Nigella sativa oil protects against cadmium-induced intestinal toxicity via promotion of anti-inflammatory mechanisms, mucin expression and microbiota integrity

Document Type : Original Research Article

Authors

1 Environmental and Gastrointestinal Toxicology Laboratory, Department of Veterinary Physiology and Biochemistry, Faculty of Veterinary Medicine, University of Ibadan, Oyo State, Nigeria

2 Environmental Microbiology and Biotechnology Laboratory, Department of Microbiology, Faculty of Science, University of Ibadan, Oyo State, Nigeria

3 Department of Chemistry, Faculty of Science, The Polytechnic, Ibadan, Oyo State, Nigeria

Abstract

Objective: This study examined the protective effects of Nigella sativa oil (NSO) on cadmium (Cd)-induced alterations affecting gut morphology and microbiota composition, as well as the involvement of mucus glycoprotein (MUC2) and immuno-inflammatory markers (TNFα and IL-2) in the colon of rats.
Materials and Methods: Male Wistar rats, randomized into four groups, were treated either with distilled water (control), CdCl2 (100 mg/kg), CdCl2+NSO (1 ml/kg) or NSO alone. After the experiments, faecal samples were processed for microbial culture on various selective media, while intestinal segments were prepared for histopathological examination and immunohistochemistry. The composition of NSO was analyzed using Gas Chromatography-Mass Spectrometry (GC-MS).
Results: Oral Cd administration provoked dramatic increases in faecal counts of potentially pathogenic bacteria (Staphylococci, Enterococci, Pseudomonas and Escherichia coli), while decreasing probiotic lactobacilli counts. Cadmium treatment caused down-regulation of colonic MUC2 (p=0.003) and IL-2 (p=0.03), but increased TNFα (p=0.034), along with reduced goblet cell counts and mucus production. Conversely, treatment with NSO significantly improved Lactobacilli counts (p=0.042), while reducing the levels of potentially pathogenic species. In addition, NSO significantly restored colonic expressions of MUC2 (p=0.001), TNFα (p=0.007) and IL-2 (p=0.025) to control levels. GC-MS analysis of NSO revealed the presence of the active ingredient, thymoquinone and a high content of unsaturated fatty acids, including trans-13-octadecenoic acid and oleic acid.
Conclusion: This study highlights the intestinal mucus, microbiota and immuno-inflammatory system as important protective targets of NSO against Cd-induced intestinal toxicity.

