بررسی اثر سویه های بومی پروبیوتیک بر بیومارکر مغزی آمیلوئید بتا در رت‌های آلزایمری شده نژاد ویستار

محورهای موضوعی : فصلنامه زیست شناسی جانوری

احسان اصغری جعفری ¹ (گگروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران)
مریم قبه ² (گروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران)
هانیه جعفری ³ (گروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران)
پریچهره یغمایی ⁴ (گروه زیست شناسی، واحد علوم و تحقیقات تهران، دانشگاه آزاد اسلامی، تهران، ایران)

تاریخ دریافت : 1400/10/16 تاریخ پذیرش : 1400/11/30 تاریخ انتشار : 1401/09/01

کلید واژه: نوروژنز, پلاک‌های آمیلوئیدی, وسترن بلات, آمیلوئید بتا, سویه بومی پروبیوتیک,

چکیده مقاله :

یکی از مهمترین علل پدیده فرسوده شدن جمعیت جهان ، افزایش بیش از اندازه بیماری های نورودژنراتیو است که آلزایمر شایعترین آنها است که با تجمع خارج سلولی پلاک های آمیلوئید بتا (Aβ) در ناحیه هیپوکامپ مشخص می شود. پروبیوتیک ها موجودات ریززنده و فعال میکروسکوپی هستند که استفاده از این میکروارگانیسم ها به منظور حفظ و بهبود سلامت انسان بسیار مهم است. هدف از تحقیق حاضر بررسی اثر دو سویه پروبیوتیک Bifidobacterium longum و Limosilactobacillus reuteri بر بیان ژن Aβ42 و نیز تعداد پلاک‌های آمیلوئیدی و نوروژنز در مغز رت‌های آلزایمری بود. برای این منظور، ابتدا دو سویه پروبیوتیک از ماست سنتی جداسازی، خالص سازی و شناسایی شدند و سپس گروه‌های آزمایشی به این ترتیب دسته بندی شدند: تعداد 30 سر موش نر نژاد ویستار به طور تصادفی به 5 گروه تقسیم شدند. گروه کنترل بدون جراحی و تیمار خاص بودند. سایر گروه‌ها تحت جراحی قرار گرفته و به آنها بتا آمیلوئید تزریق شد. به گروه شم آب (حلال پروبیوتیک) تزریق شد و دو گروه با دوز CFU 109 x 5/2 از دو سویه پروبیوتیک Bifidobacterium longum و Limosilactobacillus reuteri تیمار شدند. در نهایت میزان بیان ژن Aβ42 توسط وسترن بلات و نیز تعداد پلاک‌های آمیلوئیدی و نورون‌ها در مغز رت‌های آلزایمری مورد بررسی قرار گرفتند. در موش های آلزایمری مرگ سلولی نورون‌ها به طور گسترده در هیپوکامپ رخ داد و تعداد پلاک‌های آمیلوئیدی نیز افزایش داشته اند (001/0 p <). همچنین بیان ژن Aβ42 نیز در موش های آلزایمری افزایش داشته است. تیمار موش ها با پروبیوتیک ها تمامی موارد ذکر شده را به طور معناداری بهبود بخشید. دو سویه ی پروبیوتیک Bifidobacterium longum و Limosilactobacillus reuteri احتمالاً می تواند به عنوان یک ماده ی طبیعی مناسب در جهت درمان بیماری آلزایمر و کاهش علائم آن مورد استفاده قرارگیرد که این امر نیازمند انجام تحقیقات بیشتر می باشد.

چکیده انگلیسی:

One of the most important causes of global population decline is the overgrowth of neurodegenerative diseases, including Alzheimer's disease characterized by the extracellular accumulation of amyloid beta (Aβ) plaques in the hippocampus. Probiotics are microscopic organisms that are important for maintaining and improving human health. This study aims at assessing the effect of two probiotic strains, including Bifidobacterium longum and Limosilactobacillus reuteri on Aβ42 gene expression, the count of amyloid plaques, and neurogenesis in the brain of Alzheimer's disease rat models. First, two probiotic strains were isolated, purified and identified from traditional yogurt, and the rats were grouped as follows: thirty male Wistar rats were randomly divided into 5 groups. The control group underwent any surgery with no special treatment. The other groups underwent surgery and after being injected by Aβ42, they were divided into the sham group receiving water (probiotics solvent) and the two experimental groups receiving 2.5 x 109 CFU of Bifidobacterium longum and Limosilactobacillus reuteri strains. Finally, Aβ42 gene expression, the count of amyloid plaques, and neurogenesis were assessed in the brain of Alzheimer's disease rat models. In the brain of induced Alzheimer’s disease rats, neuron death occurred and the number of amyloid plaques increased significantly (p<0.001). Also, Aβ42 gene expression was increased in their brains. Meanwhile, treatment with both probiotic strains notably improved all of these symptoms. Therefore, the use of both Bifidobacterium longum and Limosilactobacillus reuteri probiotic strains seem to be effective in the recovery and treatment of Alzheimer's disease which certainly requires more investigations.

