Stereological Study of the Protective Role of Curcumin on Histological Changes of Seminal Vesicle in Mice Following Treatment with Sodium Arsenite

The catabolism of heme is carried out by members of the heme oxygenase (HO) family. The products of heme catabolism by HO-1 are ferrous iron, biliverdin (subsequently converted to bilirubin), and carbon monoxide. In addition to its function in the recycling of hemoglobin iron, this microsomal enzyme has been shown to protect cells in various stress models. Implicit in the reports of HO-1 cytoprotection to date are its effects on the cellular handling of heme/iron. However, the limited amount of uncommitted heme in non-erythroid cells brings to question the source of substrate for this enzyme in non-hemolytic circumstances. In the present study, HO-1 was induced by either sodium arsenite (reactive oxygen species producer) or hemin or overexpressed in the murine macrophage-like cell line, RAW 264.7. Both of the inducers elicited an increase in active HO-1; however, only hemin exposure caused an increase in the synthesis rate of the iron storage protein, ferritin. This effect of hemin was the direct result of the liberation of iron from heme by HO. Cells stably overexpressing HO-1, although protected from oxidative stress, did not display elevated basal ferritin synthesis. However, these cells did exhibit an increase in ferritin synthesis, compared with untransfected controls, in response to hemin treatment, suggesting that heme levels, and not HO-1, limit cellular heme catabolism. Our results suggest that the protection of cells from oxidative insult afforded by HO-1 is not due to the catabolism of significant amounts of cellular heme as thought previously.

Arsenic exposure is well documented to cause serious health hazards, such as cardiovascular abnormalities, neurotoxicity and nephrotoxicity. In the present study, we intended to explore the role of bosentan, an endothelial receptor antagonist, against sodium arsenite-induced nephrotoxicity and hepatotoxicity in rats. Sodium arsenite (5 mg/kg, oral) was administered for 4 weeks to induce renal dysfunction in rats. Sodium arsenite intoxicated rats were treated with bosentan (50 and 100 mg/kg, oral) for 4 weeks. Arsenic led renal damage was demonstrated by significant increase in serum creatinine, urea, uric acid, potassium, fractional excretion of sodium, microproteinuria and decreased creatinine clearance in rats. Sodium arsenite resulted in marked oxidative stress in rat kidneys as indicated by profound increase in lipid peroxides, and superoxide anion generation alongwith decrease in reduced glutathione levels. Hydroxyproline assay highlighted arsenic-induced renal fibrosis in rats. Hematoxylin-eosin staining indicated glomerular and tubular changes in rat kidneys. Picrosirius red staining highlighted collagen deposition in renal tissues of arsenic treated rats. Immunohistological results demonstrated the reduction of renal eNOS expression in arsenic treated rats. Notably, treatment with bosentan attenuated arsenic-induced renal damage and resisted arsenic-led reduction in renal eNOS expression. In addition, sodium arsenite-induced alteration in hepatic parameters (serum aspartate aminotransferase, alanine transferase, alkaline phosphatase, bilirubin), oxidative stress and histological changes were abrogated by bosentan treatment in rats. Hence, we conclude that bosentan treatment attenuated sodium arsenite-induced oxidative stress, fibrosis and reduction in renal eNOS expression in rat kidneys. Moreover, bosentan abrogated arsenic led hepatic changes in rats.

Epicatechin ameliorates glucose intolerance and hepatotoxicity in sodium arsenite-treated mice

We recently reported that arsenic caused insulin resistance in differentiated human neuroblastoma SH-SY5Y cells. Herein, we further investigated the effects of sodium arsenite on IGF-1 signaling, which shares downstream signaling with insulin. A time-course experiment revealed that sodium arsenite began to decrease IGF-1-stimulated Akt phosphorylation on Day 3 after treatment, indicating that prolonged sodium arsenite treatment disrupted the neuronal IGF-1 response. Additionally, sodium arsenite decreased IGF-1-stimulated tyrosine phosphorylation of the IGF-1 receptor β (IGF-1Rβ) and its downstream target, insulin receptor substrate 1 (IRS1). These results suggested that sodium arsenite impaired the intrinsic tyrosine kinase activity of IGF-1Rβ, ultimately resulting in a reduction in tyrosine-phosphorylated IRS1. Sodium arsenite also reduced IGF-1 stimulated tyrosine phosphorylation of insulin receptor β (IRβ), indicating the potential inhibition of IGF-1R/IR crosstalk by sodium arsenite. Interestingly, sodium arsenite also induced neurite shortening at the same concentrations that caused IGF-1 signaling impairment. A 24-h IGF-1 treatment partially rescued neurite shortening caused by sodium arsenite. Moreover, the reduction in Akt phosphorylation by sodium arsenite was attenuated by IGF-1. Inhibition of PI3K/Akt by LY294002 diminished the protective effects of IGF-1 against sodium arsenite-induced neurite retraction. Together, our findings suggested that sodium arsenite-impaired IGF-1 signaling, leading to neurite shortening through IGF-1/PI3K/Akt.

