Investigation of the Effect of Sulforaphane Content of Brassica oleracea Extract on the Expression of NF-κB and STAT3 Genes Contributing Affecting the Inflammatory Response Pathway of Immune System

Document Type : Research Paper

Authors

1 M.Sc., Department of Agricultural Biotechnology, Payame Noor University, Tehran, Iran.

2 Assistant Professor, Department of Agricultural Biotechnology, Payame Noor University, Tehran, Iran.

Abstract

Inflammation is a natural response of the body’s immune system to disease, protecting the body in the form of a defensive mechanism. The immune system identifies the damaged body cells, stimuli, and pathogens to trigger the healing process – a mechanism that can engage the body with inflammation. In some cases, too long inflammation and excessive immune responses led to side effects and damages that dominate their benefits, leading to chronic inflammation and then engagement with different diseases. In the present research, effect of the broccoli sprouts extract on the expression of the NF-κB and STAT3 genes affecting the inflammatory response pathway of immune system was investigated. Results of gas chromatography – mass spectroscopy (GC-MS) and MTT assay confirmed the presence of the sulforaphane in the extract while rejecting their toxicity to cell lines. Moreover, the GAPDH and β-actin genes were used as internal control genes in the 1321N1 neural cell line, with the HFF cell line used as normal cell. The cells were treated with 2 mg of the extract for 3 days, during which time the changes in the immune system in terms of the genes expressions were monitored. The results showed that the sulforaphane attenuate the expressions of the NF-κB and STAT3 genes as two of the most important inflammatory mediating genes. It seems that the use of sulforaphane as active ingredient can effectively attenuate the cause of the disorders faced by the patients with immune system inflammation.

