How do red and infrared low-level lasers affect folliculogenesis cycle in rat’s ovary tissue in comparison with clomiphene under in vivo condition

Tabriz University of Medical Sciences, 5/89/137Peer reviewedPublisher PD

Investigation on the effects of Melatonin on the thyroid gland in pregnant and infant rats subjects to electromagnetic radiation

Gebelik Dönemminde Elektromanyetik Alana Maruz Bırakılan Ratlarda ve Yavrularda Melatonin'in Tiroid Bezi Üzerine Etkilerinin İncelenmesi Bu çalışmada, günlük yaşamda sık karşılaşılan 50 Hz titreşimli elektromanyetik alanın (EMA), gebelik döneminde anne ve yavru sıçan tiroid bezi üzerine etkilerine ve bu etkilere melatoninin koruyucu etkisi olup olmadığıa araştırıldı. Çalışmamızda östrus siklus takibi ardından, gebe kalması sağlanan 28 Wistar albino rat rasgele seçilerek 4 gruba (her grup 7 denek) ayrıldı. Grup I: kontrol, Grup II: sham EMA cihazı çalıştırılmadan denekler 4 saat boyunca bu cihazın içinde tutuldu + Melatonin verildi, Grup III: EMA'a maruz kalan, Grup IV: EMA'a maruz kalan + Melatonin verilen grup. Sıçanlar gebeliğin 8. gününden doğum yapana kadar her gün 4 saat boyunca EMA'a (3 mikT, 50Hz) maruz bırakıldı. Melatonin gebeliğin 8'inci gününden itibaren her gün (grup II ve IV) gündüz saat 10'da 4 mg/kg ( 0.1 ml/100 g) intraperitoneal (IP) olarak verildi. Melatonin verilmeyen gruplara ise aynı zamanda fizyolojik salin (0.1 ml/100 g) %5 etanol ip olarak verildi. Doğum sonrası, anne sıçanlar ve yeni doğan yavruların rutin histolojik takipten sonra tiroid dokuları alınarak H&E, Mallory anilin Blue, PAS ve immunhistokimya (TUNEL ve Caspase 3) ile boyanarak histolojik değerlendirme ve morfometrik ölçümler yapıldı. Ayrıca ultrastrukturel düzeydeki değişiklikler, transmissiyon elektron mikroskobu ile değerlendirildi. İmmunhistokimya (TUNEL ve Caspase 3) değerlendirmeye göre EMA'a maruz kalan anne ve yavru sıçanlarda hücresel hasarın ve apoptozun artmış olduğu gözlendi. Histopatolojik ve morfometrik değerlendirmeye göre, EMA'a maruz kalan sıçanların, kontrol, sham ve EMA'a maruz kalan + Melatonin verilen gruplara göre TUNEL pozitif ve hasarlı hücre sayısının artması, tiroid follikül çap ölçümünde follikül çapların azalması, intrafoliküller kollidin azalması, , interfolliküler bağ dokusunun hacminin artması gibi bulgular saptandı. Tiroid follikül hücrelerin ultrastrukturel analizinde EMA'a maruz kalan grupta, yoğun heterokromatin ve düzensiz çekirdek, sitoplazmada kolloid damlacıkların oluşumu, rER'mda dilatasyon ve silindirikli mitokondriler gibi değişiklikler gözlendi. Bu değişikler EMA'a maruz kalan + Melatonin alan grupta az olarak saptandı. Statiksel sonuçların değerlendirmesinde EMA'a maruz kalan + Melatonin verilen grup ile EMA'a maruz kalan grubun arasında tamamen anne ratlarda ve kısmen yavru ratlarda belirgin bir fark olduğu bulundu. Sonuç olarak EMA'a bağlı oluşan hasara karşı melatonin'in antioksidan etkisinin, tiroid bezi üzerinde koruyucu etkiye sahip olduğunu düşünmekteyiz. Anahtar kelime: Elektromanyetik Alan, Tiroid bez, Melatonin, İmmunhistokimya Investigation on the Effects of Melatonin on the Thyroid Gland in Pregnant and Infant Rats Subjects to Electromagnetic Radiation. In this study, we investigated protective effect of melatonin on thyroid glands in pregnancy period and infant rats subjects to 50 Hz frequents electromagnetic radiation that is frequently encounter in daily life. After monitoring estrous cycle the rats were mated and tested for gestation then 28 Wistar pregnant rats randomly selected and put into 4 groups (each group n = 7 ): Group I control, Group II sham (keeping rats in EMF apparatus for 4 hours without any exposure to EMF + Melatonin ), Group III EMF exposed and group IV EMF exposed + Melatonin. The EMF exposed and sham groups (II and IV groups) had intraperitoneal singe dose of a 4 mg/kg melatonin (0.1 ml/100 g) at 10:00 o'clock daily up to the end of gestation. For EMF exposed and control groups had daily intraperitoneal injections of physiologic saline (0.1 ml/100 g) containing 5% ethanol. After birth of rats, mother rats and their infants of thyroid tissues were submitted for histological examination under light microscopy by ımmunohistochem (TUNEL ve Caspase 3), H&E, Mallory anilin Blue, PAS staining methods and morphometric assays. Also ultrastructural changes were assessed by transmission electron microscope. According to immunohistochem examinations (TUNEL and Caspase 3) in EMF exposed animals were observed cellular damage and apoptosis. Histopathological and morphometric analysis showed a significantly increased number cellular damage and apoptotic cells, decreased diameter assay of thyroid follicles, decreased intrafollicular colloid content in lumen, increased volume of interfollicular connective tissue and in EMF exposed animals compared to EMF exposed + Melatonin, control and sham non-exposed groups. Ultrastructural analysis of thyroid follicular cells in the EMF exposed group showed alterations in mitochondria as slender form, dilatation of granular endoplasmic reticulum, numerous apical lysosomes, finds of several colloid droplets dentate form of cell nuclei with incisions and a higher density of heterochromatin compared to the other groups were also observed. There were statistically significant differences between EMF exposed animals compared to EMF exposed + Melatonin animals groups. Finally results demonstrate that melatonin antioxidant benefits may have a preventive effect against EMF exposed injury on thyroid glands. Key words: Electromagnetic Fields, Thyroid gland, Melatonin, Immunohistochemistr

