Serdar Altun

Histopathological effects, responses of oxidative stress, inflammation, apoptosis biomarkers and alteration of gene expressions related to apoptosis, oxidative stress, and reproductive system in chlorpyrifos-exposed common carp (Cyprinus carpio L.)

In this study, we aimed to identify the toxic effects of chlorpyrifos exposure on the tissues of common carp. For this purpose, we evaluated histopathological changes in the brain, gills, liver, kidney, testis, and ovaries after 21 days of chlorpyrifos exposure. Activation of 8-OHdG, cleaved caspase-3, and iNOS were assesed by immunofluorescence assay in chlorpyrifos-exposed brain and liver tissue. Additionally, we measured the expression levels of caspase-3, caspase-8, iNOS, MT1, CYP1A, and CYP3A genes in chlorpyrifos-exposed brain tissue, as well as the expression levels of FSH and LH genes in chlorpyrifos-exposed ovaries, using qRT-PCR. We observed severe histopathological lesions, including inflammation, degeneration, necrosis, and hemorrhage, in the evaluated tissues of common carp after both high and low levels of exposure to chlorpyrifos. We detected strong and diffuse signs of immunofluorescence reaction for 8-OHdG, iNOS, and cleaved caspase-3 in the chlorpyrifos-exposed brain and liver tissues. Furthermore, we found that chlorpyrifos exposure significantly upregulated the expressions of caspase-3, caspase-8, iNOS, and MT1, and also moderately upregulated CYP1A and CYP3A in the brain tissue of exposed carp. We also noted downregulation of FSH and LH gene expressions in chlorpyrifos-exposed ovary tissues. Based on our results, chlorpyrifos toxication caused crucial histopathological lesions in vital organs, induced oxidative stress, inflammation, and apoptosis in liver and brain tissues, and triggered reproductive sterility in common carp. Therefore, we can propose that chlorpyrifos toxication is highly dangerous to the health of common carp. Moreover, chlorpyrifos pollution in the water could threaten the common carp population. Use of chlorpyrifos should be restricted, and aquatic systems should be monitored for chlorpyrifos pollution.

Evaluation of 8-hydroxy-2-deoxyguanosine and NFkB activation, oxidative stress response, acetylcholinesterase activity, and histopathological changes in rainbow trout brain exposed to linuron

Linuron is a widely used herbicide to control grasses and annual broad leaf weeds. It is known that linuron has toxic effects on different organisms. However, the toxic effects of linuron on aquatic organisms, especially fish, is completely unknown. Thus, we aimed to investigate changes in 8-hydroxy-2-deoxyguanosine (8-OHdG) and nuclear factor kappa B (NFkB) activity, histopathological changes, antioxidant responses and acetylcholinesterase (AChE) activity in rainbow trout brain after exposure to linuron. Fish were exposed to 30μg/L, 120μg/L and 240μg/L concentrations of linuron for twenty-one days. Brain tissues were taken from fish for 8-OHdG and NFkB activity, histopathological examination and determination of superoxide dismutase (SOD), catalase (CAT) enzyme activity, lipid peroxidation (LPO), and reduced glutathione (GSH) levels. Our data indicated that high linuron concentrations caused a decrease in GSH levels, SOD and CAT activities in brain tissues (p<0.05). LPO levels were significantly increased by 240μg/L linuron. All concentrations caused a significant inhibition in brain AChE enzyme activity (p<0.05). Immunopositivity was detected for 8-OHdG and NFkB, and linuron exposure caused histopathological damage to the brain tissues. The results of this study can provide useful information for understanding of linuron-induced toxicity.

Acute toxication of deltamethrin results in activation of iNOS, 8-OHdG and up-regulation of caspase 3, iNOS gene expression in common carp (Cyprinus carpio L.)

Deltamethrin is a widely used synthetic pyrethroid pesticide that protects agricultural yields, including crops, fruits, and vegetables from insect-pests. It is known that deltamethrin toxication leads to metabolic disorders and has detrimental effects on the brain and liver in different organisms. However, the harmful effects of deltamethrin toxication on aquatic animals remain unclear. In the present study, we aimed to evaluate the adverse effects of deltamethrin toxication by performing a histopathological examination, an immunofluorescence assay, and a qRT-PCR on common carp. We observed that a low-dose (0.04μM) and a high-dose (0.08μM) of deltamethrin exposure caused lamellar cells hyperplasia and inflammatory cells infiltration in the gills, hyperemia, diffuse hydropic degenerations and focal necrosis in the hepatocytes, necrotic changes in the neurons, and also induced activation of inducible Nitric Oxide Synthase (iNOS) and 8-hydroxy-2-deoxyguanosine (8-OHdG) in the gills, liver, and brain depending on the exposure time (24h, 48h, 72h and 96h). In addition, deltamethrin toxication caused the up-regulation of caspase-3 and the inducible Nitric Oxide Synthase (iNOS) of the gene expression depending on the dose (0.04μM and 0.08μM) and the exposure time in the brain (p<0.05, p<0.01, p<0.001). Our results indicated that long-term deltamethrin exposure could lead to inflammation, oxidative stress, DNA damage, and apoptosis on the different organs in common carp. Thus, deltamethrin toxication is dangerous for common carp populations, and the usage of deltamethrin should be controlled and restricted in agricultural areas.

