The Mechanism of Sesame Oil in Ameliorating Experimental Autoimmune Encephalomyelitis in C57BL/6 Mice

The Mechanism of Sesame Oil in Ameliorating Experimental Autoimmune Encephalomyelitis in C57BL/6 Mice


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علوم پزشکی اراک
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نویسندگان: علی قضاوی , قاسم مسیبی

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نشریه: PHYTOTHERAPY RESEARCH, 4,,3515 - ,

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کد مقاله 1674
عنوان فارسی مقاله The Mechanism of Sesame Oil in Ameliorating Experimental Autoimmune Encephalomyelitis in C57BL/6 Mice
عنوان لاتین مقاله The Mechanism of Sesame Oil in Ameliorating Experimental Autoimmune Encephalomyelitis in C57BL/6 Mice
نوع مقاله بر حسب نگارش پژوهشی اصیل
مقاله برحسب نمایه ISI
IF 1.186
عنوان نشریه PHYTOTHERAPY RESEARCH
نوع نشریه علمی پژوهشی
شماره نشریه 4
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صفحه شروع و پایان در نشریه 3515 -
سال انتشار/ ارائه شمسی
سال انتشار/ارائه میلادی
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چکیده انگلیسیExperimental autoimmune encephalomyelitis (EAE) is a Th1 cell‐mediated autoimmune disease of the CNS that serves as an animal model for multiple sclerosis (MS). The study investigated the effectiveness of treatment with sesame oil on the development of EAE. EAE was induced in 8 week old C57BL/6 mice with an emulsion of MOG35‐55. Therapy with sesame oil (4 mL/kg/day as oral gavage) was started on day 3 before the immunization. IFN‐gamma and IL‐10 production from cultured spleen supernatants were determined by the ELISA method. The results showed that daily oral gavage of sesame oil significantly reduced the clinical symptoms of EAE in C57BL/6 mice. In addition, sesame oil‐treated mice displayed a significantly delayed disease onset. Mononuclear cells isolated from spleen of sesame oil‐treated mice showed a significant decrease in the production of IFN‐gamma compared with control mice (p = 0.001). IL‐10 production was also enhanced in splenic mononuclear cells in sesame oil‐treated mice. The ratio of IFN‐gamma to IL‐10 in sesame oil‐treated EAE mice was significantly less than in non‐treated EAE mice (p = 0.01). This report indicates that sesame oil therapy protected mice from developing EAE by reducing IFN‐gamma secretion. Thus, sesame oil treatment may be effective in MS patients by immunomodulating the Th1 immune response. Copyright © 2011 John Wiley & Sons, Ltd. Keywords: multiple sclerosis; experimental autoimmune encephalomyelitis; sesame oil; cytokines.were started on day 3 before the immunization and continued to day 25 post immunization. Induction of experimental autoimmune encephalomyelitis (EAE). Mice were inoculated s.c. in the flank with 0.1mL of an emulsion containing 200 μg of the encephalitogenic peptideMOG35‐55 (MEVGWYRSPFSRVVHLYRNGK; Diapharm Ltd, Russia) and an equal volume of complete Freund’s adjuvant (Sigma, St Louis, MO) supplemented with 4mg/mL Mycobacterium tuberculosis H37RA (Difco, Detroit, MI, USA). Mice were injected intraperitoneally with 400 ng of pertussis toxin (Sigma) on the day of immunization and 2 days later (Costa et al., 2003). Clinical evaluation of EAE. Following the encephalitogenic challenge, the mice were monitored daily and neurological impairment was scored on an arbitrary clinical score as follows: 0, no clinical sign; 1, partial loss of tail tonicity; 2, complete loss of tail tonicity; 3, flaccid tail and abnormal gait; 4, hind leg paralysis; 5, hind leg paralysis with hind body paresis; 6, hind and foreleg paralysis; 7, moribund or death. The relapse was defined when a mouse developed an increase of the clinical score (more than 1) accompanied by weight loss. Under recommendation of the animal ethics committee, the mice were killed on 25 day post immunization (Mosayebi et al., 2007). Spleen cells preparation. On day 25 post‐immunization, the mice were killed by CO2 inhalation, and the spleens were removed. Spleens were washed in RPMI (modification with 5mM HEPES, 100 U/mL of penicillin and 100 μg/mL of streptomycin, all from Gibco, Life