Reduction In Adenosine Triphosphate Content Research Articles

Honokiol attenuates oxidative stress-induced cytotoxicity in human keratinocytes via activating AMPK signaling

Objective: To investigate the effect of honokiol on oxidative damage in HaCaT human keratinocytes. Methods: HaCaT cells were exposed to hydrogen peroxide (H2O2), following pretreatment with various concentrations of honokiol. The alleviating effects of honokiol on HaCaT cell viability and cell death, reactive oxygen species (ROS) production, DNA damage, mitochondrial dynamics, and inhibition of adenosine triphoaphate production against H2O2 were investigated. Western blotting analysis was used to analyze the expression levels of specific proteins. Results: Honokiol suppressed H2O2-induced cytotoxicity and DNA damage by blocking abnormal ROS accumulation. Honokiol also prevented apoptosis by inhibiting loss of mitochondrial membrane potential and release of cytochrome c from the mitochondria into the cytosol, decreasing the Bax/Bcl-2 ratio, and reducing the activity of caspase-3 in H2O2-stimulated HaCaT cells. In addition, honokiol attenuated H2O2-induced reduction of adenosine triphosphate content, and activation of AMP-activated protein kinase (AMPK) was markedly promoted by honokiol in H2O2-stimulated cells. Importantly, the anti-apoptosis and anti-proliferative activity of honokiol against H2O2 was further enhanced by adding an activator of AMPK, indicating that honokiol activated AMPK in HaCaT keratinocytes to protect against oxidative damage. Conclusions: The present results indicate that honokiol may be useful as a potential therapeutic agent against various oxidative stress-related skin diseases.

MiR-423-5p inhibition alleviates cardiomyocyte apoptosis and mitochondrial dysfunction caused by hypoxia/reoxygenation through activation of the wnt/β-catenin signaling pathway via targeting MYBL2.

MicroRNA (miR) plays an integral role in cardiovascular diseases. M-iR-423-5p is aberrantly expressed in patients with myocardial infarction and heart failure. The aim of the present study was to study the roles and mechanisms of miR-423-5p in hypoxia/reoxygenation (H/R) mediated cardiomyocytes injury. H9C2 cells were transfected with negative control, miR-423-5p mimic, and inhibitor for 48 hr, followed by exposed to H/R condition. Cell apoptosis rate, caspase 3/7 activities, Bax and cleaved-caspase 3 (c-caspase 3) protein levels were assayed by flow cytometry, Caspase-Glo 3/7 Assay kit, western blot analysis, respectively. Furthermore, the mitochondrial membrane potential, adenosine triphosphate (ATP)content, reactive oxygen species (ROS) production, and Drp1 expression were also investigated. Furthermore, the dual-luciferase reporter assay was used to evaluate the relationship between miR-423-5p and Myb-related protein B (MYBL2). The roles of miR-423-5p in wnt/β-catenin were assessed by western blot analysis. The results revealed that H/R triggered miR-423-5p expression. Overexpression of miR-423-5p promoted cardiomyocyte apoptosis, enhanced the activities of caspase 3/7, upregulated the expression of Bax and c-caspase 3. miR-423-5p upregulation caused the loss of mitochondrial membrane potential and the reduction of ATP content, the augment of ROS production and Drp1 expression. However, the opposite trends were observed upon suppression of miR-423-5p. In addition, miR-423-5p could target the 3' untranslated region of MYBL2. miR-423-5p depletion led to the activation of the wnt/β-catenin signaling pathway via targeting MYBL2. Knockdown of MYBL2 was obviously reversed the roles of miR-423-5p in apoptosis and mitochondrial dysfunction. Taken together, miR-423-5p suppression reduced H/R-induced cardiomyocytes injury through activation of the wnt/β-catenin signaling pathway via targeting MYBL2 in cardiomyocytes.

Efficacy of β-phenylethylamine as a novel anti-microbial and application as a liquid catheter flush.

