Antigenotoxic properties of Paliurus spina-christi Mill fruits and their active compounds.

BACKGROUND: Paliurus spina-christi Mill. (PS) fruits are widely used for different medical purposes in Turkey. Like in many medicinal herbs the studies concerning their activity, the activities of PS are also not well clarified. The aim of this study is to evaluate the antigenotoxicity of the compounds isolated and identified from the extracts of PS fruits. METHODS: The active compounds were separated, isolated, and determined by chromatographic methods and their structural elucidation was performed by Nuclear Magnetic Resonance (NMR) methods. The compounds were obtained from either ethyl acetate (EA) or n-butanol extracts. The cytotoxicities of the compounds using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and the antigenotoxic activities of the compounds using the alkaline single cell gel electrophoresis techniques (comet assay) were evaluated in Chinese hamster lung fibroblast (V79) cell lines. RESULTS: The isolated major compounds were identified as (+/-) catechins and gallocatechin from EA fraction and rutin from n-butanol fraction of PS fruits. Their chemical structures were identified by 1H-NMR, 13C-NMR, HMBC, and HMQC techniques. Half-maximal inhibitory concentration of catechins, gallocatechin, and rutin were found to be 734 µg/mL, 220 µg/mL, and 1004 µg/mL, respectively. The methanolic extract of PS (1-100 µg/mL) alone did not induce DNA single-strand breaks while catechins (1-100 µg/mL), gallocatechin (1-50 µg/mL), and rutin (1-50 µg/mL) significantly reduced H2O2-induced DNA damage. CONCLUSION: It has been suggested that PS fruits and their compounds catechins, gallocatechin and rutin may have beneficial effects in oxidative DNA damage. It seems that PS fruits may be used in protection of the disorders related to DNA damage.

Surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin in urine.

We present a surface-enhanced Raman probe (SERS) platform for the determination of a prohibited substance, recombinant erythropoietin (rEPO), in urine matrix, using nanoparticles as substrate. Rod-shaped gold nanoparticles (GNR) were modified with a Raman label and an antibody as SERS probe. We developed two SERS-based immunoassays for detection and quantification of rEPO in urine. In the first assay, rEPO was determined by a sandwich assay with gold surfaces and GNR. In the second assay, rEPO was extracted by using core shell-structured magnetic iron oxide gold nanoparticles, and again sandwich assay was performed by using GNR. We also demonstrated the ability of the proposed method to discriminate rEPO and urinary erythropoietin (uEPO). A good linear correlation was obtained between logarithms of rEPO concentrations in urine and Raman intensities within the range of 10-1-103 pg mL-1 rEPO concentrations. Detection limits which are smaller than 0.1 pg mL-1 levels were achieved owing to the high extractive performance of the nanoextraction techniques. Graphical Abstract Schematic represantation of surface-enhanced Raman probe for rapid nanoextraction and detection of erythropoietin.

Anticarcinogenic effects of the ethanolic extract of Salix aegyptiaca in colon cancer cells: involvement of Akt/PKB and MAPK pathways.

The bark from Salix species of plants has been traditionally consumed for its antiinflammatory properties. Because inflammation frequently accompanies the progress of colorectal cancer (CRC), we have evaluated the anticancer properties of the ethanolic extract from the bark (EEB) of S. aegyptiaca, a Salix species endogenous to the Middle East, using HCT-116 and HT29 CRC cell lines. Fresh bark from S. aegyptiaca was extracted with ethanol, fractionated by solvent-solvent partitioning and the fractions were analyzed by tandem mass spectrometry. Catechin, catechol, and salicin were the most abundant constituents of the extract. Interestingly, EEB showed the highest anticancer effect in the colon cancer cells followed by its fractions in ethyl acetate and water, with catechin, catechol, and salicin showing the least efficacy. EEB could strongly reduce the proliferation of the cancer cells, but not of CCD-18Co, normal colon fibroblast cell line. Accompanying this was cell cycle arrest at G1/S independent of DNA damage in the cancer cells, induction of apoptosis through a p53 dependent pathway and an inhibition of PI3K/Akt and MAP Kinase pathways at levels comparable to known commercial inhibitors. We propose that the combination of the polyphenols and flavonoids in EEB contributes toward its potent anticarcinogenic effects. [Supplementary materials are available for this article. Go to the publisher's online edition of Nutrition and Cancer for the following free supplemental resource(s): Supplementary Figure 1 and Supplementary Figure 2.].

Antioxidant and antigenotoxic effects of lycopene in obstructive jaundice.