Technologies, Inc., Gaithersburg, MD). The spleens were punctured repeatedly with a pair of forceps to release the spleen cells. Low density mononuclear cells were collected after standard separation on Ficoll‐Paque (Pharmacia Biotech, Uppsala, Sweden), and washed in RPMI with 10% heat‐inactivated fetal bovine serum (FBS). The number of viable cells was assessed by trypan blue exclusion. The cells were then resuspended in RPMI supplemented with 10% FBS, and used for the proliferation assay and cytokine detection. Proliferation of splenocytes. Proliferation was checked by the MTTassay method (Bounous et al., 1992). A total of 3 × 103 cells in 200 μL RPMI 1640 supplemented with 10% FBS were stimulated with 20 μg/mL MOG35‐55 peptide or 5 μg/mL ConA. The plates were then placed in a 5%CO2, 37 °C incubator for 72 h. Twenty microliters of 5mg/mL MTT(3‐(4,5dimethyldiazol‐2‐yl)‐2,5‐dipenyl; Sigma‐Aldrich, St Louis, MO) was added to the cells, followed by incubation for 4 h. After centrifugation, the medium was removed, and 200 μL of DMSO was added to each well. Then, the optical density at 540 nm was measured by microtiter plate reader (Stat Fax2100, USA). The experiments were performed in triplicate sets. Blastogenic responses for the MTT assay were expressed as the mean stimulation index (SI) by dividing the OD values of stimulated cells (C) minus relative cell numbers of unstimulated cells (C) by the relative OD values of unstimulated cells. SI = (C−C)/CMTT. Cytokines assay. Splenocytes isolated from mice on day 25 post‐immunization at a density of 2 × 106 cells/mL were incubated in 1mL cultures in the presence or absence MOG35–55 peptide (20 μg/mL) and placed in a 5% CO2, 37 °C incubator for 96 h. To quantitative IFNgamma and IL‐10 production, supernatants were collected and the amounts of IFN‐gamma and IL‐10 in the supernatants were quantified by ELISA kit (R&D Systems) according to the manufacturer’s protocol. Ninety‐six‐well plates were coated with rat antimouse IFN‐gamma and IL‐10 antibody in coating buffer (pH 9.6) and incubated overnight. After blocking, samples and standards at 1:2 serial dilutions of IFNgamma and IL‐10 (standard curve) were added to the plates and incubated for 60 min at room temperature. After that, biotinylated rat anti‐mouse IFN‐gamma and IL‐10 mAbs were added and incubated for another 60 min. HRP‐conjugated streptavidin were then added for 30 min. After further washings, TMB substrate was added to incubate for 30 min, followed by addition of 0.18mM H2SO4 solution to stop the reaction and reading at 450nm was obtained. Statistical analysis. Statistical analysis was performed with the Mann‐Whitney U‐test to compare the mean maximal score, mean onset day and levels of cytokines. Values of p < 0.05 were considered significant. RESULTS Clinical score The current study investigated the effect of sesame oil administration on evaluation ofEAE (Fig. 1).All animals in the sesame oil‐treated and control groups developed clinical signs of EAE. The onset of the first clinical scores occurred on days 10 and 14 post‐immunization in control group and sesame oil‐treated EAE, respectively (p = 0.02). The mean maximum clinical scores of the sesame oil‐treated group were significantly lower than that in the control group (6 ± 0.7 and 4.8 ± 0.5, respectively, p=0.01). Proliferation assay The study examined the mechanism underlying the amelioration of disease by sesame oil‐treated EAE mice. To examine whether cell proliferation against MOG35‐55 peptide is influenced by sesame oil treatment, mononuclear cells obtained from the spleen of either A. GHAZAVI AND G. MOSAYEBI Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011) sesame oil‐treated or control mice (10 mice in each group) were stimulated with 20 μg/mL of MOG35‐55 peptide or 5 μg/mL ConA. The results show that no significant differences were demonstrated in the cell proliferations between the control and sesame oil‐treated groups (Fig. 2). Cytokine assay The effects of sesame oil on immunomodulation of the immunological response were examined by detection IL‐10 and IFN‐gamma. There was a significant difference