منابع و مأخذ:

1. Ait-Belgnaoui A., Durand H., Cartier C., Chaumlinaz G.,Eutamene H., Ferrier L., 2012. Prevention of gut leakiness by aprobiotic treatment leads to attenuated HPA response to anacute psychological stress in rats. Psychoneurology endocrinology, 37: 1885-1895.


2. Ait-Belgnaoui A., Colom A., Braniste V., Ramalho L., Marrot A., Cartier C., 2014. Probiotic gut effect prevents the chronicpsychological stress-induced brain activity abnormality in mice. Journal of Neurogastroenterology and Motility, 26: 510-520.


3. Akkasheh G., Kashani-Poor Z., Tajabadi-Ebrahimi M., Jafari P., Akbari H.,Taghizadeh M., 2016. Clinical and metabolic response to probioticadministration in patients with major depressive disorder: a randomized,double-blind, placebo-controlled trial. Nutrition, 32: 315-320.


4. Asadi F., Jamshidi A., Khodagholi F., Yans A., Azimi L., Faizi M., 2015. Reversal effects of crocin on amyloid _-induced memory deficit: Modificationof autophagy or apoptosis markers. Pharmacology and Biochemistry, 13(9): 47-58.


5. Ásványi-Molnár N., Sipos-Kozma Z., Tóth Á., Ásványi B., Varga L., 2009. Development of functional dairy food enriched in spirulina (Arthrospira platensis). Tejgazdaság, 69(2):15-22.


6. Bandaru S., Rivenson A., 1993. Inhibitory Effect of Bifidobacterium longum on Colon, Mammary, and Liver Carcinogenesis Induced by 2-Amino-3-methylimidazo[4,5-f]quinoline, a Food Mutagen. Division of  Nutritional carcinogenesis and experimental pathology and toxicology. American Health foundation. Valhalla,  53: 17-193.


7. Bercik P., Park AJ., Sinclair D., Khoshdel A Lu J., Huang X. 2011. The anxiolytic effect of Bifidobacterium longum NCC3001involves vagal pathways for gut-brain communication. Journal of Neurogastroenterology and Motility, 23: 1132-1139.


8. Bravo J.A., Forsythe P., Chew M.V., Escaravage E., Savignac H., Dinan T.G. 2011. Ingestion of Lactobacillus strain regulatesemotional behavior and central GABA receptor expression in a mouse via the vagus nerve. Proceedings of the National Academy of Sciences, 10(8): 16050-16055.


9. Carvalho B., and Abdalla Saad M.J., 2013. Influence of gut microbiota onsubclinical inflammation and insulin resistance. Mediators and Inflammation, 98(6): 734.


10. Cryan JF., Dinan TG., 2012. Mind-altering microorganisms: theimpact of the gut microbiota on brain and behaviour. Nature Reviews Neuroscience, 13: 701-712.


11. D’Andrea MR., Nagele RG., 2006. Targeting the alpha 7 nicotinic acetylcholine receptor to reduce amyloid accumulation in Alzheimer’s disease pyramidal neurons. Current Pharmacy Desisease, 12(6): 677–84.


12. De Simone C., Ciardi A., Grassi A., 1992. Effect Bifidobacterium bifidum and Lactobacillus on gut mucosa peripheral blood B lymphocytes. Immunopharmacology Immunotoxicology, (14): 331-40.


13. Desbonnet L., Garrett L., Clarke G., Bienenstock J., DinanTG., 2008. The probiotic Bifidobacteria infantis: an assessment ofpotential antidepressant properties in the rat. Journal of Psychiatry Research, 43: 164-174.


14. Desbonnet L., Garrett L., Clarke G., Kiely B., Cryan JF., DinanTG., 2010. Effects of the probiotic Bifidobacteria infantis in thematernal separation model of depression. Neuroscience, 1(70): 1179-1188.