BACKGROUND: Obstructive jaundice, a frequently observed condition caused by obstruction of the common bile duct or its flow and seen in many clinical situations, may end up with serious complications like sepsis, immune depression, coagulopathy, wound breakdown, gastrointestinal hemorrhage, and hepatic and renal failures. Intrahepatic accumulation of reactive oxygen species is thought to be an important cause for the possible mechanisms of the pathogenesis of cholestatic tissue injury from jaundice. Carotenoids have been well described that are able to scavenge reactive oxygen species. Lycopene, a carotenoid present in tomatoes, tomato products, and several fruits and vegetables, have been suggested to have antioxidant activity, so may play a role in certain diseases related to the oxidative stress. The aim of the present study was to determine the effects of lycopene on oxidative stress and DNA damage induced by experimental biliary obstruction in Wistar albino rats. MATERIALS AND METHODS: Daily doses of 100 mg/kg lycopene were given to the bile duct-ligation (BDL) rats orally for 14 days. DNA damage was evaluated by an alkaline comet assay. The levels of aspartate transferase, amino alanine transferase, gamma glutamyl transferase, alkaline phosphatase, and direct bilirubin were analyzed in plasma for the determination of liver functions. The levels of malondialdehyde, reduced glutathione, nitric oxide, catalase, superoxide dismutase, and glutathione S transferase were determined in the liver and kidney tissues. Pro-inflammatory cytokine tumor necrosis factor-alpha level was determined in the liver tissues. Histologic examinations of the liver and kidney tissues were also performed. RESULTS: According to this study, lycopene significantly recovered the parameters of liver functions in plasma, reduced malondialdehyde and nitric oxide levels, enhanced reduced glutathione levels, as well as enhancing all antioxidant enzyme activity in all tissues obtained from the BDL group. Moreover, the parameters of DNA damage in the liver and kidney tissue cells, whole blood cells, and lymphocytes were significantly lower in the lycopene-treated BDL group, compared with the BDL group. CONCLUSIONS: Lycopene significantly reduced the DNA damage, and markedly recovered the liver and kidney tissue injuries seen in rats with obstructive jaundice.

Preventive role of Pycnogenol® against the hyperglycemia-induced oxidative stress and DNA damage in diabetic rats.

Diabetes mellitus, a complex progressive metabolic disorder, leads to some oxidative stress related complications. Pycnogenol® (PYC), a plant extract obtained from Pinus pinaster, has been suggested to be effective in many diseases including diabetes, cancer, inflammatory and immune system disorders. The mechanisms underlying the effects of PYC in diabetes need to be elucidated. The aim of this study was to determine the effects of PYC treatment (50 mg/kg/day, orally, for 28 days) on the DNA damage and biochemical changes in the blood, liver, and kidney tissues of experimental diabetic rats. Changes in the activities of catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione-S-transferase enzymes, and the levels of 8-hydroxy-2'-deoxyguanosine, total glutathione, malondialdehyde, insulin, total bilirubin, alanine aminotransferase, aspartate aminotransferase, gamma-glutamyl transferase, high density lipoprotein, low density lipoprotein, total cholesterol, and triglyceride were evaluated. DNA damage was also determined in the whole blood cells and the liver and renal tissue cells using the alkaline comet assay. PYC treatment significantly ameliorated the oxidative stress, lipid profile, and liver function parameters as well as DNA damage in the hyperglycemic rats. The results show that PYC treatment might improve the hyperglycemia-induced biochemical and physiological changes in diabetes.

Effects of cinnamic acid on complications of diabetes

Background/aim: Diabetes mellitus (DM) is a major health problem worldwide. Cinnamic acid (CA) and its derivatives are synthesized in plants and increasing attention has been given to them in recent years due to the high number of beneficial health properties attributed to their consumption. The aim of this study was to investigate the effects of CA on streptozotocin-induced diabetes in Wistar albino rats. Materials and methods: DNA damage was evaluated in the blood, liver, and kidney cells of rats by the alkaline comet assay. Oxidative stress parameters such as catalase, superoxide dismutase, glutathione reductase, glutathione-S-transferase, and glutathione peroxidase activities and 8-hydroxy-2-deoxyguanosine, total glutathione, and malondialdehyde levels; biochemical parameters including insulin, total bilirubin, and BCA protein levels; hepatic enzyme levels such as alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and gamma-glutamyl transferase; and lipid profile parameters including high-density lipoprotein, low-density lipoprotein, total cholesterol, and triglyceride levels were also evaluated. Results: DM caused genotoxic damage and alterations in lipid profiles, oxidative stress parameters, and hepatic enzymes levels. CA treatment ameliorated these effects. Conclusion: It seems that CA might have a role in the prevention of the complications of diabetes.

Resveratrol Protects Sepsis-Induced Oxidative DNA Damage in Liver and Kidney of Rats.