Antioxidant effect of genistein on ovarian tissue morphology, oxidant and antioxidant activity in rats with induced polycystic ovary syndrome

Background: Oxidative stress is the most frequent cause of female infertility disorders including polycystic ovary syndrome (PCOS). Genistein as a major component of soybean isoflavone scavenges free radicals by antioxidant activities. Objective: The present study examines the antioxidant effects of genistein on ovarian tissue following experimental PCOS in rats. Materials and Methods: Twenty female Wistar rat were randomly divided into the following groups (n=5 each group): (I) control group (no treatment); (II) induced PCOS (injection of estradiol valerate); (III) genistein-treated non-PCOS (received genistein); and (IV) genistein-treated PCOS groups. The weight of rats were measured and the blood samples collected and centrifuged. The oxidant and antioxidant activity of plasma and ovaries were measured. All rats were sacrificed under anesthesia, and ovaries were collected and weighted. Histological examination and follicular qualitywere assessed by staining. Results: In histological observation, the induced PCOS rats displayed more number of atretic follicles and the follicular quality in genistein-treated rats was similar to the control groups. The plasma and ovaries malondialdehyde levels significantly increased in PCOS rats (p < 0.001), while the total antioxidant capacity levels, glutathione peroxidase, and superoxide dismutase activities significantly decreased (p < 0.001). The plasma and ovary  malondialdehyde levels significantly decreased in PCOS rats that were treated with genistein (p < 0.001) and the total antioxidant capacity (p < 0.05), glutathione peroxidase, and superoxide dismutase activities significantly increased (p < 0.001). Conclusion: Treatment with genistein preserved follicular quality by increasing antioxidant activities and scavenging oxidant levels in PCOS rats. Key words: Genistein, Antioxidant, Ovary, Polycystic, Follicle, Rats

The Anti-Inflammatory Effect of Erythropoietin and Melatonin on Renal Ischemia Reperfusion Injury in Male Rats