15. Desbonnet L., Garrett L., Clarke G., Bienenstock J., Dinan T.G. 2008. The probiotic Bifidobacteria infantis: an assessment ofpotential antidepressant properties in the rat. Journal of Psychiatry Research, (43): 164-174.


16. Dyffy LC., Zielezny MA., Roepenhoff-Talty M., 1994. Effectiveness of Bifidobactruim bidifum in mediating the clinical course of murine rotavirus diarrhea. Pediatry Research, (35): 690-696.


17. Eftekharzadeh B., Ramin M., Khodagholi F., Moradi S., Tabrizian K., Sharif R.., 2012. Inhibition of PKA attenuates memory deficits induced by_-amyloid (1–42), and decreases oxidative stress and NF-B transcription factors. Behavior Brain Research, 226(1): 301–308.


18. Famularo G., Moretti S., Marcellini S., 1997. Stimulation of immunity by probitics. In: Fuller R, ed. Probiotics : Therapeutic an other beneficial effects. London: Chapman and Hall, 13(3): 161.


19. Guarner F., Schaafsma G., 1998. Probiotics. International Journal of Food Microbiology, 39(3): 237-238.


20. HisakoYasui H., Ohwaki M., 1991. Enhancement of Immune Response in Peyer's Patch Cells Cultured with Bifidobacterium breve. Journal of Dairy Science, 74: 1187-1195.


21. S. Hong V., Beja-Glasser B., Nfonoyim A., Frouin S., Li S., Ramakrishnan K., Merry Q., Shi A., Rosenthal B., Barres C., Lemere D., Selkoe B., 2016. Complement and microglia mediate early synapse loss in Alzheimer mouse models. Science, 352: 712-716.


22. Joint F., 2001. WHO Expert consultation on evaluation of health and nutritional properties of probiotics in food including powder milk with live lactic acid bacteria. Córdoba Argentina, 8: 585-615.

23.Kubota M., Kobayashi N., Sugizaki T, Shimoda M., Kawahara M., 2020. Carnosine suppresses neuronal cell death and inflammation induced by 6-hydroxydopamine in an in vitro model of Parkinson's disease. PLOS One, 02(4): 0448

24. Li J., Wang C., Zhang J., Cai J ., Cao Y., Sun X., 2010. Hydrogenrichsaline improves memory function in a rat model of amyloid-betainducedAlzheimer’s disease by reduction of oxidative stress. Brain Research, 13(28): 152–161.


25. Lin M., Beal M., 2006. Mitochondrial dysfunction and oxidative stress in neurodegenerative diseases. Nature, 71(13): 787-795.


26. Mikelsaar M., Zilmer M., 2009. Lactobacillus fermentum ME-3 – an antimicrobial and antioxidative probiotic. Microbial Ecology in Health and Disease, 121(1): 1-27.


27. Masliah E., Mallory M., Hansen L., Richard D., Alford M., Terry R., 1994. Synaptic and neuritic alterations during the progression of Alzheimer’s disease. Neuroscience Letter, 174(1): 67-72.


28. Mesulam M., 2004. The cholinergic lesion of Alzheimer’s disease: pivotal factor or side show? Learning and Memory, 11(1): 43-49.


29. Miettinen M., Vupio-Varkila J., Varkila K., 1996. Production of human tumor necrosis facto alpha, interleukin-6 , and interleukin-10 is induced by lactic acid bacteria. Infect Immun. ASM Journals of Infection and Immunity, 64: 5403-5405.


30. Mohammadi A., Jazayeri S., Khosravi-Darani K., Solati Z., Mohammadpour N.,Asemi Z., 2016. The effects of probiotics on mental health and hypothalamic–pituitary–adrenal axis: a randomized, double-blind, placebo-controlled trial inpetrochemical workers. Nutr. Neuroscience, 19(9): 387-395.


31. Solati Z., Mohammadpour N., Asemi Z. 2016. The effects of probiotics on mental health and hypothalamic–pituitary–adrenal axis: a randomized, double-blind, placebo-controlled trial inpetrochemical workers. Nutr. Neuroscience, 19(9): 387-395.


32. Mohammadi HS., Goudarzi I., Lashkarbolouki T., Abrari K.,Salmani ME., 2014. Chronic administration of quercetin preventspatial learning and memory deficits provoked by chronic stress in rats. Behavior and Brain Research, 2(70): 196-205.


33. Nordberg A., 2001. Nicotinic receptor abnormalities of Alzheimer’s disease: therapeutic implications. Biology and Psychiatry, 49(3): 200-210.