Arsenic is a universal component and a notable natural poison. Its exposure to people occurs primarily through natural, therapeutic, and occupational sources. This investigation intended to discover the defensive impact of metformin (100 and 150 mg/kg body weight) against sodium arsenite (SA)-induced cardiotoxicity in experimental animals. The study was conducted on Sprague Dawley rats (n = 50), which were separated into five different groups as follows: control, met-150, SA, SA + met-100, and SA + met-150. The results demonstrated that SA caused a significant increase in the level of malondialdehyde and also reduced activities of antioxidative enzyme. SA similarly increased inflammatory reactions by increasing the level of interleukin-6, tumor necrosis factor-α, and interleukin-1β. In addition, SA provoked the apoptosis by expanding the p53 and Bax levels, terminal deoxynucleotidyl transferase dUTP nick end labeling, and expression of caspase-3. SA altered the histological integrity of cardiac tissue and 8-hydroxy-2'-deoxyguanosine expression. In conclusion, metformin significantly reduced oxidative stress, inflammatory reaction, and apoptotic pathway. The present investigation showed that metformin has a cardioprotective impact because of its protective role against oxidation, inflammation, and apoptosis.

Introduction: To study the protective effect of extra virgin olive oil (EVOO) on histopathological changes induced by arsenic in the kidneys of albino rats. Randomized control trial. November 1, 2017–November 30, 2017 at National Institute of Health. Material and Methods: Forty-five male adult albino rats were placed in three cages having 15 rats each. Distilled water was given to the rats of control Group I for 30 days. The dose of sodium arsenite given to rats was 40 mg per kg per day dissolved in drinking water for 30 days. Olive oil of 0.2 ml per day was only given to Group III rats for 30 days along with sodium arsenite. In dissection was done after 30 days and their kidneys were approached and dissected out for histological changes. Results: EVOO has ameliorated the microscopic quantitative and qualitative histological changes induced by arsenic on both kidneys of albino rats. Olive oil had significantly prevented the increase in the diameter of the proximal convoluted tubule (PCT). Discussion and Conclusion: The present study demonstrates that EVOO prevents the quantitative and qualitative histological changes caused by arsenic on kidneys which include the diameter of PCT and distal convoluted tubule, the diameter of glomeruli, width of bowman’s space, and loss of brush border.

Arsenite is a major environmental chemical and a known reproductive toxicant via the depression of spermatogenesis and androgenesis in males. The possibility of sodium arsenite reproductive toxicity been caused by autooxidation was investigated in this study taking advantage of the anti-oxidant properties of ginger and its androgenic activities. The effect of exposure to sodium arsenite (10 mg/kg BW/day) by gavage via oral cannula without or with aqueous ginger extract (500mg/kg BW/day) co-treatments for 30 days was evaluated in adult male rats. The weight of the reproductive organs, sperm count, motility, and morphology were evaluated. Plasma FSH, LH and testosterone levels were assayed. Lipid peroxidation (indexed by MDA) and antioxidants enzymes likes GSH, SOD, CAT were assessed. Sodium arsenite treatment decreased the reproductive organs weight: testis, epididymis, prostate and seminal vesicle; sperm functions: count, motility and normal morphology; plasma hormones level: FSH, LH and testosterone. There was a decrease in the activities of GSH, SOD and CAT as well as an increase in MDA concentration. Co-administration of aqueous ginger extract with arsenite was found to protect against adverse change in the reproductive organ weight, attenuate the decrease in sperm functions, enhance plasma reproductive hormones level along with increased antioxidants activities and reduced peroxidation. This study showed that sodium arsenite apart from being a hormonal disrupter also causes oxidative stress which contributed to the reproductive damage in the male rats. The protective effects of ginger on reproductive toxicity and oxidative stress as evidenced by the clear restoration of sperm functions, testicular steroidogenesis and reproductive organo-somatic indices could be attributed to its antioxidants and androgenic properties. Key Words : Arsenite, Ginger, Antioxidants and Sperm.

