The Effect of Erythropoietin on Salpingitis during Ischemia Reperfusion Injury in Rats

  

    
Alteration 
    
95% c.    in. 
    
Reperfusion    time 
    
p-values 
    
 
  
  

    
Wilcoxon
    
glm
  
  

    
without    lesions 0
    
undefined
    
1h
    
1.0000
    
1.0000
  
  

    
without    lesions 0
    
undefined
    
1.5h 
    
1.0000
    
1.0000
  
  

    
without    lesions 0
    
undefined
    
2h 
    
1.0000
    
1.0000
  
  

    
without    lesions 0
    
undefined
    
reperfusion    time
    
1.0000
    
1.0000
  
  

    
without    lesions 0
    
undefined
    
interaction
    
_
    
1.0000
  

Table 4:  The alteration influence of erythropoietin in connection with reperfusion
time.

Discussion

Salpingitis is part of a complex biological response of vascular tissues to harmful stimuli, such as pathogens, damaged cells, or irritants [4]. It is a protective response involving host cells, blood vessels, proteins and other mediators that is intended to eliminate the initial cause of cell injury, as well as the necrotic cells and tissues resulting from the original insult. Along it initiates the process of repair. The classical signs of acute inflammation are pain, heat, redness, swelling, and loss of function. It is a protective attempt by the organism to remove the injurious stimuli and to initiate the healing process. Inflammation is not a synonym for infection, even though the two are often correlated. Inflammation can even occur in absence of infection, although such types of inflammation are usually maladaptive (such as in atherosclerosis). Inflammation is a stereotyped response and therefore it is considered as a mechanism of innate immunity. General chronic salpingitis might lead to a host of diseases, such as hay fever, rheumatoid arthritis and even cancer (as it happens e.g. on cholecystitis for gallbladder carcinoma). Salpingitis can be classified as either acute or chronic. Acute inflammation is the initial response of the body to harmful stimuli and is achieved by the increased movement of plasma and leukocytes (especially granulocytes) from the blood into the injured oviductal tissues. A cascade of biochemical events propagates and matures the inflammatory response, involving the local vascular system, the immune system, and various cells within the injured tissue. Prolonged inflammation, known as chronic inflammation, leads to a progressive shift in the type of cells present at the site of inflammation and is characterized by simultaneous destruction and healing of oviductal tissues from the inflammatory process. Acute inflammation is a short-term process, usually appearing within a few minutes or hours and ceasing upon the removal of the injurious stimulus [5]. It is characterized by the above mentioned five cardinal signs, from which loss of function has multiple causes [6]. These five signs appear [7] when acute inflammation occurs, whereas acute salpingitis may not result in the full set. Pain happens only where the appropriate sensory nerve endings exist in the inflamed area e.g., acute inflammation of endosalpingium does not cause pain unless the inflammation accesses the muscular stratum, which does have pain-sensitive nerve endings. The process of acute inflammation is initiated by cells already present in oviducts, mainly resident macrophages, histiocytes, and mastocytes. These cells present on their endosalpingeal and serosal surfaces, have receptors named Pattern Recognition Receptors (PRRs), which recognize molecules that are broadly shared by pathogens but distinguishable from host molecules, collectively referred to as Pathogen-Associated Molecular Patterns (PAMPs). At the onset of an infection, burn, or other injuries, these cells undergo activation (one of their PRRs recognize a PAMP) and release inflammatory mediators responsible for the above mentioned clinical signs of inflammation. Some of the released mediators such as bradykinin increase the sensitivity to pain (hyperalgesia, dolor). The mediator molecules also alter the blood vessels to permit the migration of leukocytes, mainly neutrophils and macrophages, outside of the blood vessels (extravasation) into the tissue. The neutrophils migrate along a chemotactic gradient created by the local cells to reach the site of injury. The loss of function (functio laesa) is probably the result of a neurological reflex in response to pain. In addition to cell-derived mediators, several acellular biochemical cascade systems consisting of preformed plasma proteins act in parallel to initiate and propagate the inflammatory response. These include the complement system activated by bacteria and coagulation and fibrinolysis systems activated by necrosis, e.g. a burn or a trauma. The acute salpingitis response requires constant stimulation to be sustained. Hence, acute inflammation ceases once the stimulus has been removed. The plasma cascade systems which are activated during acute inflammation are the complement system, the kinin system, the coagulation system and the fibrinolysis system. Specific patterns of acute and chronic salpingitis are seen during particular situations that arise in oviducts, such as when inflammation occurs on an epithelial surface [endosalpigeal or serosal]: granulomatous inflammation, serous inflammation, ulcerative inflammation, but mainly fibrinous or purulent inflammation. Fibrinous inflammation resulting in a large increase in vascular permeability allows fibrin to pass through the blood vessels. If an appropriate procoagulative stimulus is present, such as cancer cells, a fibrinous exudate is deposited. This is commonly seen in serous cavities, where the conversion of fibrinous exudate into a scar can occur between serous membranes, limiting their function. The deposit sometimes forms a pseudomembrane sheet. During inflammation of the endosalpingium, intraoviductal synechiae can be formed. During inflammation of the serosal, pelvic adhesions can be formed. Purulent inflammation resulting in large amount of pus, which consists of neutrophils, dead cells, and fluid. Neisseria gonorrhoeae and Chlamydia trachomatis were originally thought to be the only pathogens that caused acute salpingitis. The immune system is often involved with inflammatory disorders, demonstrated in either allergic reactions or immune system disorders resulting in abnormal inflammation. Non-immune diseases with etiological origins in inflammatory processes include cancer and local ischemic disease. Examples of disorders associated with salpingitis include: autoimmune diseases, auto inflammatory diseases, celiac disease, glomerulonephritis, hypersensitivities, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, transplant rejection, vasculitis and interstitial cystitis.