34.Ojetii V.,Petruzziellol C., Minnecol A.,Gnarral A., Gasbarrinll A.,Franceschil F., 2017. Effect of Lactobacillus reuteri (DSM 17938) on methane production in patients affected by functional constipation: a retrospective study. Dermatology, 21: 1702-1708.

35. O’Sullivan E., Barrett E., Grenham S., Fitzgerald P., Stanton C., Ross RP., 2011. BDNF expression in the hippocampus ofmaternally separated rats: does Bifidobacterium breve 6330alter BDNF levels? Beneficial of Microbes, 2: 199-207.


36. Palop J., Mucke L., 2010. Amyloid-β-induced neuronal dysfunction in Alzheimer’s disease: from synapses toward neural networks. Neuroscience, 13(7): 812–8.


37. Qinghui M., Vincent J., 2018. Tavella and Xin M. Luo . Role of Lactobacillus reuteri in Human Health and Diseases. Frontirs of Microbiology, 19(9): 757


38. Quigley E.M., 2010. Prebiotics and probiotics; modifying and mining the microbiota. Pharmacology Research, 61(3): 213–218.


39. Rezaee P., Kasra-Kermanshahi R., Katuli M., 2014. Prebiotics Decrease the Antibacterial Effect of Nano Silver and Nano TiO2 Particles Against Probiotic Bacteria of Food. Current Nutural Food Science, 10(2): 1-6.


40. Salam A., Anatoly Bezkorovainy I., 1993. Survival of bifidobacteria in the presence of bile salt. Journal of the science of food and agriculture, 62(4): 351-354.


41. Sgorbati B., Biavati B., Palenzona D., 1986. Genus bifidobacterium. Bergey's manual of systematic bacteriology. The Genera of Lactic Acid Bacteria, 27(9): 306


42. Schiffrin EJ., Brassart D., Servin A., 1997. Immune modulating of blood leukocytes in humans by lactic acid bacteria : Criteria for strain selection . American Journal of Clinical Nutrition, 66: 15-20.


43. Shariatpanahi M., Khodagholi F., Ashabi G., Khasraghi A., Azimi L.,Abdollahi M. 2015. Ameliorating of memory impairment and apoptosisin amyloid _-injected rats via inhibition of nitric oxide synthase: possibleparticipation of autophagy. Iranian Journal of Pharmaceutical Research, 14(3): 811–824.


44. Athari S., 2018. Lactobacilli and bifidobacteria ameliorate memory and learning deficits and oxidative stress in β-amyloid (1–42) injected rats. Physiology, Nutrition and Metabolism, 43(7): 718-726.


45. Sheehan V., Ross P., Fitzgerald G., 2007. Assessing the acid tolerance and the technological robustness of probiotic cultures for fortification in fruit juices. Innovation of Food Science and Emerging Technology, 8(2): 279-284.


46. Mehrabadi1 S., Sadr S., 2020. Assessment of Probiotics Mixture on Memory Function, Inflammation Markers, and Oxidative Stress in an Alzheimer's Disease Model of Rats. Iranian Biomedical Journal, 24 (4): 220-228


47. Tapia-Arancibia L., Aliaga E., Silhol  M., Arancibia S., 2008. New insightsinto brain BDNF function in normal aging and Alzheimer disease. Brain Research, 59(1): 201-220.


48. Tapia-Arancibia L., Aliaga E., Silhol M., Arancibia S., 2008. New insightsinto brain BDNF function in normal aging and Alzheimer disease. Brain Research, 59(1): 201-220.


49. Mitsuoka T., 1992. The Human Gastrointestinal Tract. The Lactic Acid Bacteria . Springer, 1(69):114.


50. Walsh D., Klyubin I., Fadeeva J., Rowan M., Selkoe D., 2002. Amyloid-β oligomers: their production, toxicity and therapeutic inhibition. Biochemical Society Trans, 30(4): 552-557.


51. Wang Y., Kasper L.H., 2014. The role of microbiome in central nervoussystem disorders. Brain and Behavioral Immunity, 1(38): 1-12.


52. Wang Y., Wu Y., Wang Y., Xu H., Mei X., Yu D., 2017. Antioxidant properties of probiotic bacteria. Nutrients, 1(9): 521.


53. WennLiu Y., HsienLiu W., ChenWu C., ChenJuan Y., ChenWu Y., PingTsai H., Wang S., ChiehTsai Y., 2016. Psychotropic effects of Lactobacillus plantarum PS128 in early life-stressed and naïve adult mice. Brain Research, 16(15): 1-12.