Treatment of melanoma cells by sodium arsenite or statins (simvastatin and lovastatin) dramatically modified activities of the main cell signaling pathways resulting in the induction of heme oxygenase-1 (HO-1) and in a downregulation of cyclooxygenase-2 (COX-2) protein levels. Through heme degradation and the production of carbon monoxide and biliverdin, HO-1 plays a protective role in different scenario of oxidative stress followed by mitochondrial apoptosis. Both sodium arsenite and statins could be efficient inducers of apoptosis in some melanoma cell lines, but often exhibited only modest proapoptotic activity in others, due to numerous protective mechanisms. We demonstrated in the present study that treatment by sodium arsenite or statins with an additional inhibition of HO-1 expression (or activation) caused a substantial upregulation of apoptosis in melanoma cells. Sodium arsenite- or statin-induced apoptosis was independent of BRAF status (wild type versus V600E) in melanoma lines. Monotreatment required high doses of statins (20-40 μM) for effective induction of apoptosis. As an alternative approach, pretreatment of melanoma cells with statin at decreased doses (5-20 μM) dramatically enhanced TRAIL-induced apoptosis, due to suppression of the NF-κB and STAT3-transcriptional targets (including COX-2) and downregulation of cFLIP-L (a caspase-8 inhibitor) protein levels. Furthermore, combined treatment with sodium arsenite and TRAIL or simvastatin and TRAIL efficiently induced apoptotic commitment in human neuroblastoma cells. In summary, our findings on enhancing effects of combined treatment of cancer cells using statin and TRAIL provide the rationale for further preclinical evaluation.

Regarding the strong antioxidant properties of Rosa damascene extract, this study aimed to investigate the protective role of Rosa damascene Miller hydroalcoholic petal extract on oxidative stress parameters and testis tissue in rats treated with sodium arsenite. To this end, 30 male rats were divided into five groups, including control, positive control (treated with arsenite), and three groups of patients affected by sodium arsenite with 150 mg/kg, 300 mg/kg, and 450 mg/kg Rosa damascene extract for 34 days by gavage. The animals were then anesthetized, and the blood samples were collected from the heart. The left testis was removed for histopathological studies. The findings revealed that Sodium arsenite in the positive group caused a significant reduction in TAC, testosterone, and serum Luteinizing hormone (LH) and a significant increase in serum Malondialdehyde. In addition, there was no statistically significant difference among the groups regarding the amount of Follicle-stimulating hormone (FSH). Moreover, the consumption of Rosa damascene extract with sodium arsenite caused a significant increase in testosterone, LH, and FSH compared to the positive control group. Histopathological results showed that in the experimental group receiving a dosage of 300 mg/kg b.w and the control group, the number of sperm tubes increased, and the germinal epithelium’s thickness was appropriate. Daily treatment with Rosa damascene extract with a dosage of 300 mg/kg b.w for 34 days could improve the changes caused by sodium arsenite and reduce Malondialdehyde levels. Thus, it seems that Rosa damascene hydroalcoholic extract can effectively improve the male reproductive system’s function.

This study examines histometrical changes induced by sodium arsenite (SA), as an environmental pollutant, and investigates the protective effect of α-tocopherol on ovaries of SA-treated rats during the prenatal stage until sexual maturity. Rats were classified into groups: control, SA (8 ppm/day), α-tocopherol (100 ppm/day), and SA+α-tocopherol. Treatment was performed from pregnancy until maturation when the rats and ovaries were weighed. The Cavalieri method was used to estimate volume of the ovaries, cortex, medulla, and corpus luteum. The mean diameter of oocytes, granulosa cells, and nuclei were measured and volume was estimated using the Nucleator method. The number of oocytes and thickness of the zona pellucida (ZP) were determined using an optical dissector and orthogonal intercept method, respectively. SA reduced the body and ovary weight, the number of secondary, antral and Graafian oocytes, volume of the ovaries, cortex, medulla and corpus luteum, mean diameter and volume of oocytes in primordial and primary follicles, mean diameter and volume of oocyte nuclei in all types of follicles, and mean thickness of the ZP in secondary and antral follicles. Also, the mean diameter and volume of granulosa cells and their nuclei in antral and Graafian follicles decreased significantly. Vacuolization and vascular congestion in the corpus luteum and an increase in the number of atretic oocytes were seen in the SA group. Most of these parameters were unchanged from the control level in the SA+α-tocopherol group. It was concluded that α-tocopherol supplementation reduced the toxic effects of SA exposure on ovarian tissue in rats.

