Boric acid is known to regulate the proliferation of cancer cells. Prostate cancer is among the types of cancer with high mortality in men. There are a few numbers of studies investigating the ...effects of boric acid on prostate cancer cells. The objective of the present study was to assess the effects of boric acid at concentrations higher than that can be achieved in blood by dietary intake on DU-145 human prostate cancer cells for 24 h. Firstly, we determined the cytotoxic activity of boric acid (0 to 12.5 mM) on DU-145 human prostate cancer cells by using 3-(4, 5-dimethylthiazol, 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and defined the IC50 concentration of boric acid. Then, by employing the doses found in MTT, the levels of antioxidant-oxidant molecules and apoptotic proteins were measured and morphological changes were evaluated. We have concluded that boric acid caused oxidative stress, inhibition of cell growth, apoptosis, and morphological alterations in a concentration-dependent manner in DU-145 cells. Furthermore, treatments with increasing boric acid concentrations decreased the antioxidant levels in cells. We actually revealed that boric acid, known as an antioxidant, may prevent cell proliferation by acting as an oxidant in certain doses. Although the high IC50 concentration of boric acid is perceived to be negative, we think it provides important background for subsequent studies.
Prostate cancer is the main cause of cancer-related mortality in men around the world and an important health problem. DU-145 human prostate cancer cells provide an opportunity to investigate ...prostate cancer. Betaine has a number of anticancer effects, such as inactivation of carcinogens, inhibition of cancer cell proliferation, angiogenesis, and metastasis. However, there is no study investigating the effects of betaine on DU-145 cells. The aim of this study was to evaluate the effects of different concentrations of betaine on the oxidative stress, apoptosis, and inflammation on DU-145 cells. Firstly, we proved the cytotoxic activity of betaine (0 to 150 mg/ml) on DU-145 cells by using 3-(4, 5-dimethylthiazol, 2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) and defined the optimal concentration of betaine. Then, by employing the doses found in MTT, the levels of antioxidant (GSH, SOD, CAT, and TAS) and oxidant (MDA and TOS) molecules, pro-inflammatory cytokines (TNF-a and IL-6), apoptotic proteins (CYCS and CASP3), and DNA fragmentation were measured. Morphological changes and apoptosis were evaluated using H&E technique, Bax and Bcl-2 immunohistochemistry. Results suggested that betaine caused oxidative stress, inflammation, inhibition of cell growth, apoptosis, and morphological alterations in DU-145 cells dose-dependently. Furthermore, treatments with increasing betaine concentrations decreased the antioxidant levels in cells. We actually revealed that betaine, known as an antioxidant, may prevent cell proliferation by acting as an oxidant in certain doses. In conclusion, betaine may act as a biological response modifier in prostate cancer treatment in a dose-dependent manner.
Diabetes is a common metabolic disease which damages many organs including the liver and causes endoplasmic reticulum (ER) stress, which originates from non-folded proteins. Sonic hedgehog (Shh) ...pathway plays a role in liver regeneration and repair. To our knowledge, there is no study showing the relation between ER stress and Shh pathway in the liver in diabetes. Thus, the aim of this study was to investigate the interaction between ER stress and Shh pathway in the liver of diabetic mice.
Six groups of male mice were formed as control, diabetes (streptozotocine-treated), Shh activator (SAG-treated), Shh inhibitor (SANT1-treated), diabetes + SAG and diabetes + SANT1. At the end of the experiment, mice were weighed, anaesthetized and euthanized. Blood samples were collected, livers were excised and weighed. Thereafter, blood glucose, serum ALT and AST levels, TOS and TAC levels in liver tissue were measured. ER stress marker (GRP78) and Shh pathway molecules (Gli1 and Smo) were evaluated by immunohistochemistry, H-score and western blot analyses. Besides, histopathological examination was performed.
Results showed that GRP78, Gli1 and Smo were increased in liver due to Type 1 diabetes. The SAG agent decreased GRP78 and increased Gli1 and Smo, leading to liver repair, while the inhibitor SANT1 increased GRP78 and decreased Gli1and Smo, causing progression of the liver stress induced by diabetes.
In conclusion, the Shh pathway is related to ER stress and may provide a new strategy for its treatment, especially liver stress induced by diabetes.