Keywords


Ahmad A, Husain A, Mujeeb M, Khan SA,
Najmi AK, Siddique NA, Damanhouri ZA,
Anwar F. 2013. A review on therapeutic
potential of Nigella sativa: A miracle herb.
Asian Pac J Trop Med, 3: 337-352.
Akinrinde AS, Adebiyi OE, Asekun A. 2019.
Amelioration of Aflatoxin B1-induced
gastrointestinal injuries by Eucalyptus oil
in rats. J Complement Integr Med 17.
Alenzi FQ, El-Bolkiny YE, Salem ML. 2010.
Protective effects of Nigella sativa oil and
thymoquinone against toxicity induced by
the anticancer drug cyclophosphamide. Br J
Biomed Sci, 67: 20-28.
Alzoghaibi M. 2007. Protective effect of oleic
acid against acute gastric mucosal lesions
induced by ischemia-reperfusion in rat.
Saudi J Gastroenterol, 13: 17-20.
Asagba SO. 2013. Cadmium Absorption. In:
Kretsinger R.H., Uversky V.N., Permyakov
E.A. (eds) Encyclopedia of
Metalloproteins. Springer, New York, NY.
Avwioro OG. 2010. Histochemistry and
Tissue pathology, Principle and
Techniques, Claverianum press, Nigeria.
Cameron EA, Sperandio V. 2015. Frenemies:
signaling and nutritional integration in
pathogen-microbiota-host interactions. Cell
Host Microbe, 18: 275-284.
Candido FG, Valente FX, Grzeskowiak LM,
Moreira APB, Rocha DMUP, Alfenas RG.
2017. Impact of dietary fat on gut
microbiota and low-grade systemic
inflammation: mechanisms and clinical
implications on obesity. Int J Food Sci
Nutr, 69: 125-143.
Chaieb K, Kouidhi B, Jrah H, Mahdouani K,
Bakhrouf A. 2011. Antibacterial activity of
Thymoquinone, an active principle of
Nigella sativa and its potency to prevent
bacterial biofilm formation. BMC
Complement Altern Med, 13: 11-29.
Chauhan SS, Ojha S, Mahmood A. 2011.
Modulation of lipid peroxidation and
antioxidant defense systems in rat intestine
by sub-chronic fluoride and ethanol
administration. Alcohol, 45: 663-672.
Dajani EZ, Shahwan TG, Dajani NE. 2016.
Overview of the preclinical
pharmacological properties of Nigella
sativa (black seeds): a complementary drug
with historical and clinical significance. J
Physiol Pharmacol, 67: 801-817.
Donaldson GP, Ladinsky MS, Yu KB, Sanders
JG, Yoo B, Chou W-C, Conner ME, Earl
Nigella sativa oil mitigates cadmium-induced intestinal toxicity
AJP, Vol. 12, No. 3, May-Jun 2022 255
AM, Knight R, Bjorkman PJ, Mazmanian
SK. 2018. Gut microbiota utilize
immunoglobulin A for mucosal
colonization. Science, 360: 795-800.
Drury RA, Wallington EA. 1976. Carlton‟s
Histopathological Techniques, 4th ed.
Oxford University Press, London, pp. 139-
142.
Elhady K, Abdelhamid A, Elgawish R. 2012.
Effect of cadmium chloride on interleukin2, lymphocyte and DNA in adult male
albino rats. Toxicol Lett, 211: S140-S140.
Gillois K, Leveque M, Theodorou V, Robert
H, Mercier-Bonin M. 2018. Mucus: An
underestimated gut target for environmental
pollutants and food additives.
Microorganisms, 6: 53.
Grondin JA, Kwon YH, Far PM, Haq S, Khan
WI. 2020. Mucins in intestinal mucosal
defense and inflammation: Learning from
clinical and experimental studies. Front
Immunol, 11: 2054.
Harrigan WF, MacCance ME. 1996.
Laboratory Methods in Microbiology,
Academic Press.
Hsu H-P, Lai M-D, Lee J-C, Yen M-C, Weng
T-Y, Chen W-C, Fang J-H, Chen Y-L.
2017. Mucin 2 silencing promotes colon
cancer metastasis through interleukin-6
signaling. Sci Rep, 7: 5823.
Isik S, Erdem SA, Kartal M. 2019.
Investigation of the fatty acid profile of
commercial black cumin seed oils and seed
oil capsules: Application to real samples. J
Chem Metrol, 13: 53-60.
Jiang WW, Wang QH, Liao YJ, Peng P, Xu
M, Yin L-X. 2017. Effects of
dexmedetomidine on TNF-α and
interleukin-2 in serum of rats with severe
cranio-cerebral injury. BMC Anesthesiol,
17: 130.
Jin Y, Wu S, Zeng Z, Fu Z. 2017. Effects of
environmental pollutants on gut microbiota.
Environ Pollut, 222: 1-9.
Khan MA, Afzal M. 2016. Chemical
composition of Nigella sativa Linn: Part 2
Recent Advances. Inflammaopharmacol,
24: 67-79.
Kim JJ, Khan WI. 2013. Goblet cells and
Mucins: Role in innate defense in enteric
infections. Pathogens, 2: 55-70.