BACKGROUND: The increases of free radicals have been proposed to be involved in the pathogenesis of sepsis, which leads to multiple-organ dysfunction syndromes. The uses of antioxidants as a complementary tool in the medical care of oxidative stress-related diseases have attracted attention of researchers. Resveratrol (RV) has suggested being antioxidant, anti-proliferative, and anti-inflammatory effects in various experimental models and clinical settings. AIMS: This study was undertaken to evaluate the protective effects of RV on oxidative DNA damage induced by sepsis in the liver and kidney tissues of Wistar albino rats. STUDY DESIGN: Animal experimentation. METHODS: Four experimental groups consisting of eight animals for each was created using a total of thirty-two male Wistar albino rats. Sham group was given 0.5 mL of saline intra-peritoneal (ip) only following laparatomy. Sepsis group was given 0.5 mL saline ip only following the induction of sepsis. RV-treated group was given a dose of 100 mg/kg ip RV in 0.5 mL saline following laparatomy. RV-treated sepsis group was given 100 mg/kg ip RV in 0.5 mL saline following the induction of sepsis. A model of sepsis was created by cecal ligation and puncture technique. In the liver and kidney tissues, oxidative stress parameters (malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPX)) and a proinflammatory cytokine (tumor necrosis factor alpha (TNF-alpha)), were evaluated spectrophotometrically and DNA damage was determined by the alkaline single cell gel electrophoresis (comet assay) technique using formamidopyrimidine DNA glycosylase protein. RESULTS: In the RV-treated sepsis group, the levels of MDA and TNF-alpha were lower and GSH levels, SOD and GPX activities were higher than in the septic rats (p<0.05). RV treatment significantly reduced the sepsis-induced oxidative DNA damage in the liver and kidney cells (p<0.05). CONCLUSION: It is suggested that RV treatment might reduce the sepsis-induced oxidative DNA damages in sepsis-related diseases; however, there is a need for more studies to clear up the protective mechanisms of RV against sepsis.

Hepatoprotective and antioxidant effects of lycopene in acute cholestasis.

BACKGROUND/AIM: Lycopene, which is suggested to be a potent antioxidant, may play a protective role in diseases related to oxidative stress. In order to understand the effects of lycopene in the pathogenesis of cholestasis, we investigated the effects of lycopene on oxidative stress parameters and DNA damage induced by experimental biliary obstruction in the liver tissues and the lymphocytes of Wistar albino rats. MATERIALS AND METHODS: The animals were randomized into 3 groups. The sham group was subjected to a sham operation, the BDL group was subjected to bile duct ligation (BDL), and the BDL+L group was subjected to BDL and treated with 10 mg/kg body weight of lycopene. After 7 days of treatment, the liver functions, oxidative stress parameters, and DNA damage were evaluated. RESULTS: The lycopene treatment significantly ameliorated the liver function parameters in BDL rats. It significantly reduced malondialdehyde and nitric oxide levels and enhanced reduced glutathione levels and catalase, superoxide dismutase, and glutathione S transferase activities in the BDL rats. The lycopene treatment also decreased DNA damage as assessed by comet assay in the lymphocytes and hepatocytes of the BDL rats. CONCLUSION: These results suggest that lycopene might have protective effects on acute cholestasis.

Assessment of the cytotoxic, genotoxic, and antigenotoxic potential of Pycnogenol® in in vitro mammalian cells.

Pycnogenol® (PYC), a standardized plant extract obtained from the bark of the French maritime pine Pinus pinaster, has been suggested to exert strong antioxidant activity and used as a phytochemical remedy for various diseases. In this study, we investigated the antioxidant capacity of PYC by the trolox equivalent antioxidant capacity (TEAC) assay and the cytotoxicity by neutral red uptake (NRU) test in Chinese Hamster Ovary (CHO) cells. The genotoxic and antigenotoxic effects of PYC were evaluated by the cytokinesis-blocked micronucleus (CBMN) and alkaline comet assays in human peripheral blood lymphocytes. At the concentrations of 2-200 µg/ml, PYC was found to have antioxidant activity. The viability of CHO cells during 24h exposure were not affected at the concentrations of 5-150 µg/ml of PYC. IC50 value of PYC was found to be 285 µg/ml. At the concentrations above 100 µg/ml, PYC alone induced DNA damage and increased MN frequency, although PYC at all concentrations in a dose dependent manner revealed a reduction in the frequency of MN and the extent of DNA damage induced by H2O2. These results suggest PYC might reduce H2O2 induced chromosome breakage and loss and DNA damage in cultured human lymphocytes.

Protective effects of curcumin against oxidative stress parameters and DNA damage in the livers and kidneys of rats with biliary obstruction.

Curcumin, a most active antioxidant compound, has been suggested to have potential beneficial effects against most metabolic and psychological disorders, including cholestasis. In the present study, the effects of curcumin against oxidative stress and DNA damage induced by bile duct ligation (BDL) in Wistar albino rats for 14 days were investigated. The rats were divided into three following groups: Sham group, the BDL group and the BDL+curcumin group. A daily dose of 50mg/kg curcumin was given to the BDL+curcumin group intragastrically for 14 days. The biomarkers of hepatocellular damage were decreased in the BDL+curcumin group compared to the BDL group, indicating that curcumin recovered the liver functions. DNA damage as assessed by the alkaline comet assay was also found to be low in the BDL+curcumin group. Curcumin significantly reduced malondialdehyde and nitric oxide levels, and enchanced reduced glutathione levels and catalase, superoxide dismutase, and glutathione S-transferase enzymes activities in the livers and kidneys of BDL group. Curcumin treatment in BDL group was found to decrease tumor necrosis factor-alpha levels in the livers of rats. These results suggest that curcumin might have protective effects on the cholestasis-induced injuries in the liver and kidney tissues of rats.