between the level of IFN‐gamma production in the culture supernatants of cells from control or sesame oiltreated mice (p = 0.001) (Table 1). The concentration of IL‐10 in supernatants of cells from sesame oil‐treated mice (69 ± 48) was higher than in the control group (58 ± 24) but this difference was not significant. The ratio of IFN‐gamma to IL‐10 in the sesame oil‐treated EAE mice (7.6) was significantly less than in the nontreated EAE mice (16.89) (p = 0.001). DISCUSSION In this study, a protective effect of sesame oil on EAE induced in C57BL/6 mice was demonstrated. Sesame oil significantly delayed the disease onset of EAE and reduced the maximumclinical scores. The mechanism by which sesame oil exhibits its observed beneficial effect is still unknown, but some studies showed that sesame oil has potent antiinflammatory and antioxidant effects (Akimoto et al., 1993; Chavali et al., 2001; Hsu et al., 2005). In addition, sesame oil consumption influences beneficially the blood glucose, lipid peroxidation and antioxidant levels in streptozotocin diabetic rats (Ramesh et al., 2005). Assessment of the cytokine pattern in response to MOG3‐55 re‐stimulation indicated that sesame oil treatment led to a reduction IFN‐gamma secretion as demonstrated for splenocyte cultures. Although immune cells from sesame oil‐treated EAE mice did not show a diminished proliferative response toward MOG35‐55, the levels of IFN‐gamma in the supernatants of splenocytes were lower on sesame oil treatment compared with the control mice, but the levels of IL‐10 in sesame oil‐treated mice were not significantly higher than those in the non‐treatedEAEmice. IFN‐gamma is a pro‐inflammatory cytokine that plays an important role in inflammatory and autoimmune diseases, such as MS (Mana et al., 2006; Popko et al., 1997). The frequencies of IFN‐gamma secreting myelin antigen‐reactive Tcells are increased in blood and especially in the cerebrospinal fluid of MS patients, compared with the controls (Soderstromet al., 1993; Sun et al., 1991). Administration of IFN‐gamma to MS patients exacerbates the disease (Panitch et al., 1985). In addition, lymphocytes from MS Figure 1. Mean clinical course of MOG35‐55‐induced EAE in C57BL/6 mice. Values are means and standard deviations for the 10 mice tested daily in each group. Figure 2. Effects of sesame oil on the proliferation of spleen cells of EAE mice. 25 days after EAE induction, the mononuclear cells obtained from spleen of sesame oil‐treated and non‐treated EAE mice were stimulated with 20 μg/mL MOG35‐55 peptide or 5 μg/mL ConA for 72 h. Increase in cell number was measured by the MTTassay method. Each value represents the mean ± SE (n=10). SESAME OIL AND EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011) patients produce elevated levels of IFN‐gamma in vitro compared with the controls (Hirsch et al., 1985). The protective effect of IFN‐beta therapy in MS patients has been attributed to down‐regulation of IFN‐gamma production (Furlan et al., 2000). There are considerable data demonstrating that under particular conditions and/or in specific tissues IFN‐gamma act to down‐regulate inflammation. Administration of anti‐IFN‐gamma antibody to mice or rats aggravated EAE (Espejo et al., 2001). On a susceptible background, IFN‐gamma‐/‐ mice developed more severe EAE, whereas IFN‐gamma‐/‐ mice on resistant genetic backgrounds become sensitive to EAE (Chu et al., 2000). Animals lacking the gene for IFN‐gamma are much more EAE susceptible (Ferber et al., 1996; Krakowski and Owens, 1996). However, the molecular mechanisms that would explain why in certain situations the antiinflammatory properties of IFN‐gamma remain largely elusive. In addition, histological examination of brain tissues demonstrated reduced levels of infiltration of leukocytes in the sesame oil‐treated EAE mice (Mosayebi et al., 2007). Sesame oil has an antiinflammatory effect and it has been suggested that sesame oil or constituents of sesame oil induce growth arrest and apoptosis of cancer. Recently, it was reported that sesame oil, a vegetable oil enriched with n‐6 polyunsaturated fatty acids