The effects of acute boric acid treatment on gill, kidney and muscle tissues in juvenile rainbow trout

Boric acid (BA) is an essential nutrient for plants and many organisms, but it has become an environmental contaminant because of widespread use. Pesticide and its compounds are a serious threat to aquatic organisms. This study was carried out to determine the histopathological effects of acute exposure to BA concentrations in rainbow trout. The fish were exposed to 102 and 103 mg/L concentrations of BA. Tissues were sampled at 6, 12, 24, 48 and 96 h. Histopathological alterations occurring in tissues were common in both doses of BA. Gill tissues showed lamellar oedema, cellulary infiltration, lamellar disorganization, degenerative changes and lamellar thickening. Kidneys had glomerular oedema and glomerulonephritis, degeneration of the tubulary epithelium, interstitial fibrosis and a hyaline cast within the tubular lumens. Muscle tissues displayed interstitial oedema and degenerative and atrophic changes to varying degrees in the myofibrils. Our study shows that BA can be toxic for rainbow trout and cause histopathological damage in fish tissue.

Ameliorative Effect of Carvacrol on Cisplatin-Induced Reproductive Damage in Male Rats.

Cisplatin (CP) treatment causes the damage in male reproductive system. Carvacrol (CARV) is an antioxidant that is naturally found in some plants. We aimed to investigate the effect of CARV on CP‐induced reproductive toxicity in male rats. Eighteen adult male Sprague–Dawley rats were used. The control group (n = 6) was treated orally with physiological saline (PS) daily for 14 days and a single intraperitoneal (IP) PS injection on day 10. The CP group (n = 6) was administered with daily oral PS for 14 days and a single IP injection of 10 mg/kg CP on day 10. The CARV + CP group (n = 6) was treated with daily 75 mg/kg oral CARV for 14 days and a single IP injection of 10 mg/kg CP on day 10. CP treatment caused the damage on some spermatological parameters (motility, live sperm rate, and abnormal sperm rate), increased the oxidative stress, and induced testicular degeneration and apoptosis. However, CARV treatment mitigates CP‐induced reproductive toxicity.

Imidacloprid exposure cause the histopathological changes, activation of TNF-α, iNOS, 8-OHdG biomarkers, and alteration of caspase 3, iNOS, CYP1A, MT1 gene expression levels in common carp (Cyprinus carpio L.)

Graphical abstract

Cypermethrin, chlorpyrifos, deltamethrin, and imidacloprid exposure up-regulates the mRNA and protein levels of bdnf and c-fos in the brain of adult zebrafish (Danio rerio)

The aim of the present study is to investigate the toxicity effects of frequently used pesticides, involving cypermethrin, deltamethrin, chlorpyrifos and imidacloprid, on the expression of bdnf and c-fos genes in zebrafish brain tissues. Therefore, brain tissues exposed to intoxication was primarily analyzed by indirect immunofluorescence assay. Afterwards, the mRNA transcription levels of BNDF and c-fos genes and the protein levels were measured by qRT-PCR and Western blotting, respectively. The data of the immunofluorescence assay revealed intensive immunopositivity for bdnf and c-fos genes in the tissues exposed to pesticide intoxication in comparison to the control group (p<0.05). Moreover, the transcription levels of BNDF and c-fos genes, and protein levels were elevated following the intoxication (p<0.05, p<0.01, and p<0.001, respectively). These results showed that the exposure to the acute cypermethrin, deltamethrin, chlorpyrifos and imidacloprid intoxication disrupted the normal neuronal activity, resulting in neurotoxic effect, also DNA-binding Increasing c-fos activation, an oncoprotein from the family of the Nuclear Proteins, is also true of the knowledge that these chemicals are oncogenic in zebrafish brain tissues. Thus, the use of these pesticides poses a potential neuronal and oncogenic risk to the non-target organisms.