The following clinical situations show the association between ischemia and female genitalia. Kannan S et al. implicated intrauterine infection which can lead [8] to a fetal inflammatory response syndrome, as one of the causes of perinatal brain injury leading to Periventricular Leukomalacia (PVL) and cerebral palsy. The presence of activated microglial cells has been noted in autopsy specimens of patients with PVL and in models of neonatal hypoxia and ischemia. Intrauterine inflammation leads to activation of microglial cells that may be responsible for the development of brain injury and white matter damage in perinatal period. Braun M et al. demonstrated [9] the suitability of a new in vitro inflammation model of the isolated hemoperfused bovine uterus for the investigation of antiinflammatory substances especially their COX-2 selectivity. Rigor BM Sr classified [10] pelvic cancer causes in several types of pain, i.e., visceral, neuropathic, and somatic pain. Visceral pain is the results of smooth muscles spasms of hallow viscus; distortion of capsule of solid organs; inflammation; chemical irritation; traction or twisting of mesentery and ischemia, or necrosis and encroachment of pelvis and presacral tumors. Dudley DJ supposed [11] an “intrauterine inflammatory response syndrome” that could account for cases of preterm labor in which no infectious organism could be identified, in addition to culture-proven intrauterine infection. There is potential for corticotropin-releasing hormone to regulate inflammatory responses and vice versa.

The inflammatory response must be actively terminated when no longer needed to prevent unnecessary “bystander” damage to oviductal tissues. Failure to do so results in chronic salpingitis and cellular destruction. Resolution of inflammation occurs by different mechanisms in different tissues. Mechanisms that serve to terminate inflammation include: short half-life of inflammatory mediators in vivo [12], production and release of transforming growth factor (TGF) β from macrophages [13-15], production and release of interleukin 10 (IL-10) [16], production of anti-inflammatory lipoxins [17], down regulation of pro-inflammatory molecules, such as leukotrienes, up regulation of anti-inflammatory molecules such as the interleukin 1 receptor antagonist or the soluble Tumor Necrosis Factor Receptor (TNFR), apoptosis of pro-inflammatory cells [18], desensitization of receptors, increased survival of cells in regions of inflammation due to their interaction with the Extracellular Matrix (ECM) [19,20], down regulation of receptor activity by high concentrations of ligands, cleavage of chemokines by Matrix Metalloproteinases (MMPs) might lead to production of anti-inflammatory factors [21], production of resolvins, protectins or maresins. Emerging evidence now suggests that an active, coordinated program of acute inflammation resolution initiates in the first few hours after an inflammatory response begins. After entering in tissues, granulocytes promote the switch of arachidonic acid–derived prostaglandins and leukotrienes to lipoxins, which initiate the termination sequence. Neutrophil recruitment thus ceases and programmed death by apoptosis is engaged. These events coincide with the biosynthesis, from omega-3 polyunsaturated fatty acids, of resolvins and protectins, which critically shorten the period of neutrophil infiltration by initiating apoptosis. As a consequence, apoptotic neutrophils undergo phagocytosis by macrophages, leading to neutrophil clearance and release of anti-inflammatory and reparative cytokines such as transforming growth factor-β1. The anti-inflammatory program ends with the departure of macrophages through the lymphatics [22]. The outcome in a particular circumstance will be determined by the layer in which the injury has occurred and the injurious agent that causes it. The possible outcomes of salpingitis are: resolution, fibrosis, abscess formation or chronic inflammation. However, given some antioxidant capacity of Epo, if it prevents both arachidonic acid-induced and iron-dependent lipid peroxidation, its administration will significantly decreases the salpingitis lesions.

Thus, inflammation is associated with Epo in different tissues. Erbayraktar S et al. [23] established that Epo is a member of the cytokine super family, with significant homology to mediators of growth and inflammation. Results from studies have shown that Epo and its receptor are widely expressed in embryonic and adult tissues, including the central nervous system, gut, kidney, muscles (e.g., smooth, skeletal, and heart), uterus, retina, pancreas, gonads, and lung.

Lappin T [24] supposed that the beneficial effects of hEpo may extend to organs such as ovaries, oviducts, uterus which has Epo receptors. Sasaki R et al. [25] claimed that Epo is both estrogen inducible and produced in oviducts. Masuda S et al. [26] found E2 and hypoxia induced transient, rapidly down regulated stimulation of Epo mRNA in oviductal ampulla and isthmus regions. The E2 action is probably mediated through the E2 receptor and de novo protein synthesis is not required for E2 induced Epo mRNA. Ochiai H et al. [27] attained the synthesis of hEpo protein attempting localized in vivo plasmid DNA gene transfer in laying chicken oviducts.

Conclusion

Epo administration interacted or not with reperfusion tome nonsignificantly short-term altered the salpingitis lesions. 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.

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