High blood levels of β-carotene and increased intake in the diets are inversely proportional to incidence of many cancer types. Antioxidant activity of β-carotene was proposed to be related with its ...antitumor effect. Despite this plant derivative substance being sought in many cancer types, the effectiveness of β-carotene against malignant mesothelioma remained unclear. Therefore, the present study aims to explore the impact of β-carotene on cell viability, apoptosis, and oxidative stress in mesothelioma cells. Human mesothelioma cell SPC212 were treated with β-carotene (3.125–200 μM) for 24, 48, 72, and 96 h. Cytotoxicity was measured with the MTT assay (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide). Annexin-V/propidium iodide (PI) and caspase 3/7 biomarkers were used to identify apoptotic cells. Finally, the oxidative stress was evaluated with flow cytometry. The results of the measurements indicated a significant decline in viable mesothelioma cancer cell numbers upon β-carotene treatment in time- and concentration-dependent manner when compared to control cells. Furthermore, β-carotene treatment led to apoptosis induction according to both annexin V/PI and caspase 3/7 assays. Furthermore, β-carotene increased oxidative stress in SPC212 cells. These results show how β-carotene affects proliferative, apoptotic, and oxidative properties in SPC212 malignant pleural mesothelioma cells and provide useful insights into future studies.
Cyclophosphamide (CP) is a common anticancer drug, but its use in cancer treatment is limited due to its severe toxicities induced mainly by oxidative stress in normal cells. Reactive oxygen species ...(ROS) lead to multiple organ injuries, including the kidneys. Selenium (Se) is a nutritionally essential trace element with antioxidant properties. In the present study, the possible protective effect of Se on CP-induced acute nephrotoxicity was investigated. Forty-two Sprague-Dawley rats were equally divided into six groups of seven rats in each. The control group received saline, and other groups were injected with CP (150 mg/kg), Se (0.5 or 1 mg/kg), or CP + Se intraperitoneally. Total antioxidant capacity (TAC), total oxidant state (TOS), oxidative stress index (OSI), creatinine, and cystatin C (Cys C) levels were measured in the sera. In addition, kidney tissues were examined histologically. In the CP alone treated rats, creatinine, Cys C, TOS, and OSI levels increased, while TAC level decreased. CP-induced histological damages were decreased by co-treatment of Se and biochemical results supported the microscopic observations. In conclusion, our study points to the therapeutic potential of Se and indicates a significant role of ROS in CP-induced kidney toxicity.
Acrylamide is a neurotoxic agent forming in foods. Thymoquinone and quercetin are plant-derived antioxidants in various foods with known benefits. C6 cells are glioblastoma cells. In this study, we ...aimed at preventing acrylamide toxicity by thymoquinone and quercetin in the C6 cell line. In our study, first, toxic doses of acrylamide, nontoxic doses of thymoquinone and quercetin in C6 cells for 24 h were determined by MTT (3-4,5-dimethylthiazol-2-yl-2,5 diphenyl tetrazolium bromide) colorimetric test. After that, caspase 3/7 and annexin V tests were performed by flow cytometry to evaluate whether the apoptosis pathway was inducted. Furthermore, autophagy and oxidative stress were assessed by flow cytometry. The amount of Nrf2 (nuclear factor erythroid 2-related factor 2) was determined by immunocytochemistry. The morphological examination was performed by microscopic analyses. As a result, 4 mM of acrylamide was determined to be used to induce toxicity in C6 cells. The nontoxic doses of thymoquinone and quercetin were respectively determined as 3.9 and 2.0 µM. Thymoquinone and quercetin not only reduced acrylamide-induced apoptosis in annexin V and caspase 3/7 assays but also morphological deformations in microscopic examinations. In autophagy, it was revealed that acrylamide-induced autophagy was decreased by quercetin and thymoquinone pretreatments. As for Nrf2 expression, it was observed that acrylamide increased Nrf2 expression, and thymoquinone and quercetin pretreatments increased it even further. In conclusion, in the study, acrylamide demonstrated a damaging effect on C6 glioblastoma cells, and thymoquinone and quercetin pretreatments exerted a protective effect against acrylamide-induced damage in C6 cells.
•Acrylamide induces dose-dependent toxic effect on BEAS-2B normal lung cells.•Apoptotic hallmarks and signs are observable after acrylamide treatment.•Nrf2 translocates to nucleus and Nrf-2 ...expression increases following acrylamide injection.•IC50 and IC75 of acrylamide for BEAS-2B cells are 2.0 and 2.75 mM, respectively.•BEAS-2B cells are very sensitive to acrylamide treatment when compared to other cell lines.
Due to the broad toxic relevance of acrylamide, many measures have been taken since the 1900s. These measures increased day by day when acrylamide was discovered in foods in 2002, and its toxic spectrum was found to be wider than expected. Therefore, in some countries, the products with higher acrylamide content were restricted. On the other hand, the effects of acrylamide on the respiratory system cells have yet to be well understood. In this study, we aimed at investigating the effect of acrylamide on lung epithelial BEAS-2B cells. Initially, the cytotoxic effect of acrylamide on BEAS-2B was determined by MTT assay. Then, cellular oxidative stress was measured. Flow cytometry analysis was conducted for Annexin-V and caspase 3/7. Furthermore, Bax, Bcl-2 and Nrf-2 proteins were evaluated by immunocytochemistry. Finally, acrylamide-induced cellular morphological changes were observed under confocal and TEM microscopes. According to MTT results, the IC50 concentration of acrylamide was 2.00 mM. After acrylamide treatment, oxidative stress increased dose-dependently. Annexin V-labelled apoptotic cells and caspase 3/7 activity were higher than untreated cells in acrylamide-treated cells. Immunocytochemical examination revealed a marked decrease in Bcl-2, an increase in Bax and Nrf-2 protein staining upon acrylamide treatment. Furthermore, in confocal and TEM microscopy, apoptotic hallmarks were pronounced. In the present study, acrylamide was suggested to display anti-proliferative activity, decrease viability, induce apoptosis and oxidative stress and cause morphological changes in BEAS-2B cells.