With this study, we introduce a liquid flush for catheters and other tubing-based applications that consists of a solution of β-phenylethylamine (PEA) in tryptic soy broth. The initial experiments in multiwell polystyrene plates were conducted with Escherichia coli K-12 to assess the effectiveness of PEA at reducing planktonic growth, as well as the biomass and adenosine triphosphate (ATP) content of biofilm; PEA reduced these growth parameters as a function of increasing concentration. This effect was also seen in mutants of PEA catabolism, which leads us to believe that the PEA effect is due to PEA itself and not one of its degradation products. Since PEA reduced planktonic growth and biofilm when added at the time of inoculation, as well as at later time points, we propose PEA as a novel compound for the prevention and treatment of biofilm. PEA reduced planktonic growth and the ATP content of the biofilm for five bacterial pathogens, including an enterohemorrhagic E. coli, two uropathogenic E. coli, Pseudomonas aeruginosa and Staphylococcus aureus. A major finding of this study is the reduction of the ATP content of biofilm that formed in silicone tubing by periodic flushes of PEA. This experiment was performed to model antibiotic-lock treatment of an intravenous catheter. It was found that 10 mg ml-1 of PEA reduced the ATP content of biofilm of five bacterial strains by 96.3 % or more after 2 weeks of incubation and three treatments with PEA. For P. aeruginosa, the reduction in ATP content was paralleled by an identical percentage reduction in viable cells in the biofilm.

Atrazine exposure causes mitochondrial toxicity in liver and muscle cell lines.

Objective:Chronic exposure to atrazine and other pesticides is reported to cause metabolic disorders, yet information on effects of atrazine on expression of genes relevant to mitochondrial function is largely missing. In the present study, therefore, we investigated the expression of a battery of nuclear- and mitochondrial-encoded genes involved in oxidative phosphorylation (OXPHOS) in human liver (HepG2) and rat muscle (L6) cell lines due to short-term atrazine exposure.Materials and Methods:We have determined the EC50 values of atrazine for cytotoxicity and mitochondrial toxicity (mitotoxicity) in terms of adenosine triphosphate (ATP) content in HepG2 and L6 cells. Further, the mRNA expression of nuclear- and mitochondrial-encoded genes was analyzed using quantitative real-time polymerase chain reaction.Results:The EC50 value of atrazine for mitotoxicity in HepG2 and L6 cells was found to be about 0.162 and 0.089 mM, respectively. Mitochondrial toxicity was indicated by reduction in ATP content following atrazine exposure. Atrazine exposure resulted in down-regulation of many OXPHOS subunits expression and affected biogenesis factors’ expression. Most prominently, superoxide dismutase (SOD) and sirtuin 3 (SIRT3) expressions were up-regulated in HepG2 cells, whereas SIRT3 expression was alleviated in L6 cells, without significant changes in SOD levels. Mitochondrial transcription factor A (TFAM) and SIRT1 expression were significantly down-regulated in both cell lines.Conclusion:Results suggest that TFAM and SIRT1 could be involved in atrazine-induced mitochondrial dysfunction, and further studies can be taken up to understand the mechanism of mitochondrial toxicity. Further study can also be taken up to explore the possibility of target genes as biomarkers of pesticide toxicity.

The Protection of Selenium on Adriamycin-Induced Mitochondrial Damage in Rat

Although adriamycin (ADR) exhibits high anti-tumor efficacy in vitro, its clinical use in cancer chemotherapy is limited due to its high renal toxicity. This study investigated the mechanism of ADR nephropathy and the protective effect of selenium on ADR-induced kidney damage by analyzing of the relationship between selenium and mitochondria. Rats were divided into four groups. The first group was injected with saline i.p. for 21 days, the second group received the 4 mg/kg i.p. ADR every alternate day for 8 days, the third group received the 50 μg/kg i.p. Se for 21 days, and the fourth group received the Se. ADR co-administration i.p. blood pressures were assessed, the mitochondrial membrane potential (MMP) was assessed, and the adenosine triphosphate (ATP) levels were determined. The total antioxidant (TAS) and oxidant status (TOS) in cytosol, the mitochondria of kidney cells, and plasma were measured. Mitochondrial TAS decreased and TOS increased in the ADR group compared to the Se group. ADR-treated rats showed significantly lower MMP than did the control and Se groups. MMP was significantly restored in the Se + ADR group through selenium treatment compared to the ADR group (p < 0.01). In the ADR group, a reduction in ATP content was seen compared to the control and Se groups (p < 0.01). ATP level was significantly restored through treatment with selenium in the Se + ADR group compared to the ADR group (p < 0.01). We concluded that selenium is effective in vivo against ADR-induced kidney damage via the restoration of TAS and TOS, which prevented mitochondrial damage.