Role of Nicotinic and Estrogen Signaling during Experimental Acute and Chronic Bladder Inflammation

Induction of heat shock proteins (HSPs) is thought to play a protective role in ischaemic acute renal failure (ARF). However the role of HSPs in nephrotoxic ARF is not well explored. The aim of this study was to clarify the effects of the induction of HSP70s on cisplatin (CDDP) (6 mg/kg i.v.)-induced ARF in rats. Uranyl acetate (UA) or sodium arsenite (SA) were administered i.v. 14 days or 1 day respectively before CDDP injection to induce HSPs. Serum creatinine (SCr), tubular damage score and the numbers of apoptotic (TUNEL-positive) cells were examined 5 days after CDDP injection. The expression of HSP72, B-cell lymphoma gene product-2 (Bcl-2) and Bax were evaluated by Western blot analysis. We also investigated the effect of co-administration of chelerythrine chloride (Chel), which inhibits the induction of HSPs, with SA on the expression of HSP72 and nephrotoxicity. Pretreatment with UA or SA significantly induced renal HSP72 expression. Both UA and SA attenuated the CDDP-induced increase in SCr and tubular damage scores. Co-administration of Chel with SA abolished the SA-induced increment of HSP72 and the beneficial effects of SA. The protective effects of the induction of HSP72 were associated with an increased renal Bcl-2/Bax ratio and the reduction of TUNEL-positive cells in the outer stripe of outer medulla. Our findings suggest that HSP72 attenuates CDDP-induced nephrotoxicity. The protective effects of HSP72 are associated with an increased Bcl-2/Bax ratio and less apoptosis.

Exposure to higher levels of arsenic is a serious threat affecting human health worldwide. We investigated the protective role of betaine (N,N,N-trimethylglycine) against sodium arsenite-induced renal dysfunction in rats. Sodium arsenite (5 mg/kg, oral) was given to rats for 4 weeks to induce nephrotoxicity. Betaine (125 and 250 mg/kg, oral) was administered in rats for 4 weeks along with sodium-arsenite feeding. Arsenic-induced renal dysfunction was demonstrated by measuring serum creatinine, creatinine clearance, urea, uric acid, potassium, fractional excretion of sodium, and microproteinuria. Oxidative stress in rat kidneys was determined by assaying thiobarbituric acid reactive substances, superoxide anion generation, and reduced glutathione levels. Furthermore, hydroxyproline assay was done to assess renal fibrosis in arsenic intoxicated rats. Hematoxylin-eosin and picrosirius red staining revealed pathological alterations in rat kidneys. Renal endothelial nitric oxide synthase (eNOS) expression was determined by immuno-histochemistry. Concurrent administration of betaine abrogated arsenic-induced renal biochemical and histological changes in rats. Betaine treatment significantly attenuated arsenic-induced decrease in renal eNOS expression. In conclusion, betaine is protective against sodium arsenite-induced renal dysfunction, which may be attributed to its anti-oxidant activity and modulation of renal eNOS expression in rat kidneys.

In most tropical developing countries, one of the problems facing aquaculture industry is the pollution of ponds and rivers with pesticides. Chemicals such as diazinon, an organophosphate pesticide, originating from agricultural activity enter the aquatic environment through atmospheric deposition, surface run-off or leaching. Pollutants enter the food chain through accumulation in soft bottom sediment and aquatic organisms. However, information on how these pesticides affect inhabiting organisms is often not available. In a triplicate experimental set-up, seventy-two (72) apparently healthy catfish comprising adult and juvenile of both sexes were therefore exposed to a previously determined no effect concentration (0.405 ppm) of diazinon. Another set of fish was exposed to 0.0625 μg sodium arsenite, a known clastogen, which was used as the positive control, while another set of catfish exposed to the culture water alone was the negative control. Adults and juveniles were exposed separately to avoid cannibalism. After 48 hours of exposure, micronuclei induction was determined in subsets of experimental groups, while exposure continued for 28 days. Catfish organs were harvested on days 21 and 28 to determine the effect of long-term exposure to diazinon on histology. Water quality was also monitored before and during exposure in the experimental groups. The result established a significantly high mean micronucleated polychromatic erythrocytes (15.00) in catfish exposed to diazinon suggesting genetic damage (normal is ≤4). The MPE in sodium arsenite exposed fish was 28, while that of the control group was below 4. Effect of sex and age on micronuclei induction was not significant. Histological alteration observed in the ovary and testis was distorted matured cells and extensive testicular degeneration, respectively. The results show that diazinon has clastogenic effect, and may have endocrine disrupting properties because of the histological changes induced in the ovaries and testis.

Deposition of arsenic in mice through groundwater is well documented but little is known about the histological changes of organs by the metalloid. Present study was designed to evaluate arsenic-induced histological alterations in kidney, liver, thoracic artery and brain of mice which are not well documented yet. Swiss albino male mice were divided into 2 groups and treated as follows: Group 1: control, 2: arsenic (sodium arsenite at 10 mg/kg b.w. orally for 8 wks). Group 2 showed marked degenerative changes in kidney, liver, thoracic artery, and brain whereas Group 1 did not reveal any abnormalities on histopathology. We therefore concluded that arsenic induces histological alterations in the tested organs.