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Auditeau E, Chassagne F, Bourdy G, Bounlu M, Jost J, Luna J, Boumediene F (2019) Herbal medicine for epilepsy seizures in Asia, Africa and Latin America: A systematic review Journal of ethnopharmacology 234: 119-153. Balkwill F, Charles KA, Mantovani A (2005) Smoldering and polarized inflammation in the initiation and promotion of malignant disease.211–217. Cai Q, Lin J, Wei L, Zhang L, Wang L, Zhan Y, Peng J, (2012) Hedyotis diffusa Willd inhibits colorectal cancer growth in vivo via inhibition of STAT3 signaling pathway. International Journal of Molecular Sciences 13(5): 6117-6128. Coussens LM, Werb Z (2002) Inflammation and cancer. Nature. 860–867. Chiu CT, Chen JH, Chou FP, Lin HH (2015) Hibiscus sabdariffa leaf extract inhibits human prostate cancer cell invasion via down-regulation of Akt/NF-kB/MMP-9 pathway. Nutrients 7(7): 5065-5087. Fahey J, W Zhang Y, Talalay P (1997) Broccoli sprouts: an exceptionally rich source of inducers of enzymes that protect against chemical 005 carcinogens. Proceedings of the National Academy of Sciences. 94(19): 10367-10372. Fathy SA, Emam MA, Agwa SA, Zahra FA, Youssef FS, Sami RM (2016) The antiproliferative effect of Origanum majorana on human hepatocarcinoma cell line: suppression of NF-kB. Cellular and Molecular Biology. 62(10): 80-84. Fiandalo MV, Kyprianou N (2012) Caspase control: protagonists of cancer cell apoptosis. Experimental Oncology. 34(3): 165. Garbett K, Ebert PJ, Mitchell A, Lintas C, Manzi B, Mirnics K, Persico AM (2008) Immune transcriptome alterations in the temporal cortex of subjects with autism. Neurobiology of Disease. 30(3): 303-311. Gilliland G, Perrin S, Bunn HF (1990) Competitive PCR for quantitation of mRNA. PCR protocols: A guide to methods and applications. 60-69. Giovannoni SJ, Britschgi TB, Moyer CL, Field KG (1990) Genetic diversity in Sargasso Sea bacterioplankton. Nature. 345(6270): 60-63. Goines PE, Croen LA, Braunschweig D, Yoshida CK, Grether J, Hansen R, Kharrazi M, Ashwood P, Van de Water J (2011) Increased midgestational IFN-c, IL-4 and IL-5 in women bearing a child with autism: a case–control study. Mol. Autism.2:13. Hahm ER, Singh SV(2010) Sulforaphane inhibits constitutive and Interleukin-6–induced activation of Signal Transducer and Activator of Transcription 3 in prostate cancer cells. Cancer Prevention Research. 3(4): 484-494. Higdon JV, Delage B, Williams DE, Dashwood RH (2007) Cruciferous vegetables and human cancer risk epidemiologic evidence and mechanistic basis. Pharmacol Res. 55(3): 224-236 Hsuan CF, Hsu HF, Tseng WK, Lee TL, Wei YF, Hsu KL, Houng JY (2015) Glossogyne tenuifolia extract inhibits TNF-α-induced expression of adhesion molecules in human umbilical vein endothelial cells via blocking the NF-kB signaling pathway. Molecules 20(9): 16908-16923. Kane A, Deenick EK, Ma CS, Cook MC, Uzel G, Tangye SG (2014) STAT3 is a central regulator oflymphocyte differentiation and function. Curr. Opin. Immunol. 28:49-57. Kumar A, Takada Y, Boriek AM, Aggarwal BB (2004) Nuclear factor-kappaB: its role in health and disease. J. Mol. Med. 434–448 Kim SR, Seo HS, Choi HS, Cho SG, Kim YK, Hong EH, Ko SG, (2013) Trichosanthes kirilowii ethanol extract and cucurbitacin D inhibit cell growth and induce apoptosis through inhibition of STAT3 activity in breast cancer cells. Evidence-Based Complementary and Alternative Medicine, 2013. Lin M, Lin J, Wei L, Xu W, Hong Z, Cai Q, Zhu D (2012) Hedyotis diffusa Willd extract inhibits HT 29 cell proliferation via cell cycle arrest. Experimental and Therapeutic Medicine. 4(2): 307-310. Lin J, Li Q, Chen H, Lin H, Lai Z, Peng J, (2015) Hedyotis diffusa Willd. extract suppresses proliferation and induces apoptosis via IL-6-inducible STAT3 pathway inactivation in human colorectal cancer cells. Oncology Letters. 9(4): 1962-1970. Liu W, Ge T, Pan Z, Leng Y, Lv J, Li B (2017) The effects of herbal medicine on epilepsy. Oncotarget. 8(29): 48385. Mallik BK, Panda T, Padhy RN (2012) Traditional herbal practices by the ethnic people of Kalahandi district of Odisha, India. Asian Pac. J. Trop. Biomed. 2, S988–S994. Mohamed AAR, Metwally MM, Khalil SR, Salem GA, Ali HA (2019) Moringa oleifera extract attenuates the CoCl2 induced hypoxia of rat's brain: expression pattern of HIF-1α, NF-kB, MAO and EPO. Biomedicine & Pharmacotherapy. 109: 1688-1697. Nathan C (2002) Points of control in inflammation. Nature. 846–852. Newcombe EA, Camats-Perna J, Silva ML, Valmas N, Huat TJ, Medeiros R. (2018) Inflammation: the link between comorbidities, genetics, and Alzheimer’s disease. Journal of neuroinflammation. 15(1): 1-26. Oh WJ, Jung U, Eom HS, Shin HJ, Park HR, (2013) Inhibition of lipopolysaccharide-induced proinflammatory responses by Buddleja officinalis extract in BV-2 microglial cells via negative regulation of NF-kB and ERK1/2 signaling. Molecules. 18(8): 9195-9206. Palak, Soni K, Thakur A, Kohli K (2016) Broccoli: An Insight into Formulation and Patentability Aspects. Drug Des. 5:139. Parker-Athill E, Luo D, Bailey A, Giunta B, Tian J, Shytle RD, Tan J, (2009) Flavonoids, a prenatal prophylaxis via targeting JAK2/STAT3 signaling to oppose IL-6/MIA associated autism. Journal of neuroimmunology. 217(1-2): 20-27. Qi F, Li A, Inagaki Y, Gao J, Li J, Kokudo N, Tang W (2010) Chinese herbal medicines as adjuvant treatment during chemo or radio-therapy for cancer. Bioscience trends. 4(6). Sahranavard S, Ghafari S, Mosaddegh M (2014) Medicinal plants used in Iranian traditional medicine to treat epilepsy. Seizure. 23(5): 328-332. Sammani A, Shammaa E, Chehna F, Rahmo A (2014) The In-Vitro toxic effect of the Glyco alkaloids for Some Solanum Species Against The LIM-1863Cell Line. Pharmacognosy Journal. 6(4). 23-31 Scott O, Galicia-Connolly E, Adams D, Surette S, Vohra S, Yager J Y, (2012) The safety of cruciferous plants in humans: a systematic review. Bio.Med. Research International, 2012. Singh K (2014). Sulforaphane treatment of autism spectrum disorder (ASD). Proceedings of the National Academy of Sciences. 111: 15550-15555. Smith SE, Li J, Garbett K, Mirnics K, Patterson PH (2007) Maternal immune activation alters fetal brain development through interleukin-6. J. Neurosci 27:10695–10702. Song, Mi-Young (2009) Sulforaphane protects against cytokine-and streptozotocin-induced β-cell damage by suppressing the NF-κB pathway. Toxicology and Applied Pharmacology. 57-67. Sharma J, Gairola S, Gaur R.D, Painuli R.M, Siddiqi TO (2013) Ethno medicinal plants used for treating epilepsy by indigenous communities of sub-Himalayan region of Uttarakhand, India. J. Ethnopharmacol.150:353–370. Sharma UK, Pegu S, Hazarika D, Das A, (2012) Medico-religious plants used by the Hajong community of Assam, India. J.Ethnopharmacol. 143:787–800. Tsoi M, Jansen AGM, Bass J, Chiang WC, Seck M, Tsoi V, Wyder P (1998) Excitation of a magnetic multilayer by an electric current. Physical Review Letters, 80(19): 4281. Vargas DL, Nascimbene C, Krishnan C, Zimmerman AW, Pardo CA, (2005) Neuroglial activation and neuroinflammation in the brain of patients with autism. Annals of Neurology: Official Journal of the American Neurological Association and the Child Neurology Society. 57(1): 67-81. Wang SW, Sun YM (2014) The IL-6/JAK/STAT3 pathway: potential therapeutic strategies in treating colorectal cancer. International Journal of Oncology. 44(4): 1032-1040. Winer J, Jung CKS, Shackel I, Williams PM (1999) Development and validation of real-time quantitative reverse transcriptase–polymerase chain reaction for monitoring gene expression in cardiac myocytes in vitro. Analytical biochemistry.270(1): 41-49. Zhang Y, Cho C, Pooner GH, Talalay P (1992) A major inducer of anticarcinogenic protective enzymes from broccoli isolation and elucidation of structure. Proc. Natl. Acad. 2399-2403. Zhang Y, Kensler TW, Cho CG, Posner GR, Talalay P (1994) Anticarcinogenic activities of sulforaphane and structurally related synthetic norbornyl isothiocyanates. Proceedings of the National Academy of Sciences, 91(8), 3147-3150.