Leucocyte filtration of salvaged blood during cardiac surgery: effect on red blood cell function in concentrated blood compared with diluted blood

Leucocyte filtration of salvaged blood has been suggested to prevent patients from receiving activated leucocytes during auto-transfusion in cardiac surgery. This study examines whether leucocyte filtration of salvaged blood affects the red blood cell (RBC) function and whether there is a difference between filtration of the concentrated and diluted blood on RBC function. Forty patients undergoing cardiac surgery with cardiopulmonary bypass were randomly divided into a group receiving leucocyte filtration of concentrated blood (High-Hct, n=20) and another group receiving leucocyte filtration of the diluted blood (Low-Hct, n=20). During operation, all the salvaged blood, as well as the residual blood, from the heart-lung machine was filtered. In the High-Hct group, blood was concentrated with a cell saver prior to filtration, whereas in the Low-Hct group, blood was filtered without concentration. RBC function was represented by RBC aggregation and deformability measured by a laser-assisted optical rotational cell analyser and by the RBC 2,3-diphosphoglycerate (2,3-DPG) and adenosine triphosphate (ATP) contents with conventional biochemical tests. Leucocyte filtration of diluted blood with a low haematocrit (14+/-4%) did not affect RBC function. However, when the concentrated blood with a high haematocrit (69+/-12%) was filtered, there was a reduction of ATP content in RBCs after passing through the filter (from 1.45+/-0.57 micromol g(-1) Hb to 0.92+/-0.75 micromol g(-1) Hb, p<0.05). For patients who received the concentrated blood, their in vivo RBC function did not differ from those who received diluted blood. Leucocyte filtration of the diluted salvaged blood during cardiac surgery does not affect RBC function, but it tends to deplete the ATP content of RBCs as the salvaged blood has been concentrated prior to filtration.

Oral Administration of Amino Acidic Supplements Improves Protein and Energy Profiles in Skeletal Muscle of Aged Rats: Elongation of Functional Performance and Acceleration of Mitochondrial Recovery in Adenosine Triphosphate After Exhaustive Exertion

Sarcopenia is an inevitable age-related degenerative process chiefly characterized by decreased synthesis of muscle proteins and impaired mitochondrial function, leading to progressive loss of muscle mass. Here, we sought to probe whether long-term administration of oral amino acids (AAs) can increase protein and adenosine triphosphate (ATP) content in the gastrocnemius muscle of aged rats, enhancing functional performance. To this end, 6- and 24-month-old male Fisher 344 rats were divided into 3 groups: group A (6-month-old rats) and group B (24-month-old rats) were used as adult and senescent control group, respectively, while group C (24-month-old rats) was used as senescent treated group and underwent 1-month oral treatment with a mixture of mainly essential AAs. Untreated senescent animals exhibited a 30% reduction in total and fractional protein content, as well as a 50% reduction in ATP content and production, compared with adult control rats (p <0.001). Long-term supplementation with mixed AAs significantly improved protein and high-energy phosphate content, as well as the rate of mitochondrial ATP production, conforming their values to those of adult control animals (p <0.001). The improved availability of protein and high-energy substrates in the gastrocnemius muscle of treated aged rats paralleled a significant enhancement in functional performance assessed by swim test, with dramatic elongation of maximal exertion times compared with untreated senescent rats (p <0.001). In line with these findings, we observed that, after 6 hours of rest following exhaustive swimming, the recovery in mitochondrial ATP content was approximately 70% in adult control rats, approximately 60% in senescent control rats, and normalized in treated rats as compared with animals of the same age unexposed to maximal exertion (p <0.001). In conclusion, nutritional supplementation with oral AAs improved protein and energy profiles in the gastrocnemius of treated rats, enhancing functional performance and accelerating high-energy phosphate recovery after exhaustive exertion.