Larsson JMH, Karlsson H, Crespo
JG, Johansson MEV, Eklund L, Sjövall
H, Hansson GC. 2011. Altered Oglycosylation profile of MUC2 mucin
occurs in active ulcerative colitis and is
associated with increased inflammation.
Inflamm. Bowel Dis, 17: 2299-2307.
Li H, Limenitakis JP, Fuhrer T, Geuking MB,
Lawson MA, Wyss M, Brugiroux S, Keller
I, Macpherson JA, Rupp S, Stolp B, Stein J,
Stecher B, Sauer U, McCoy K. 2015. The
outer mucus layer acts hosts a distinct
intestinal microbial niche. Nat Comm, 6:
8292.
Liu Y, Li Y, Liu K, Shen J. 2014. Exposing to
cadmium stress cause profound toxic effect
on microbiota of the mice intestinal tract.
PLoS One, 9: e85323.
Mosca A, Leclerc M, Hugot JP. 2016. Gut
microbiota diversity and human diseases:
Should we reintroduce key predators in our
ecosystem. Front Microbiol, 7: 455.
Nagpal R, Yadav H. 2017. Bacterial
translocation from the gut to the distant
organs: An overview. Ann Nutr Metab, 71:
11-16.
Osifo CU, Iyawe VI. 2018. Cadmium
exposure induces early catabolism in male
Wistar rat experiment. J Mol Cell Biochem,
2: 9.
Paone P, Cani PD. 2020. Mucus barrier,
mucins and gut microbiota: the expected
slimy partners? Gut, 69: 2232-2243.
Parameswaran N, Patial S. 2010. Tumor
necrosis factor-α signaling in macrophages.
Crit Rev Eukaryot Gene Expr, 20: 87-103.
Perry RJ, Resch JM, Douglass AM, Madara
JC, Rabin-Court A, Kucukdereli H, Wu C,
Song JD, Lowell BB, Shulman GI. 2019.
Leptin‟s hunger-suppressing effects are
mediated by the hypothalamic-pituitaryadrenocortical axis in rodents. Proc Natl
Acad Sci USA, 116: 13670-13679.
Phillipson M, Atuma C, Henriksnäs J, Holm L.
2002. The importance of mucus layers and
bicarbonate transport in preservation of
gastric juxta-mucosal pH. Am J Physiol
Gastrointest Liver Physiol, 282: G211-
G2G9.
Public Health Service (PHS). 1996. Public
health service policy on humane care and
the use of laboratory animals. Washington,
DC: US Department of Health and Humane
Services, Pp 99-158.
Roberfroid M, Gibson GR, Hoyles L,
McCartney AL, Rastall R, Rowland I,
Wolvers D, Watzl B, Szajewska H, Stahl B,
Guarner F, Respondek F, Whelan K,
Coxam V, Davicco M-J, Léotoing L,
Akinrinde et al.
AJP, Vol. 12, No. 3, May-Jun 2022 256
Wittrant Y, Delzenne N, Cani P, Neyrinck
A, Meheust A. 2010. Prebiotic effects,
metabolic and health benefits. Br J Nutr,
12: 1-16.
Schreiber O, Petersson J, Walde´n T, Ahl D,
Sandler S, Phillipson M, Holm L. 2013.
iNOS-dependent Increase in Colonic
Mucus Thickness in DSS-Colitic Rats.
PLoS ONE, 8: e71843.
Sicard J-F, Le Bihan G, Vogeleer P, Jacques
M, Harel J. 2017. Interactions of intestinal
bacteria with components of the intestinal
mucus. Front Cell Infect Microbiol, 7: 387.
Tinkov AA, Gritsenko VA, Skalnaya MG,
Cherkasov SV, Aaseth J, Skalny AV. 2018.
Gut as a target for cadmium toxicity.
Environ Pollut, 235: 429-434.
Todorich B, Olopade JO, Surguladze N, Zhang
X, Neely E, Connor JR. 2011. The
mechanism of vanadium-mediated
developmental hypomyelination is related
to destruction of oligodendrocyte
progenitors through a relationship with
ferritin and iron, Neurotoxic Res, 19: 361-
373.
van Vilet MJ, Harmsen HJ, de Bont ES,
Tissing WJ. 2010. The role of intestinal
microbiota in the development and severity
of chemotherapy-induced mucositis. PLoS
Pathog, 6: e1000879.
Varum FJO, Veiga F, Sousa JS, Basit AW.
2011. Mucus thickness in the
gastrointestinal tract of laboratory animals.
J Pharm Pharmacol, 64: 218-227.
Venegas DP, De la Fuente MK, Landskron G,
Gonzalez MJ, Quera R, Dijkstra G,
Harmsen HJM, Faber KN, Hermoso MA.
2019. Short chain fatty acids (SCFAs)-
mediated gut epithelial and immune
regulation and its relevance for
inflammatory bowel diseases. Front
Immunol, 10: 277.
Wang H, Hou L, Kwak D, Fassett J, Xu X,
Chen A, Chen W, Blazar BR, Xu Y, Hall
JL, Ge JB, Bache RJ, Chen Y. 2016.
Increasing regulatory t cells with
interleukin-2 and interleukin-2 antibody
attenuates lung inflammation and heart
failure progress. Hypertension, 68: 114-
122.