Purpose: Renal ischemia reperfusion (IR) is an important cause of renal dysfunction. It contributes to the development of acute renal failure (ARF). The purpose of this study was to investigate the anti-inflammatory effect of erythropoietin (EPO) and melatonin (MEL), which are known anti-inflammatory and antioxidant agents, in IR-induced renal injury in rats. Methods: Male Wistar Albino rats were unilaterally nephrectomized and subjected to 45 min of renal pedicle occlusion followed by 24 h reperfusion. MEL (10mg/kg, i.p) and EPO (5000U/kg, i.p) were administered prior to ischemia. After 24 h reperfusion, blood samples were collected for the determination of total antioxidant capacity (TAC), malondialdehyde (MDA) and serum creatinine levels. Also, renal samples were taken for Immunohistochemical evaluation of Bcl2 and TNF-α (tumor necrosis factor-α) expression. Results: Ischemia reperfusion increased creatinine, TAC, MDA levels and TNF-α expression, also, IR decreased Bcl2 expression. Treatment with EPO or MEL decreased creatinine, MDA levels, and increased TAC level. Also, MEL up-regulated Bcl2 expression and down-regulated TNF-α expression compared with EPO. Conclusion: Treatment with EPO and MEL had a curative effect on renal IR injury. These results may indicate that MEL protects against inflammation and apoptosis better than EPO in renal IR injury

Troxerutin protects hippocampal neurons against amyloid beta-induced oxidative stress and apoptosis

Alzheimer’s disease (AD) is an age-related neurodegenerative disease linked with increased production and/or deposition of amyloid-beta (Aβ) in the brain. The aim of the present study was to investigate the possible neuroprotective effect of troxerutin on an animal model of Alzheimer's disease. Alzheimer model was induced by a single dose intracerebroventricular (ICV) injection of Aβ 1–42 (5 nmol/5 μl). Thereafter, troxerutin (300 mg/kg) was gavaged for 14 days. The hippocampal malondialdehyde (MDA) levels and enzymatic activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), and acetylcholinesterase (AChE) were measured using enzymelinked immunosorbent assay (ELISA) method. In addition, the number of apoptotic cells in the dentate gyrus (DG) was assessed by TUNEL kit. The results showed that ICV microinjection of Aβ 1-42 increased MDA levels, reduced SOD and GPx, and increased AChE activities in the hippocampus. Chronic administration of troxerutin significantly attenuated MDA levels and AChE activity and increased SOD and GPx activities in the hippocampus. Moreover, the number of apoptotic cells was decreased by troxerutin treatment. Taken together, our study demonstrated that troxerutin could increase the resistance of hippocampal neurons against apoptosis, at least in part, by diminishing the activity of AChE and oxidative stress. Therefore, troxerutin may have beneficial effects in the management of Alzheimer's disease

The Effects of 50 Hz Electromagnetic Fields Induction of Apoptosis in Rat Ovarian Follicles

Objective: The increases application of electromagnetic fields (EMFs) equipments in last decades led to some diseases. Therefore, many studies focused in this issue mainly on critical organs like central nervous system and reproductive function. In the meantime, female infertility by EMFs is one of the most important challenges in current studies. Materials and Methods: Thirty rats were selected randomly and divided into 3 groups (n = 10). Group 1 received 50 Hz EMFs for 8 weeks (3 weeks via intrauterine + 5 weeks after births). Group 2 received 50 Hz EMFs for 13 weeks (3 weeks via intrauterine + 10 weeks after birth). Group 3 was considered as a control in the normal condition. After this period, evaluation apoptosis in ovarian follicles were analyzed by TUNEL method. Results: The results of this study show a significantly increased of apoptotic cell members in experimental groups. Degenerative and pathologic changes were found in ovarian tissue and follicles besides which indicates increase the number of apoptotic cells in experimental groups. As well as, this study has shown that oocyte nucleus was smaller and deformed and apoptotic vacuoles increased in granulose cell. Conclusion: According to the results, we observed that EMFs can changes oogenesis cycle through destructive impacts on ovary structure and function. Of course, this problem depends on frequency and duration of exposure to these waves

Therapeutic potential of genistein in ovariectomy-induced pancreatic injury in diabetic rats: The regulation of MAPK pathway and apoptosis