attenuated the growth and metastasis of EL4 lymphoma (Salem, 2005). The mechanism of the sesame oil effect on reducing IFN‐γ production and disease intensity is not clear. Some performed studies show that sesame oil causes apoptosis induction in cancer cells. Salem and colleagues showed that prescribing sesame oil prevents the growth and metastasis of enteric EL4 lymphoma in C57BL/6 mice (Salem, 2005). Furthermore, Miyahara and colleagues, by effecting sesamolin on Molt‐4 B range lymphoid cells, showed that sesamolin from sesame seeds induces apoptosis in these cells (Miyahara et al., 2001). So sesame oil may cause a decline in IFN‐γ production and EAE intensity by apoptosis induction or inhibition of inflammatory cell proliferation. CONCLUSION Our study shows, for the first time, that sesame oil reduces the severity of disease and proliferation of leukocytes in EAE mice. The findings suggest that sesame oil might be potentially useful for the treatment of MS. Acknowledgement This work was supported by Research Council of Arak University of Medical Sciences. Conflict of Interest The authors declare that there is no conflict of interest. REFERENCES Adorini L. 2004. Immunotherapeutic approaches in multiple sclerosis. J Neurol Sci 223: 13–24. Akimoto K, Kitagawa Y, Akamatsu Tet al. 1993. Protective effects of sesamin against liver damage caused by alcohol or carbon tetrachloride in rodents. Ann Nutr Metab 37: 218–224. Basso AS, Frenkel D, Quintana FJ et al. 2008. Reversal of axonal loss and disability in a mouse model of progressive multiple sclerosis. J Clin Invest 118: 1532–1543. Bounous DI, Campagnoli RP, Brown J. 1992. Comparison of MTT colorimetric assay and tritiated thymidine uptake for lymphocyte proliferation assays using chicken splenocytes. Avian Dis 36: 1022–1027. Chavali SR, Utsunomiya T, Forse RA. 2001. Increased survival after cecal ligation and puncture in mice consuming diets enriched with sesame seed oil. Crit Care Med 29: 140–143. Chu CQ, Wittmer S, Dalton DK. 2000. Failure to suppress the expansion of the activated CD4 T cell population in interferon gamma‐deficient mice leads to exacerbation of experimental autoimmune encephalomyelitis. J Exp Med 192: 123–128. Costa O, Divoux D, Ischenko A, Tron F, Fontaine M. 2003. Optimization of an animal model of experimental autoimmune encephalomyelitis achieved with a multiple MOG(35‐55) peptide in C57BL6/J strain of mice. J Autoimmun 20: 51–61. Espejo C, Penkowa M, Saez‐Torres I et al. 2001. Treatment with anti‐interferon‐gamma monoclonal antibodies modifies experimental autoimmune encephalomyelitis in interferon‐gamma receptor knockout mice. Exp Neurol 172: 460–468. Ferber IA, Brocke S, Taylor‐Edwards C et al. 1996. Mice with a disrupted IFN‐gamma gene are susceptible to the induction of experimental autoimmune encephalomyelitis (EAE). J Immunol 156: 5–7. Furlan R, Bergami A, Lang R et al. 2000. Interferon‐beta treatment inmultiple sclerosis patients decreases the number of circulating T cells producing interferon‐gamma and interleukin‐4. J Neuroimmunol 111: 86–92. GangurV, Kelly C, Navuluri L. 2005. Sesame allergy: a growing food allergy of global proportions? Ann Allergy Asthma Immunol 95: 4–11; quiz 11–13, 44. Hirsch RL, Panitch HS, Johnson KP. 1985. Lymphocytes from multiple sclerosis patients produce elevated levels of gamma interferon in vitro. J Clin Immunol 5: 386–389. Hsu DZ, Su SB, Chien SP et al. 2005. Effect of sesame oil on oxidative‐stress‐associated renal injury in endotoxemic rats: involvement of nitric oxide and proinflammatory cytokines. Shock 24: 276–280. Huitinga I, van Rooijen N, de Groot CJ, Uitdehaag BM, Dijkstra CD. 1990. Suppression of experimental allergic encephalomyelitis Table 1. IFN‐gamma and IL‐10 production by MNCs obtain from spleen of sesame oil‐treated and non‐treated EAE mice Group No IFN‐gamma (ρg/mL) IL‐10 (ρg/mL) +MOG35‐55 −MOG35‐55 +MOG35‐55 −MOG35‐55 MOG‐EAE+PBS 10 980 ± 42 5 ± 3.7 58 ± 24 26 ± 20 MOG‐EAE+sesame oil 10 527 ± 40 13 ± 12 69 ± 48 24 ± 10 p value 0.001 N.S N.S N.S Data given represent mean ± standard deviation. N.S, not significant. A. GHAZAVI AND G. MOSAYEBI