|Year : 2016 | Volume
| Issue : 1 | Page : 24-28
The effect of the antioxidant drug "U-74389G" on salpingitis during ischemia-reperfusion injury in rats
Constantinos Tsompos1, Constantinos Panoulis2, Konstantinos Tutouzas3, Aggeliki Triantafyllou4, George Zografos3, Apostolos Papalois5
1 Department of Obstetrics and Gynecology, Mesologi County Hospital, Etoloakarnania, Greece
2 Department of Obstetrics and Gynecology, Aretaieion Hospital, Athens University, Pikermi, Attica, Greece
3 Department of Surgery, Ippokrateio General Hospital, Athens University, Pikermi, Attica, Greece
4 Department of Biologic Chemistry, Athens University, Pikermi, Attica, Greece
5 Experimental Research Centre, ELPEN Pharmaceuticals, S.A. Inc., Co., Pikermi, Attica, Greece
|Date of Submission||02-Jan-2016|
|Date of Acceptance||05-Apr-2016|
|Date of Web Publication||16-Jun-2016|
Department of Obstetrics and Gynecology, Mesologi County Hospital, Nafpaktou Street, Mesologi 30200, Etoloakarnania
Source of Support: None, Conflict of Interest: None
Objective: This experimental study examined the effect of the antioxidant drug "U-74389G," on rat model and particularly in an oviductal ischemia-reperfusion (IR) protocol. The probable beneficial effect of that molecule was studied pathologically using mean salpingitis (S) lesions.
Materials and Methods: Forty rats of mean weight 231.875 g were used in the study. S lesions were evaluated at 60 min of reperfusion (Groups A and C) and 120 min of reperfusion (Groups B and D), A and B without but C and D with the U-74389G administration.
Results: U-74389G administration nonsignificantly altered the S scores without S lesions by 0 (P = 1.0000). Reperfusion time nonsignificantly altered the S scores without S lesions by 0 (P = 1.0000). However, U-74389G administration and reperfusion time together nonsignificantly altered the S scores without S lesions by 0 (P = 1.0000).
Conclusions: U-74389G administration whether it interacted or not with reperfusion time nonsignificantly altered without lesions the salpingitis lesions within short-term time context of 2 h. Perhaps, a longer study time than 2 h or a higher drug dose may provide significant effects.
Keywords: Ischemia, reperfusion, salpingitis, U-74389G
|How to cite this article:|
Tsompos C, Panoulis C, Tutouzas K, Triantafyllou A, Zografos G, Papalois A. The effect of the antioxidant drug "U-74389G" on salpingitis during ischemia-reperfusion injury in rats. J Curr Res Sci Med 2016;2:24-8
|How to cite this URL:|
Tsompos C, Panoulis C, Tutouzas K, Triantafyllou A, Zografos G, Papalois A. The effect of the antioxidant drug "U-74389G" on salpingitis during ischemia-reperfusion injury in rats. J Curr Res Sci Med [serial online] 2016 [cited 2019 Sep 18];2:24-8. Available from: http://www.jcrsmed.org/text.asp?2016/2/1/24/184125
| Introduction|| |
Permanent or transient damage with serious implications on adjacent organs and certainly on patients' health may be due to tissue ischemia and reperfusion (IR). Although important progress has been made regarding the usage of U-74389G in managing these kinds of damages, satisfactory answers have not been given yet to fundamental questions, such as, at what velocity this factor acts, when should it be administered and at what dosage. The particularly satisfactory action of U-74389G as an antioxidant agent has been noted in several performed experiments. However, just a few reports were found concerning the U-74389G trial in IR experiments, not completely covering this particular matter. Furthermore, numerous publications have addressed trials of similar antioxidant molecules U-74389G or better, 21-(4- [2,6-di-1-pyrrolidinyl-4-pyrimidinyl]-1-piperazinyl)- pregna-1, 4, 9 (11)-triene-3,20-dione maleate salt  is an antioxidant which prevents both arachidonic acid-induced and iron-dependent lipid peroxidation. It protects against IR injury in animal heart, liver and kidney models. These membrane-associating antioxidants  are particularly effective in preventing permeability changes in brain microvascular endothelial cells monolayers. A meta-analysis of 14 published seric variables, coming from the same experimental setting, tried to provide a numeric evaluation of U-74389G efficacy at the same endpoints [Table 1]. 