Objective(s): Genistein, as a phytoestrogen found in legumes, has several biological activities in general and anti-diabetic activity particularly. In this study, we investigated the effect of genistein on proteins involved in β-cell proliferation, survival and apoptosis to further reveal its anti-diabetic potential in the ovariectomized diabetic rat. Materials and Methods: We used three-month-old female Wistar rats that either underwent ovariectomy (OVX) or received a sham surgery (Sham). In a subsequent series of experiments, OVX rats received high-fat diet and low dose STZ to induce diabetes (OVX.D) and genistein treatment (OVX.D.G). Western blot analysis was used for the assessment of phosphorylation of ERK1/2 and AKT and expression of Bcl-2 and caspase-3 in pancreas tissue. Hematoxylin-Eosin (H&E) staining was used for histopathological assessment. Results: Genistein induced AKT and ERK1/2 phosphorylation protein expression of Bcl-2 in the pancreas. In addition, genistein suppressed protein level of caspase-3. Administration of genistein significantly improved hyperglycemia in ovariectomized diabetic rat, concomitant with improved islet β-cell morphology and mass. Conclusion: These findings suggest that the beneficial antidiabetic effect of genistein partially mediated by directly modulating pancreatic β-cell function via activation of the AKT, ERK1/2, and Bcl-2, as cell survival and anti-apoptotic factors, and decreasing of proapoptotic caspase-3

Combination of Vildagliptin and Ischemic Postconditioning in Diabetic Hearts as a Working Strategy to Reduce Myocardial Reperfusion Injury by Restoring Mitochondrial Function and Autophagic Activity

Purpose: Diabetic hearts are resistant to cardioprotection by ischemic-postconditioning (IPostC). Protection of diabetic hearts and finding related interfering mechanisms would have clinical benefits. This study investigated the combination effects of vildagliptin (Vilda) and IPostC on cardioprotection and the levels of autophagy and mitochondrial function following myocardial ischemia/reperfusion (I/R) injury in type-II diabetic rats. Methods: Diabetes was established by high fat diet/low dose of streptozotocin and lasted for 12 weeks. The diabetic rats received Vilda (6 mg/kg/day, orally) for one month before I/R. Myocardial regional ischemia was induced through the ligation of left coronary artery, and IPostC was applied immediately at the onset of reperfusion. The infarct size was assessed by a computerised planimetry and left ventricles samples were harvested for cardiac mitochondrial function studies (ROS production, membrane potential and staining) and western blotting was used for determination of autophagy markers. Results: None of Vilda or IPostC but combination of them could significantly reduce the infarct size of diabetic hearts, comparing to control (P<0.001). IPostC could not significantly affect p62 expression level in diabetic hearts, but pre-treatment with Vilda alone (p<0.05) and in combination with IPostC (p<0.01) more significantly decreased p62 expression in comparison with corresponding control group. The expression of LC3B-II and LC3BII/LC3BI as well as mitochondrial ROS production were decreased significantly in treatment groups (p<0.001). Mitochondrial membrane depolarization was significantly higher and mitochondrial density was lower in untreated diabetic I/R hearts than treated groups (p<0.001). IPostC in combination with vildagliptin prevented the mitochondrial membrane depolarization and increased the mitochondrial content more potent than IPostC alone in diabetic hearts. Conclusion: Combination of vildagliptin and IPostC in diabetic hearts was a well-working strategy to reduce myocardial I/R damages by restoring mitochondrial membrane potential and ROS production and modulating the autophagic activity in I/R hearts

The Role of Stearoyl-coenzyme A Desaturase 1 in Liver Development, Function, and Pathogenesis

Stearoyl-coenzyme A desaturase 1 (SCD1) is a microsomal enzyme that controls fatty acid metabolism and is highly expressed in hepatocytes. SCD1 may play a key role in liver development and hepatic lipid homeostasis through promoting monounsaturated protein acylation and converting lipotoxic saturated fatty acids into monounsaturated fatty acids. Imbalanced activity of SCD1 has been implicated in fatty liver induction, inflammation and stress. In this review, the role of SCD1 in hepatic development, function and pathogenesis is discussed. Additionally, emerging novel therapeutic agents targeting SCD1 for the treatment of liver disorders are presented