Sıçanlarda Asetaminofen Kaynaklı Uterin ve Fallop Tüpleri Hasarına Karşı Eksozomların Kullanımı Üzerine Deneysel Bir Çalışma

Yıl 2023, Cilt: 20 Sayı: 2, 127 - 133, 01.08.2023

Enes Karaman Munevver Baran Zeynep Burcin Gonen Ozge Goktepe , Gozde Ozge Onder Eda Okur Erol Karakas Arzu Yay

https://doi.org/10.32707/ercivet.1332533

Öz

Asetaminofen (APAP), dünya çapında reçetesiz satın alınabilen ve kullanılabilen bir antipiretik ve analjezik ilaçtır. Önerilen maksimum dozdan daha yüksek bir APAP dozu, organ hasarı riskini artırabilir. Mezenkimal kök hücreler (MSC'ler), çeşitli insan dokularından izole edilir ve yararlı etkilerin esas olarak mezenkimal kök hücreden türetilen hüc- re dışı veziküllere (MSC-EV'ler) atfedildiği terapi için kullanılır. Bununla birlikte, hiçbir çalışma eksozomların APAP ile kombinasyon halinde koruyucu etkisine odaklanmamıştır. Bu nedenle bu çalışma APAP toksisitesine karşı eksozom almanın fallop tüpleri ve uterus üzerindeki koruyucu etkisini araştırmak amacıyla yapılmıştır. Kırk dişi Wistar albino sıçan (12-14 haftalık) rastgele dört eşit gruba ayrıldı: kontrol, APAP (1 g/kg APAP aldı), eksozom (30 μg eksozom aldı) ve APAP+eksozom grupları eş zamanlı olarak 1 g/kg APAP ve ardından üç gün sonra kuyruk damarından 30 ug ekso- zom enjeksiyonu yapıldı. Hayvanlar sakrifiye edildikten sonra histolojik ve immünohistokimyasal analizler için uterus ve fallop tüpleri çıkarıldı. Sonuçlar, eksozomların APAP'tan sonra uygulanmasının APAP'ın otofajik etkilerini azalttığını gösterdi. Ayrıca, eksozom tedavisi, otofaji belirteçlerinin (Beclin-1, p62 ve LC3) immünoreaktivite yoğunlukları üzerinde koruyucu bir etki sergiledi. Eksozomlarla yapılan tedavilerin uterus veya fallop tüpleri üzerinde herhangi bir olumsuz etkisi olmamıştır. APAP toksisitesinden sonra eksozomların uygulanması, APAP'ın otofaji etkisi yoluyla hücre ölümünü azaltabilir. Bu bileşiğin APAP'ın toksik etkilerini azaltabileceği öne sürülmektedir. Bu hiperanaljezik etkinin moleküler
mekanizmasını değerlendirmek için ileri çalışmalara ihtiyaç vardır.

Anahtar Kelimeler

Ağrı, asetaminofen, eksozom, otofaji, sıçan

An Experimental Study on the Use of Exosomes against Acetaminophen-induced Uterine and Fallopian Tubes Damage in Rats

Öz

Acetaminophen (APAP) is an antipyretic and analgesic drug that can be bought and used without a prescrip- tion worldwide. A dosage of APAP greater than the maximum recommended dosage can increase the risk of organ damage. Mesenchymal stem cells (MSCs) are isolated from various human tissues and used for therapy, in which beneficial effects are attributed mainly to mesenchymal stem cell-derived extracellular vesicles (MSC-EVs). However, no study has focused on the protective effect of exosomes in combination with APAP. Therefore, the present study was carried out to investigate the protective effect of taking exosomes against APAP toxicity on the fallopian tubes and uterus. Forty female Wistar albino rats (12–14 weeks old) were randomly divided into four equal groups: control, APAP (received 1 g/kg APAP), exosome (received 30 μg of exosomes), and APAP+exosome groups that received simultane- ously 1 g/kg APAP and were followed three days later by a tail vein injection 30 μg of exosomes. The uterus and fallo- pian tubes were removed for histological and immunohistochemical analyses after the animals were sacrificed. The results showed that exosomes' administration after APAP decreased APAP's autophagic effects. Moreover, exosome treatment exhibited a protective effect on the immunoreactivity intensities of autuphagy markers (Beclin-1, p62, and LC3). The treatments with exosomes had no adverse effect on the uterus or fallopian tubes. The administration of exo- somes after APAP toxicity can decrease cell death through the autophagy effect of APAP. It is suggested that this com- pound can decrease the toxic effects of APAP. Further studies are needed to evaluate the molecular mechanism of this hyperanalgesic effect.