|Table 1: The U-74389G influence±standard deviation on the levels of some seric variables concerning reperfusion time|
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The aim of this experimental study was to examine the effect of U-74389G on rat model and particularly in an oviductal IR protocol. The beneficial effect or noneffectiveness of that molecule were studied by evaluating mean salpingitis (S) lesions. Salpingitis is the aseptic inflammation of oviducts induced by IR.
| Materials and methods|| |
This experimental study was licensed by Veterinary Address of East Attiki Prefecture under 3693/November 12, 2010 and January 14/10, 2012 decisions. All consumables, equipment and substances, were a courtesy of Experimental Research Centre of ELPEN Pharmaceuticals Co. Inc., S.A. at Pikermi, Attiki. Accepted standards of humane animal care were adopted for albino female Wistar rats. Pre-experimental normal housing in the laboratory for 7 days included ad libitum diet. Post-experimental awakening and preservation of the rodents was not permitted, even if euthanasia was needed. They were randomly delivered to four experimental groups with 10 animals in each one. Ischemia for 45 minutes was followed by reperfusion for 60 min (Group A). Ischemia for 45 min was followed by reperfusion for 120 min (Group B). Ischemia for 45 min was followed by immediate U-74389G intravenous (IV) administration and reperfusion for 60 min (Group C). Ischemia for 45 min was followed by immediate U-74389G IV administration and reperfusion for 120 min (Group D). The molecule U-74389G dosage was 10 mg/kg body weight of the animals.
The detailed pre-narcotic and general anesthesiologic techniques for animals are described in the related reference.  Continuous intra-experimental oxygen supply, electrocardiogram, and acidimetry were provided.
The protocol of IR was followed. Ischemia was caused by laparotomic forceps clamping the inferior aorta over renal arteries for 45 min. Reperfusion was induced by removing the clamp and reestablishment of inferior aorta patency. The drug was administered at the time of reperfusion through catheterized inferior vena cava. The S lesions evaluations were performed at 60 min of reperfusion (for Groups A and C) and at 120 min of reperfusion (for Groups B and D). Forty female Wistar albino rats were used of mean weight 231.875 g (standard deviation [SD]: 36.59703 g), with min weight ≥165 g and max weight ≤320 g. Rats' weight could be potentially a confusing factor, for example, the more obese rats could have higher or less S scores. Furthermore, a detailed pathological study  and grading of S findings was performed by the Pathology Department of Athens University Medical School. The scores were defined as: 0, lesions not found; 1, mild lesions found; 2, moderate lesions found; and 3, severe lesions found. The previous grading was transformed as follows: (0-0.499) without lesions, (0.5-1.499) mild lesions, (1.5-2.499) moderate lesions and (2.5-3) severe lesions, because the study concerned score ranges rather than point scores.
Model of ischemia reperfusion injury
Twenty control rats of mean weight 252.5 g (SD: 39.31988 g) experienced ischemia for 45 min followed by reperfusion.
Reperfusion which lasted 60 min in 10 control rats of mean weight 243 g (SD: 45.77724 g] and mean without S lesions score 0 (SD: 0) [Table 2].
|Table 2: Weight and salpingitis score mean levels and standard deviation of groups|
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Reperfusion which lasted 120 min in 10 control rats of mean weight 262 g (SD: 31.10913 g) and mean without S lesions score 0 (SD: 0) [Table 2].
Lazaroid (test) group
Twenty rats of mean weight 211.25 g (SD: 17.53755 g) experienced ischemia for 45 min followed by reperfusion at the beginning of which 10 mg U-74389G/kg body weight were IV administered.
Reperfusion which lasted 60 min in 10 test rats of mean weight 212.5 g (SD: 17.83411 g) and mean without S lesions score 0 (SD: 0) [Table 2].
Reperfusion which lasted 120 min in 10 test rats of mean weight 210 g (SD: 18.10463 g) and mean without S lesions score 0 (SD: 0) [Table 2].
