The Effect of Erythropoietin on Ovarian Epithelium Karyorrhexis during Ischemia Reperfusion Injury in Rats

  

    
Decrease 
    
95% c. in
    
Reperfusion time
    
p-values
  
  

    
without lesions -0.1
    
-0.3100922 - 0.1100922
    
1h
    
0.3239
  
  

    
without lesions -0.15
    
-0.371518 - 0.071518
    
1.5h
    
0.1679
  
  

    
without lesions -0.2
    
-0.6201844 - 0.2201844
    
2h
    
0.3239 
  
  

    
without lesions +0.1
    
-0.27768095 – 0.14211844
    
reperfusion time
    
0.4073
  
  

    
without lesions -0.0818182
    
-0.2159977 – 0.0523614
    
interaction
    
0.2246
  

Table 5:  Concise presence of the decreasing influence of erythropoietin in connection with reperfusion time.

Discussion

The following clinical situations show the association between ischemia and ok lesions. Isik S et al. found less karyorrhexis [4] lesions by local antithrombin treatment in hepatic IR injury Wistar rats. Takizawa Y et al. considered [5] that oxidative stress significantly induces DNA peroxidation, apoptotic neuronal death and karyorrhexis 24-72 h after neonatal hypoxic-ischemic (HI) encephalopathy. Sun L et al. found[6] eosinophilic neurons (Ens) with minimally abnormal nuclei and swollen cell bodies at 3 h in the ischemic core and at 12 h in the periphery of post-ischemic gerbils brain. In the ischemic periphery, ENs had slightly atrophic cytoplasm and sequentially developed pyknosis, karyorrhexis and karyolysis over 1 week. Folkerth RD et al. observed [7] nuclear karyorrhexis and/or karyopyknosis with cytoplasmic hypereosinophilia in neurons of the actuate nucleus in consecutive stillbirth brains 22 - 41 gestational weeks old, considering HI lesions such as white matter and brainstem gliosis the cause in part for unexplained stillbirth. Takizawa Y et al. closely associated [8] pontosubicular neuron necrosis and its pathological peculiarity neuronal apoptosis as one of perinatal HI brain injury with presence of karyorrhexis. Hargitai B et al. associated [9] preterm birth with HI encephalopathy including neuronal karyorrhexis mostly at diencephalon and brain stem. Hallak M et al. associated [10] brain injury featured by shrinkage of cells and karyorrhexis at hippocampus and thalamus (P <0.05) with hypoxia and decreased maternal oxygen tension and pH in fetal rats. Tan S et al. induced[11] HI which resulted in significant increase of nitrogen oxides, lipid peroxidation and protein oxidation, with a concomitant decrease of total antioxidant capacity in premature fetal brains rabbit model of acute placental insufficiency in utero. Fetuses delivered 24 h post-ischemia had increased hippocampal nuclear karyorrhexis on histology than controls. Meng SZ et al. manifested [12] neuronal karyorrhexis more predominant in preterm infants with HI basal ganglia necrosis. Fortuna S et al. observed [13] neuronal degeneration and necrosis with nuclear pyknosis and karyorrhexis in a model of mildly HI brain injury. Khera KS et al. noted a pleiotropic karyorrhexis in third or embryonic phase of embryo toxic pathogenesis [14], appeared aggravated, presumably by the preceding second or labyrinthine degeneration of the placental phase in rats embryos. Squier M et al. considered the reactive astrocytosis, macrophage infiltration, karyorrhexis and endothelial swelling or reduplication as criteria [15] for white matter ischemia in early neonatal brains who were stillborn or died due to cerebral palsy. Kalimo H et al. found karyorrhexis and cytorrhexis and removal of their remnants subsequently by macrophages in the great majority of medium-sized neurons of caudate nucleus and putamen [16] after 2-3 days IR injury.

The following situations show the association between Epo and ischemic ovaries. Mahmoodi M et al. found [17] that Epo reduced IR injury and free radical production, increasing follicle survival and function in transplanted ovarian tissue. Sayyah-Melli M et al. determined[18] that rEpo was effective in reducing the oxidative damage of ovarian torsion in operated patients, 18-35 years old, with signs and symptoms of ovarian torsion. Karaca M et al. evaluated [19] the Epo administration as effective in reversing tissue damage induced by IR in ovaries of adult female rats. Suzuki H et al. demonstrated [20] that administration of asialo Epo could effectively enhance the survival of the follicles of transplanted crypreserved ovaries in frozenthawed canine ovarian xenotransplantation. However, David RB et al. did not detect [21] expression of Epo mRNA in porcine ovaries. Kristiansson B et al. concluded [22] that females with carbohydratedeficient glycoprotein syndrome type I have primary ovarian failure, but the syndrome does not affect the terminal charged carbohydrate portion in Epo. Hyttinen JM et al. generated [23] a transgenic calf from in vitro produced bovine embryos microinjected with a gene construct consisting of genomic sequences encoding human Epo. Kamiński M claimed [24] that apoptosis regulates the atrophy of completely developed organs, e.g. thymus, and the hormonal restructuring of ovaries and others but on the other hand, the development of apoptosis is arrested by so called “survival factors” as Epo.

Conclusion

Epo administration interacted or not with reperfusion time nonsignificantly short-term decreased the OK scores. Perhaps, a longer study time than 2 hours or a greater Epo dosage may provide more significant effects.

Acknowledgment

This study was funded by Scholarship by the Experimental Research Center ELPEN Pharmaceuticals (E.R.C.E), Athens, Greece. The research facilities for this project were provided by the aforementioned institution.