Acrylamide is a dangerous electrophile with the potency to react with many biological moieties including proteins, and nucleic acids as well as other macromolecules. Acrylamide was first only known a ...chemical exposed in working areas as a neurotoxicant, it was later discovered that beyond just being a neurotoxicant exposed in industrial areas, acrylamide is exposed via daily foods as well. As such, several strategies have been sought to be developed to relieve the toxic spectrum of this chemical. The utilization of a protective agent against acrylamide toxicity was one of those strategies. To date, many agents with protective potency have been investigated. Herein, we compiled these agents and their effects shown in in vitro studies. We used the search engines of Web of Knowledge and searched the keywords "acrylamide" and "protect" in the titles along with the keyword "cell" in the topics. Twenty-one directly related articles out of 35 articles were examined. Briefly, all agents used against acrylamide were reported to exhibit protective activity. In most of these reports, 5 mM concentration of acrylamide and 24-hour treatment were the employed dose and duration. Usually, the beneficial agents were pre-treated to the cells. PC12 cells were the most utilized cell line, and the mitogen-activated protein kinase (MAPK) and nuclear factor erythroid 2-related factor 2 (NRF2) pathways were the most studied pathways. This study, beside other importance, can be utilized as a guide for how the protective studies against acrylamide were done and which parameters were investigated in in vitro acrylamide studies. In conclusion, taking measures is of utmost importance to prevent or alleviate the toxicity of acrylamide, to which we are daily exposed even in our homes. Therefore, future studies should persist in focusing on mitigating acrylamide toxicity.
This study aims to investigate protective effects of boron against cyclophosphamide-induced bladder toxicity that produces oxidative stress and leads to apoptosis of the cells. In total, 24 rats were ...divided into 4 equal groups. The control group received saline. The 2nd experimental group received 200 mg kg of cyclophosphamide i.p. on the 4th day while the 3rd group was given only boron (200 mg kg, i.p.) for 6 days. In the 4th group, boron was given for 6 days and cyclophosphamide (200 mg kg, i.p.) was administrated on the 4th day. Twenty-four hours after the last boron or cyclophosphamide administration, rats were sacrificed under anesthesia. Bladder tissues of rats were taken for histological and immunohistochemical (apoptotic markers such as caspase-3, bcl-2, and bax) and blood was taken for the biochemical (serum total thiol, serum natural thiol, serum thiol-disulfide) analysis. Transient epithelial thinning, edema, marked inflammatory reaction, and bleeding were observed in bladders of the group that received cyclophosphamide. Also, the activity of bax and caspase-3-positive cells increased while the number of bcl-2-positive cells decreased. In the same group, serum natural thiol and total thiol levels decreased while serum disulfide levels increased, which indicates oxidative stress. On the other hand, in the boron+cyclophosphamide group pretreatment with boron protected, the bladder tissue and the number of bcl-2-positive cells increased, and bax and caspase-3-positive cells decreased, showing antiapoptotic effects of boron against cyclophosphamide-induced toxicity. In parallel with the findings of this group, native thiol and total thiol levels increased and serum disulfide levels decreased pointing out to a decreased oxidative stress. Our results indicate that boron pretreatment significantly protects rat bladder against cyclophosphamide-induced bladder damage due to its antiapoptotic and antioxidant properties.
This study aims to investigate the protective effects of silymarin (Sm) in thioacetamide (TAA)-related liver damage. What makes this study special is that it attempts to determine the expression of ...changes in the liver at the level of gene expression. Routine liver damage markers were compared with changes in the levels of microRNA (miRNA) known as new biomarkers. With this in mind, we divided the rats into four groups including control, TAA, Sm + TAA (50 + 50 mg/kg), and Sm + TAA (100 + 50 mg/kg). Blood and tissue samples belonging to the rats were collected in consideration of morphological, immunohistochemistry, miRNAs levels, and biochemical evaluations. Our study results showed that miR-122, miR-192, and miR-194 levels had decreased in the experimental groups given TAA, whereas miR-122, miR-192, and miR-194 levels had increased in the doses of Sm + TAA-given group. Therefore, Sm treatment undertaken before exposure to the toxin successfully altered its effects upon the study animals.
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