Copyright © 2011 John Wiley & Sons, Ltd. Phytother. Res. (2011) in Lewis rats after elimination of macrophages. J Exp Med 172: 1025–1033. Jan KC, Ho CT, Hwang LS. 2009. Elimination and metabolism of sesamol, a bioactive compound in sesame oil, in rats. Mol Nutr Food Res 53 (Suppl 1): S36–S43. Kang MH, Naito M, Tsujihara N, Osawa T. 1998. Sesamolin inhibits lipid peroxidation in rat liver and kidney. J Nutr 128: 1018–1022. Kanny G, De Hauteclocque C, Moneret‐Vautrin DA. 1996. Sesame seed and sesame seed oil contain masked allergens of growing importance. Allergy 51: 952–957. Krakowski M, Owens T. 1996. Interferon‐gamma confers resistance to experimental allergic encephalomyelitis. Eur J Immunol 26: 1641–1646. Mana P, Linares D, Fordham S, Staykova M, Willenborg D. 2006. Deleterious role of IFNgamma in a toxic model of central nervous system demyelination.AmJ Pathol 168: 1464–1473. Miyahara Y, Hibasami H, Katsuzaki H, Imai K, Komiya T. 2001. Sesamolin from sesame seed inhibits proliferation by inducing apoptosis in human lymphoid leukemia Molt 4B cells. Int J Mol Med 7: 369–371. Mosayebi G, Ghazavi A, Salehi H, Payani MA, Khazae MR. 2007. Effect of sesame oil on the inhibition of experimental autoimmune encephalomyelitis in C57BL/6 mice. Pak J Biol Sci 10: 1790–1796. Mosmann TR, Cherwinski H, Bond MW, Giedlin MA, Coffman RL. 1986. Two types of murine helper T cell clone. I. Definition according to profiles of lymphokine activities and secreted proteins. J Immunol 136: 2348–2357. Oiso N, Yamadori Y, Higashimori N, Kawara S, Kawada A. 2008. Allergic contact dermatitis caused by sesame oil in a topical Chinese medicine, shi‐un‐ko. Contact Dermat 58: 109. Panitch HS, Francis GS, Hooper CJ, Merigan TC, Johnson KP. 1985. Serial immunological studies in multiple sclerosis patients treated systemically with human alpha interferon. Ann Neurol 18: 434–438. Pender MP, Greer JM. 2007. Immunology of multiple sclerosis. Curr Allergy Asthma Rep 7: 285–292. Popko B, Corbin JG, Baerwald KD, Dupree J, Garcia AM. 1997. The effects of interferon‐gamma on the central nervous system. Mol Neurobiol 14: 19–35. Ramesh B, Saravanan R, Pugalendi KV. 2005. Influence of sesame oil on blood glucose, lipid peroxidation, and antioxidant status in streptozotocin diabetic rats. J Med Food 8: 377–381. Rao A, Avni O. 2000. Molecular aspects of T‐cell differentiation. Br Med Bull 56: 969–984. Renno T, Taupin V, Bourbonniere L et al. 1998. Interferon‐gamma in progression to chronic demyelination and neurological deficit following acute EAE. Mol Cell Neurosci 12: 376–389. Sadeghi N, Oveisi MR, Mannan Hajimahmoodi M, Behrooz Jannat B, Mazaheri M, Sadollah Mansouri S. 2009. The contents of sesamol in Iranian sesame seeds. Iran J Pharm Res 8: 101–105. Salem ML. 2005. Systemic treatment with n‐6 polyunsaturated fatty acids attenuates EL4 thymoma growth and metastasis through enhancing specific and non‐specific anti‐tumor cytolytic activities and production of TH1 cytokines. Int Immunopharmacol 5: 947–960. Sankar D, Sambandam G, Ramakrishna Rao M, Pugalendi KV. 2005. Modulation of blood pressure, lipid profiles and redox status in hypertensive patients taking different edible oils. Clin Chim Acta 355: 97–104. Soderstrom M, Link H, Sun JB et al. 1993. T cells recognizing multiple peptides of myelin basic protein are found in blood and enriched in cerebrospinal fluid in optic neuritis and multiple sclerosis. Scand J Immunol 37: 355–368. Sun JB, Olsson T,Wang WZ et al. 1991. Autoreactive Tand B cells responding to myelin proteolipid protein in multiple sclerosis and controls. Eur J Immunol 21: 1461–1468. Trapp BD, Peterson J, Ransohoff RM, Rudick R, Mork S, Bo L. 1998. Axonal transection in the lesions of multiple sclerosis. N Engl J Med 338: 278–285. Utsunomiya T, Chavali SR, ZhongWW, Forse RA. 2000. Effects of sesamin‐supplemented dietary fat emulsions on the ex vivo production of lipopolysaccharide‐induced prostanoids and tumor necrosis factor alpha in rats. Am J Clin Nutr 72: 804–808. SESAME OIL AND EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS Copyright
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