Effect of Helichrysum plicatum DC. subsp. plicatum ethanol extract on gentamicin-induced nephrotoxicity in rats

The aim of this study was to evaluate the possible therapeutic or protective effects of Helichrysum plicatum DC. subsp. plicatum ethanol extract (HPE) against gentamicin-induced nephrotoxicity. Thirty-six Sprague Dawley male rats weighing between 200 and 250 g were used as live material. They were formed into six groups containing 6 rats each and were allowed to adapt to laboratory conditions for 7 d. Group I: control, 5% DMSO intraperitoneal (i.p.); Group II: HPE 100 mg/(kg·d) i.p.; Group III: HPE 200 mg/(kg·d) i.p.; Group IV: gentamicin as 80 mg/(kg·d) i.p.; Group V: gentamicin as 80 mg/(kg·d) i.p.+HPE 100 mg/(kg·d) i.p.; and Group VI: gentamicin as 80 mg/(kg·d) i.p.+HPE 200 mg/(kg·d) i.p. for 8 d. Following treatment, serum, liver, and kidney tissues were used to assess blood urea nitrogen (BUN), creatinine, enzymatic and non-enzymatic antioxidants, and lipid peroxidation. Gentamicin significantly increased serum BUN, creatinin, and liver and kidney levels of malondialdehyde (MDA). It also decreased the activity of catalase (CAT), glutathione peroxidase (GPx), and superoxide dismutase (SOD). Treatment with the HPE 100 mg/kg reversed gentamicin-induced alterations as evidenced by decreased serum BUN and creatinin, liver and kidney oxidant marker, and tubular necrosis as well as by an increase in antioxidant enzymes. It was found that HPE 200 mg/kg significantly increased liver and kidney tissue MDA levels in nephrotoxicity in rats. As a result, these findings support the proposition that HPE in 100 mg/kg dose demonstrates in the kidney and liver as free radicals and scavenger to prevent the toxic effects of gentamicin in both the biochemical and histopathology parameters.摘要目的评估蜡菊乙醇提取物(HPE)对庆大霉素诱导的 肾毒性的治疗或保护作用。方法将36 只体重200~250 g 的Sprague Dawley 雄性大 鼠分成6 组,每组6 只,适应实验室条件7 d。 每组处理方式不同,包括:组I,对照组,5% DMSO;组II,HPE 100 mg/(kg·d);组III,HPE 200 mg/(kg·d);组IV,庆大霉素80 mg/(kg·d); 组V,庆大霉素80 mg/(kg·d)+HPE 100 mg/(kg·d);组VI,庆大霉素80 mg/(kg·d)+HPE 200 mg/(kg·d)。 腹腔注射8 d 后,取血清、肝和肾组织用于评估 血液尿素氮(BUN)、肌酐、酶和非酶抗氧化剂 和脂质过氧化。结论庆大霉素能显著提升血清BUN、肌酐和肝肾阳 性以及丙二醛(MDA)水平,同时降低过氧化 氢酶(CAT),谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)的活性。用100 mg/kg HPE 的治疗能逆转庆大霉素诱导的改变。因此, 100 mg/kg HPE 在肾脏和肝脏中可作为自由基和 清除剂,具有缓解庆大霉素在生物化学和组织病 理学上毒性的作用。

The effect of diabetes on ovaries in a rat model: the role of interleukin-33 and apoptosis

Abstract Interleukin-33 (IL-33) is a novel cytokine involved in diabetes mellitus (DM) but its role in diabetic ovarian injury is unknown. As IL-33 is modulated by apoptosis, we aimed at investigating the effect of diabetes on ovaries in terms of evaluating apoptosis and IL-33 in a rat model. In this prospective experimental study, 16 female, nonpregnant Sprague–Dawley albino rats (12 weeks, 220–240 g) were randomly divided into two groups. Group 1 included eight healthy nondiabetic rats as controls and group 2 included eight rats in which diabetes was induced by intraperitoneal (i.p) injection of streptozotocin (STZ). After overt DM occurred (blood glucose >400 mgr/dl), all animals were euthanized and blood samples were collected by cardiac puncture for biochemical analysis. Bilateral oophorectomy was performed for histopathological examination. Serum levels of IL-33 and ovarian IL-33 and caspase-3 immunoexpressions were assessed. Immunoexpressions of caspase-3 and IL-33 were significantly higher in ovarian stromal cells of the diabetic rats compared to the controls. Also, in diabetic group, serum IL33 levels were significantly higher than the control group. In conclusion, increased IL-33 was observed both in serum and ovaries of STZ-induced diabetic rats as well as increased apoptosis in these diabetic rats. IL-33 may contribute to the apoptosis in diabetic ovarian injury.