Anahtar Kelimeler

Autophagy, acetaminophen, exosome, pain, rat

Kaynakça

• Aleixo JF, Pereira MRF, Montagnini BG, Pereira MJD, Forcato S, Moreira EG, Ceravolo GS, Vieira ML, Kiss ACI, Gerardin DCC. Effect of paracetamol treatment on maternal care and reproductive outcomes in female rat offspring. Reprod Fertil Dev 2020; 32(18): 1311-25.
• Arendrup FS, Mazaud-Guittot S, Jégou B, Kristensen DM. EDC IMPACT: Is exposure during pregnancy to acetaminophen/paracetamol disrupting female reproductive development? Endocr Connect 2018; 7(1): 149-58.
• Baravalia Y, Chanda S. Protective effect of Woodfor- dia fruticosa flowers against acetaminophen- induced hepatic toxicity in rats. Pharm Biol 2011; 49(8): 826-32.
• Bardanzellu F, Neroni P, Dessì A, Fanos V. Parace- tamol in patent ductus arteriosus treatment: effica- cious and safe? Biomed Res Int 2017; 2017: 1438038.
• Börger V, Bremer M, Ferrer-Tur R, Gockeln L, Stam- bouli O, Becic A, Giebel B. Mesenchymal stem/ stromal cell-derived extracellular vesicles and their potential as novel immunomodulatory therapeutic agents. Int J Mol Sci 2017; 18(7): 1450.
• Choi J, Jo M, Lee E, Oh YK, Choi D. The role of au- tophagy in human endometrium. Biol Reprod 2012; 86(3): 70.
• Cohen IV, Cirulli ET, Mitchell MW, Jonsson TJ, Yu J, Shah N, Spector TD, Guo L, Venter JC, Telenti A. Acetaminophen (paracetamol) use modifies the sulfation of sex hormones. EBioMedicine 2018; 28: 316-23.
• Czernek L, Düchler M. Exosomes as messengers between mother and fetus in pregnancy. Int J Mol Sci 2020; 21(12): 4264.
• Dean A, van den Driesche S, Wang Y, McKinnell C, Macpherson S, Eddie SL, Kinnell H, Hurtado- Gonzalez P, Chambers TJ, Stevenson K, Wolfin- ger E, Hrabalkova L, Calarrao A, Bayne RA, Ha- gen CP, Mitchell RT, Anderson RA, Sharpe RM. Analgesic exposure in pregnant rats affects fetal germ cell development with inter-generational re- productive consequences. Sci Rep 2016; 6: 19789.
• Harris DA, Patel SH, Gucek M, Hendrix A, Westbroek W, Taraska JW. Exosomes released from breast cancer carcinomas stimulate cell movement. PloS One 2015; 10(3): e0117495.
• Holm JB, Mazaud-Guittot S, Danneskiold-Samsøe NB, Chalmey C, Jensen B, Nørregård MM, Han- sen CH, Styrishave B, Svingen T, Vinggaard AM, Koch HM, Bowles J, Koopman P, Jégou B, Kristi- ansen K, Kristensen DM. Intrauterine exposure to paracetamol and aniline impairs female reproductive development by reducing follicle reserves and fertility. Toxicol Sci 2016; 150(1): 178-89.
• Johansson HK, Jacobsen PR, Hass U, Svingen T, Vinggaard AM, Isling LK, Axelstad M, Christiansen S, Boberg J. Perinatal exposure to mixtures of endocrine disrupting chemicals reduces female rat follicle reserves and accelerates reproductive ag- ing. Reprod Toxicol 2016; 61: 186-94.
• Kalluri R. The biology and function of exosomes in cancer. J Clin Invest 2016; 126(4): 1208-15.
• Levine B, Kroemer G. Autophagy in the pathogenesis of disease. Cell 2008; 132(1): 27-42.
• Li QC, Liang Y, Su ZB. Prophylactic treatment with MSC-derived exosomes attenuates traumatic acute lung injury in rats. Am J Physiol Lung Cell Mol Physiol 2019; 316(6): L1107-L1117.