Every weight group was compared to each of the three remaining groups by applying the paired t-test [Table 3]. Any emerging significant difference among S scores was investigated as to whether it could be accounted for by the weight correlations. Furthermore, every S scores group was compared to each of the three remaining groups by applying Wilcoxon signed-rank test [Table 3]. The application of generalized linear models (glm) with S scores as the dependent varable was followed. The three independent variables were withholding/administration of U-74389G, reperfusion time, and their interaction. Insertion of the rats' weight as an independent variable in the glm analysis resulted in a nonsignificant relation (P=1.0000), so further investigation was not needed.
|Table 3: Statistical significance of mean values difference for groups after statistical paired t-test application for weight and Wilcoxon signed-rank test for scores|
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| Results|| |
Glm resulted in: U-74389G administration nonsignificantly altered the S scores without S lesions by 0 (P = 1.0000). This finding was in accordance with the results of Wilcoxon signed-rank test (P = 1.0000). Reperfusion time nonsignificantly altered the S scores without S lesions by 0 (P = 1.0000), also in accordance with the Wilcoxon signed-rank test (P = 1.0000). However, U-74389G administration and reperfusion time together nonsignificantly altered the S scores without S lesions by 0 (P = 1.0000). Reviewing the above and [Table 3] and [Table 4] sum up the altering influence of U-74389G on reperfusion time.
|Table 4: The alteration influence of U-74389G in connection with reperfusion time|
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| Discussion|| |
Salpingitis is part of a complex biological response of vascular tissues to harmful stimuli, such as pathogens, damaged cells, or irritants.  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, and to initiate the process of repair. Inflammation is not a synonym for infection, even though the two are often correlated. Inflammation can even occur in the absence of infection although such types of inflammation are usually maladaptive (such as in atherosclerosis). Inflammation is a stereotyped response, and therefore, is considered as a mechanism of innate immunity. General chronic inflammation 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). 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.  It is characterized by the five cardinal signs of pain, heat, redness, swelling, and loss of function, among which loss of function has multiple causes.  These five signs appear  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 certain 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, for example, 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 is 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 results in a large amount of pus, which consists of neutrophils, dead cells, and fluid. Neisseria More Details gonorrhoeae and Chlamydia trachomatis were originally thought to be the only pathogens that caused acute salpingitis. The immune system is often involved in inflammatory disorders, demonstrated in either allergic reactions or immune system disorders resulting in abnormal inflammation. Nonimmune diseases with etiological origins in inflammatory processes include cancer and local ischemic disease. Examples of disorders associated with salpingitis include autoimmune diseases, autoinflammatory diseases, celiac disease, glomerulonephritis, hypersensitivities, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, transplant rejection, vasculitis and interstitial cystitis.
Ischemia may be associated with female genitalia. Kannan et al. implicated intrauterine infection which can lead  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 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 the perinatal period. Braun and Kietzmann demonstrated  the suitability of a new in vitro inflammation model of the isolated hemoperfused bovine uterus for the investigation of anti-inflammatory substances, especially their COX-2 selectivity. Rigor observed that  pelvic cancer results in several types of pain, i.e. visceral, neuropathic, and somatic pain. Dudley supposed  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 the 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. The anti-inflammatory program ends with the departure of macrophages through the lymphatics.  The outcome in a particular circumstance will be determined by the layer in which the injury has occurred and the injurious agent that is causing it. The possible outcomes to salpingitis are: resolution, fibrosis, abscess formation or chronic inflammation. However, if U-74389G prevents both arachidonic acid-induced and iron-dependent lipid peroxidation, its administration will significantly decrease the salpingitis lesions.
Haddad et al. implicated  free radicals as contributors in the development of inflammation under related conditions. Streptococcus pneumoniae nisms were inoculated into the right tympanic cavity; sterilized phosphate-buffered saline solution was injected into the left ear to serve as a control in guinea pigs. The animals were given intraperitoneal injections of a lazaroid U-74389G compound 40 mg/kg or its vehicle every 12 h. Middle ear mucosa was collected and used for the assay. The lazaroid significantly (P < 0.05) suppressed production of lipid hydroperoxide of the middle ear mucosa with acute otitis media for up to 24 h. These results suggest that lazaroids may reduce lipoperoxidation in the middle ear at an early stage of acute otitis media.
| Conclusions|| |
U-74389G administration whether or not it interacted with reperfusion time nonsignificantly altered the salpingitis scores without salpingitis lesions within a short period of time (2 h). At present, it has proved ineffective for S. However, a longer study time than 2 h or a higher drug dose may provide more significant effects.
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.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
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[Table 1], [Table 2], [Table 3], [Table 4]