Vanillin attenuates CdCl2-induced cytotoxicity in isolated human erythrocytes.

Cadmium is a toxic heavy metal with no physiological role in the human body. Cadmium has high mobility due to its widespread industrial use, with no safe and effective therapeutic management. Cadmium toxicity manifests by increasing oxidative stress in target cells. We have explored the potential role of vanillin, a plant phenolic aldehyde and antioxidant, in mitigating cadmium chloride (CdCl2) induced hemotoxicity using isolated human erythrocytes. CdCl2 was added to erythrocytes, in the absence and presence of vanillin. Incubation of erythrocytes with CdCl2 alone inhibited methemoglobin reductase and enhanced methemoglobin level. Heme degradation and release of free iron (Fe2+), along with protein and membrane lipid oxidation, were also increased. A CdCl2-induced enhancement in reactive oxygen and nitrogen species was also seen, lowering the overall antioxidant power of cells. However, pre-incubation of erythrocytes with vanillin resulted in significant decreased generation of reactive species and prevented heme degradation and heme oxidation. Vanillin augmented the erythrocyte antioxidant capacity and reinstated the activities of major antioxidant, plasma membrane-bound and glucose metabolism enzymes. Scanning electron microscopy showed that CdCl2 treatment led to the formation of echinocytes which was prevented by vanillin. In all cases, no harmful effects of vanillin alone were seen. Thus, vanillin alleviates the toxicity of cadmium and can be potentially employed as a chemoprotectant against the damaging effects of this heavy metal.

Mancozeb-induced cytotoxicity in human erythrocytes: enhanced generation of reactive species, hemoglobin oxidation, diminished antioxidant power, membrane damage and morphological changes.

Mancozeb is an ethylene bis-dithiocarbamate fungicide extensively used in agriculture to safeguard crops from various fungal diseases. The general population is exposed to mancozeb through consumption of contaminated food or water. Here, we have investigated the effect of mancozeb on isolated human erythrocytes under in vitro conditions. Erythrocytes were treated with different concentrations of mancozeb (0, 5, 10, 25, 50, 100 µM) and incubated for 24 h at 37 °C. Analysis of biochemical parameters and cell morphology showed dose-dependent toxicity of mancozeb in human erythrocytes. Mancozeb treatment caused hemoglobin oxidation and heme degradation. Protein and lipid oxidation were enhanced, while a significant decrease was seen in reduced glutathione and total sulfhydryl content. A significant increase in the generation of reactive oxygen and nitrogen species was detected in mancozeb-treated erythrocytes. The antioxidant capacity and the activity of key antioxidant enzymes were greatly diminished, while crucial metabolic pathways were inhibited in erythrocytes. Damage to the erythrocyte membrane on mancozeb treatment was apparent from increased cell lysis and osmotic fragility, along with the impairment of the plasma membrane redox system. Mancozeb also caused morphological alterations and transformed the normal discoid-shaped erythrocytes into echinocytes and stomatocytes. Thus, mancozeb induces oxidative stress in human erythrocytes, impairs the antioxidant defense system, oxidizes cellular components, that will adversely affect erythrocyte structure and function.

Cadmium chloride generates cytotoxic reactive species that cause oxidative damage and morphological changes in human erythrocytes.

Cadmium chloride (CdCl2) is a widely used industrial compound that exhibits multiple organ toxicity. Cadmium is transported through blood where erythrocytes are exposed to its action. Here the effect of CdCl2 on human erythrocytes was examined under in vitro conditions. Human erythrocytes were treated with 0.01-0.5 mM CdCl2 for 24 h at 37 °C. Lysates were made from CdCl2 treated and untreated (control) cells and used for further analysis. CdCl2 treatment resulted in marked hemolysis of erythrocytes and oxidation of hemoglobin to methemoglobin. This will result in anemia and also reduce the oxygen carrying ability of erythrocytes. Hemoglobin oxidation was accompanied by degradation of heme and release of free ferrous iron moiety. Further analysis showed elevated lipid hydroperoxides and formation of advanced oxidation protein products along with reduction in total sulfhydryl content, indicating the generation of oxidative stress condition in the cell. Incubation of erythrocytes with CdCl2 enhanced generation of reactive oxygen and nitrogen species, decreased the antioxidant power and inhibited pathways of glucose metabolism. Plasma membrane was damaged as indicated by enhanced osmotic fragility and inhibition of membrane bound enzymes. This was confirmed by electron microscopy which showed formation of echinocytes. These results show that CdCl2 generates reactive species which impair the antioxidant system resulting in oxidative damage to erythrocytes.