References

1. C. Τsompos, C. Panoulis, K. Τοutouzas, G. Ζografos, A. Papalois. The Effect of Erythropoietin on Total Protein Levels during Ischemia Reperfusion Injury in Rats. Int J Neurorehabilitation. 2015; 2: 146.
2. Τsompos C, Panoulis C, Τοutouzas K, Ζografos G, Papalois A. Efecto agudo de la eritropoyetina en los niveles del hematocrito durante una lesión por hipoxia-reoxigenación en ratas. Pren. Méd. Argent. Marzo. 2015; 101: 21-32
3. Osmanağaoğlu MA, Kesim M, Yuluğ E, Menteşe A, Karahan SC. Ovarian-protective effects of clotrimazole on ovarian ischemia/reperfusion injury in a rat ovarian-torsion model. Gynecol Obstet Invest. 2012;74:125-130.
4. Isik S, Tuncyurek P, Zengin NI, Demirbag AE, Atalay F, Yilmaz S, et al. Antithrombin prevents apoptosis by regulating inflammation in the liver in a model of cold ischemia/warm reperfusion injury. Hepatogastroenterology. 2012;59: 453-457.
5. Takizawa Y, Miyazawa T, Nonoyama S, Goto Y, Itoh M. Edaravone inhibits DNA peroxidation and neuronal cell death in neonatal hypoxic-ischemic encephalopathy model rat. Pediatr Res. 2009; 65: 636-641.
6. Sun L, Kuroiwa T, Ishibashi S, Miki K, Li S, Xu H, et al. Two region-dependent pathways of eosinophilic neuronal death after transient cerebral ischemia. Neuropathology. 2009; 29: 45-54.
7. Folkerth RD, Zanoni S, Andiman SE, Billiards SS. Neuronal cell death in the arcuate nucleus of the medulla oblongata in stillbirth. Int J Dev Neurosci. 2008; 26:133-140.
8. Takizawa Y, Takashima S, Itoh M. A histopathological study of premature and mature infants with pontosubicular neuron necrosis: neuronal cell death in perinatal brain damage. Brain Res. 2006; 20;1095:200-206.
9. Hargitai B, Szabó V, Cziniel M, Hajdú J, Papp Z, Szende B, et al. Human brain of preterm infants after hypoxic-ischaemic injuries: no evidence of a substantial role for apoptosis by using a fine-tuned ultrasound-guided neuropathological analysis. Brain Dev. 2004; 26: 30-36.
10. Hallak M, Hotra JW, Kupsky WJ. Magnesium sulfate protection of fetal rat brain from severe maternal hypoxia. Obstet Gynecol. 2000; 96:124-128.
11. Tan S, Zhou F, Nielsen VG, Wang Z, Gladson CL, Parks DA. Sustained hypoxia-ischemia results in reactive nitrogen and oxygen species production and injury in the premature fetal rabbit brain. J Neuropathol Exp Neurol. 1998; 57: 544-553.
12. Meng SZ, Isumi H, Takashima S. Neuropathological characteristics and alteration of the dopamine D2 receptor in hypoxic-ischemic basal ganglia necrosis. Brain Dev. 1998; 20: 98-104.
13. Fortuna S, Pestalozza S, Lorenzini P, Bisso GM, Morelli L, Michalek H. Transient global brain hypoxia-ischemia in adult rats: neuronal damage, glial proliferation, and alterations in inositol phospholipid hydrolysis. Neurochem Int. 1997;31:563-569.
14. Khera KS. A morphologic basis postulated for valproic acid's embryotoxic action in rats. Teratog Carcinog Mutagen. 1992;12: 277-289.
15. Squier M, Keeling JW. The incidence of prenatal brain injury. Neuropathol Appl Neurobiol. 1991;17: 29-38.
16. Kalimo H, Auer RN, Siesjö BK. The temporal evolution of hypoglycemic brain damage. III. Light and electron microscopic findings in the rat caudoputamen. Acta Neuropathol. 1985; 67: 37-50.
17. Mahmoodi M, Soleimani Mehranjani M, Shariatzadeh SM, Eimani H, Shahverdi A. Effects of erythropoietin on ischemia, follicular survival, and ovarian function in ovarian grafts. Reproduction. 2014;147:733-741.
18. Sayyah-Melli M, Rashidi MR, Kaseb-Ganeh M, Rashtchizadeh N, Taghavi S, Ouladsahebmadarek E, et al. The effect of erythropoietin against oxidative damage associated with reperfusion following ovarian detorsion. Eur J Obstet Gynecol Reprod Biol. 2012;162:182-186.
19. Karaca M, Odabasoglu F, Kumtepe Y, Albayrak A, Cadirci E, Keles ON. Protective effects of erythropoietin on ischemia/reperfusion injury of rat ovary. Eur J Obstet Gynecol Reprod Biol. 2009;144:157-162.
20. Suzuki H, Ishijima T, Maruyama S, Yanagimoto Ueta Y, Abe Y, Saitoh H. Beneficial effect of desialylated erythropoietin administration on the frozen-thawed canine ovarian xenotransplantation. J Assist Reprod Genet. 2008; 25:571-575.
21. David RB, Blom AK, Sjaastad OV, Harbitz I. The porcine erythropoietin gene: cDNA sequence, genomic sequence and expression analyses in piglets. Domest Anim Endocrinol. 2001; 20:137-147.
22. Kristiansson B, Stibler H, Wide L. Gonadal function and glycoprotein hormones in the carbohydrate-deficient glycoprotein (CDG) syndrome. Acta Paediatr. 1995;84: 655-659.
23. Hyttinen JM, Peura T, Tolvanen M, Aalto J, Alhonen L, Sinervirta R, et al. Generation of transgenic dairy cattle from transgene-analyzed and sexed embryos produced in vitro. Biotechnology (N Y). 1994;12: 606-608.
24. Kamiński M. Processes of cell necrosis--apoptosis--and their modification by toxic substances. Med Pr. 1994;45: 267-277.