• Mehta KJ. Role of iron and iron-related proteins in mesenchymal stem cells: Cellular and clinical as- pects. J Cell Physiol 2021; 236(10): 7266-89.
• Mendt M, Rezvani K, Shpall E. Mesenchymal stem cell-derived exosomes for clinical use. Bone Mar- row Transplant 2019; 54(Suppl 2): 789-92.
• Oestreich AK, Chadchan SB, Popli P, Medvedeva A, Rowen MN, Stephens CS, Xu R, Lydon JP, Demayo FJ, Jungheim ES, Moley KH, Kommagani R. The autophagy gene atg16L1 is necessary for endometrial decidualization. Endocrinology 2020; 161(1): bqz039.
• Parzych KR, Klionsky DJ. An overview of autophagy: morphology, mechanism, and regulation. Antioxid Redox Signal 2014; 20(3): 460-73.
• Patil M. Assessing tubal damage. J Hum Reprod Sci 2009; 2(1): 2-11.
• Ratnasooriya WD, Jayakody JR. Long-term admin- istration of large doses of paracetamol impairs the reproductive competence of male rats. Asian J Androl 2000; 2(4): 247-55.
• Salkın H, Basaran KE. Effects of non-steroidal anti- inflammatory drug (ibuprofen) in low and high dose on stemness and biological characteristics of hu- man dental pulp-derived mesenchymal stem cells. Connect Tissue Res 2023; 64(1): 14-25.
• Sharma P, Diergaarde B, Ferrone S, Kirkwood JM, Whiteside TL. Melanoma cell-derived exosomes in plasma of melanoma patients suppress functions of immune effector cells. Sci Rep 2020; 10(1): 92.
• Tabibzadeh S. The signals and molecular pathways involved in human menstruation, a unique process of tissue destruction and remodelling. Mol Hum Reprod 1996; 2(2): 77-92.
• Timmers L, Lim SK, Hoefer IE, Arslan F, Lai RC, van Oorschot AA, Goumans MJ, Strijder C, Sze SK, Choo A, Piek JJ, Doevendans PA, Pasterkamp G, de Kleijn DP. Human mesenchymal stem cell- conditioned medium improves cardiac function following myocardial infarction. Stem Cell Res 2011; 6(3): 206-14.
• Valadi H, Ekström K, Bossios A, Sjöstrand M, Lee JJ, Lötvall JO. Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 2007; 9(6): 654-9.
• van den Anker JN, Allegaert K. Acetaminophen in the neonatal intensive care unit: shotgun approach or silver bullet. J Pediatr 2018; 198: 10-11.
• Wei Y, Schatten H, Sun QY. Environmental epigenetic inheritance through gametes and implications for human reproduction. Hum Reprod Update 2015; 21(2): 194-208.
• Yin X, Hou T, Huang B, Yang L, Li F. Aptamer recognition-trigged label-free homogeneous electro- chemical strategy for an ultrasensitive cancer- derived exosome assay. Chem Commun (Camb) 2019; 55(91): 13705-8.
• Zhang J, Zhang L, Sun X, Yang Y, Kong L, Lu C, Lv G, Wang T, Wang H, Fu F. Acetylcholinesterase inhibitors for alzheimer's disease treatment amelio- rate acetaminophen-induced liver injury in mice via central cholinergic system regulation. J Pharmacol Exp Ther 2016; 359(2): 374-82.
• Zhao X, Wu D, Ma X, Wang J, Hou W, Zhang W. Exosomes as drug carriers for cancer therapy and challenges regarding exosome uptake. Biomed Pharmacother 2020; 128: 110237.
• Zhou S, Zhao L, Yi T, Wei Y, Zhao X. Menopause- induced uterine epithelium atrophy results from arachidonic acid/prostaglandin E2 axis inhibition- mediated autophagic cell death. Sci Rep 2016; 6: 31408.