affiliations

  1. Department of Clinical Pharmacy, Shiraz School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran

    Ensieh Kalantari

  2. Department of Medicinal Chemistry, Pharmaceutics Research Center, Neuropharmacology Institute, Kerman University of Medical Sciences, Kerman, Iran

    Mostafa Pournamdari

  3. Physiology Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran

    Mehrnaz Mehrabani & Mojdeh Esmaeili Tarzic

  4. Herbal and Traditional Medicines Research Center, Kerman University of Medical Sciences, Kerman, Iran

    Neda Mohamadi, Amir Asadi, Sheida Illaghi & Fariba Sharififar

Authors
  1. Ensieh Kalantari
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mostafa Pournamdari
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mehrnaz Mehrabani
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mojdeh Esmaeili Tarzic
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Neda Mohamadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Amir Asadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Sheida Illaghi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Fariba Sharififar
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Mostafa Pournamdari or Fariba Sharififar.

Ethics declarations

Ethical statement

This article does not contain any studies with human participants or animals performed by any of the authors.

Conflict of interest

Ensieh Kalantari has no conflict of interest. Mostafa Pournamdari has no conflict of interest. Mehrnaz Mehrabani has no conflict of interest. Mojdeh Esmaeili Tarzic has no conflict of interest. Neda Mohamadi has no conflict of interest. Amir Asadi has no conflict of interest. Sheida Illaghi has no conflict of interest. Fariba Sharififar has no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Kalantari, E., Pournamdari, M., Mehrabani, M. et al. Investigation of bioactive fractions from fenugreek seeds (Trigonella foenum-graecum L.): chemical profiling, in vitro anti-inflammatory activity, and their potential against LPS-stimulated J774A.1 cells. ADV TRADIT MED (ADTM) (2024). https://doi.org/10.1007/s13596-024-00751-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s13596-024-00751-4

Keywords

Search

Navigation