Interaction of mancozeb with human hemoglobin: Spectroscopic, molecular docking and molecular dynamic simulation studies.

Mancozeb is a broad-spectrum fungicide used extensively in agriculture to protect plants from numerous diseases. Hemolysis of human erythrocytes on exposure to mancozeb has been reported. In the present study, we investigated the interaction of mancozeb with human hemoglobin (Hb) using multi-spectroscopic techniques, molecular docking and molecular dynamic simulation. UV-visible spectroscopy studies suggested intimate binding of mancozeb to Hb. Mancozeb quenched the intrinsic fluorescence of Hb and Stern-Volmer plots revealed that the quenching mechanism was of static type. Evaluation of thermodynamic parameters indicated that the binding of Hb to mancozeb was spontaneous, with van der Waals forces and hydrogen bonding being the key contributors in the binding reaction. Synchronous fluorescence experiments demonstrated that mancozeb altered the microenvironment around tryptophan residues, whereas polarity around tyrosine residues was not changed. Circular dichroism studies showed a decrease in the α helical content of Hb upon interaction with mancozeb. The inhibition of esterase activity showed that mancozeb can impair the enzymatic functions of Hb. Molecular docking study revealed that strong binding affinity existed between mancozeb and Hb, with hydrophobic forces playing a crucial role in the interaction. Molecular dynamic simulation showed that mancozeb formed a stable complex with Hb resulting in slight unfolding of the protein. To sum up, the results of this study show that mancozeb binds strongly to Hb, induces conformational changes in Hb and adversely affects its function.

Oral administration of thiram inhibits brush border membrane enzymes, oxidizes proteins and thiols, impairs redox system and causes histological changes in rat intestine: A dose dependent study.

Pesticides are extensively employed worldwide, especially in agriculture to control weeds, insect infestation and diseases. Besides their targets, pesticides can also affect the health of non-target organisms, including humans The present study was conducted to study the effect of oral exposure of thiram, a dithiocarbamate fungicide, on the intestine of rats. Male rats were administered thiram at doses of 100, 250, 500 and 750 mg/kg body weight for 4 days. This treatment reduced cellular glutathione, total sulfhydryl groups but enhanced protein carbonyl content and hydrogen peroxide levels. In addition, the activities of all major antioxidant enzymes (catalase, thioredoxin reductase, glutathione peroxidase and glutathione-S-transferase) except superoxide dismutase were decreased. The antioxidant power of the intestine was impaired lowering the metal-reducing and free radical quenching ability. Administration of thiram also led to inhibition of intestinal brush border membrane enzymes, alkaline phosphatase, γ-glutamyl transferase, leucine aminopeptidase and sucrase. Activities of enzymes of pentose phosphate pathway, citric acid cycle, glycolysis and gluconeogenesis were also inhibited. Histopathology showed extensive damage in the intestine of thiram-treated rats at higher doses. All the observed effects were in a thiram dose-dependent manner. The results of this study show that thiram causes significant oxidative damage in the rat intestine which is associated with the marked impairment in the antioxidant defense system.

Therapeutic effect of vitamin B3 on hyperglycemia, oxidative stress and DNA damage in alloxan induced diabetic rat model.

Evidences in the form of experimental analysis and scientific investigations suggest that oxidative stress embody an imperative role in the onset and progression of type-2 diabetes mellitus (T2DM). Aberrant elevation in levels of free radicals, as observed upon disease onset, and the subsequent reduction in anti-oxidant defenses is pernicious to metabolic enzymes and cellular organelles. Niacin (Vitamin B3) is an essential nutrient for humans and is considered to be an important food additive for animals too. This research was conducted to examine the effect of nutraceutical antioxidant on diabetic environment. This important member of Vitamin B complex is a forerunner of nicotinamide adenine dinucleotide (NAD) and also nicotinamide adenine dinucleotide phosphate (NADP), both of them serving as coenzymes for several metabolic enzymes. This study reports the effects of niacin supplementation in alloxan induced diabetic rats divided into five groups. Diabetes induced rats were further treated with niacin at two doses (10 and 15 mg /kg body weight) and compared with a control set of diabetes without treatment. Niacin treatment showed recovery in almost all parameters in a dose reliant pattern. A notable decline in oxidative stress parameters with reductions in fasting blood glucose levels was observed. Histological studies reveal damage recovery in the liver as well as kidney tissues. A notable amount of recovery was observed in cellular DNA damage. As a deduction, it is advocated that dietary niacin supplementation might help in reducing problems associated with diabetes. A probable mechanism pertaining to the action of niacin is proposed as well.