Effect of melatonin administration on parameters related to oxidative damage in hepatocytes isolated from old Wistar rats

Aging induces changes in several organs and tissues, such as the liver, and this process might be due to oxidative damage caused by free radicals and inflammatory mediators. Melatonin is a secretory product with well-known antioxidant properties. The aim of this study was to investigate the effect of melatonin administration on age-induced alterations in hepatocytes. Twenty-two-month old male Wistar rats were treated with oral melatonin for 10 wk. At the end of the treatment, hepatocytes were isolated and cultured, and different parameters were measured in both cells and medium. Aging induced a significant increase in lipid peroxidation, nitric oxide, carbon monoxide and cyclic guanosyl-monophosphate, as well as a reduction in adenosine triphosphate content and phosphatidylcholine synthesis when compared to young animals. Melatonin administration significantly ameliorated all these age-related changes in males. Melatonin administration seems to exert beneficial effects against age-induced changes in hepatocytes.

Alcohol-induced liver injury in mice lacking Cu, Zn-superoxide dismutase

Because alcoholic liver disease has been linked to oxidative stress, we investigated the effect of a compromised antioxidant defense system, Cu, Zn-superoxide dismutase (Sod1) deficiency, on alcohol-induced liver injury. C57BL/129SV wild-type (Sod1(+/+)) and Sod1 knockout (Sod1(-/-)) mice were fed dextrose or ethanol (10% of total calories) liquid diets for 3 weeks. Histologic evaluation of liver specimens of Sod1(-/-) mice fed ethanol showed the development of liver injury ranging from mild to extensive centrilobular necrosis and inflammation. Sod1(+/+) mice fed ethanol showed mild steatosis; both Sod1(+/+) and Sod1(-/-) mice fed the dextrose diet had normal histology. Alanine transaminase levels were significantly elevated only in Sod1(-/-) mice fed ethanol. Cytochrome P450 2E1 (CYP2e1) activity was elevated about 2-fold by ethanol in Sod1(+/+) and Sod1(-/-) mice. Ethanol consumption increased levels of protein carbonyls and lipid peroxidation aldehydic products in the liver of Sod1(-/-) mice. Hepatic adenosine triphosphate (ATP) content was reduced dramatically in Sod1(-/-) mice fed ethanol in association with a decrease in the mitochondrial reduced glutathione (GSH) level and activity of MnSOD. Immunohistochemical determination of 3-nitrotyrosine (3NT) residues in liver sections of the Sod1 knockout mice treated with ethanol showed a significant increase of 3NT staining in the centrilobular areas. In conclusion, a rather moderate ethanol consumption promoted oxidative stress in Sod1(-/-) mice, with increased formation of peroxynitrite, protein carbonyls, and lipid peroxidation and decreased mitochondrial GSH and MnSOD. We speculate that the increased oxidative stress causes mitochondrial damage and reduction of ATP content, leading to alcoholic liver injury. This model may be useful in further mechanistic studies on alcohol-induced liver injury.

Neutrophil function in hyperosmotic NaCl is preserved by phosphoenol pyruvate.

Hyperosmolarity in the urinary tract inhibits the host defense against bacterial infection. NaCl contributes most to osmolarity in the renal medulla and urine. Therefore, we studied the effect of hyperosmolar NaCl on neutrophil function. When osmolarity was increased by NaCl, polymorphonuclear leukocytes (PMNL) became defective in phagocytosis, intracellular killing of bacteria, chemotactic activity, and superoxide production. This coincided with a decrease in the intracellular adenosine triphosphate (ATP) content of PMNL. Both the inhibition of superoxide production and the reduction in ATP content did not occur at 4 degrees C. By increasing ATP content, phosphoenol pyruvic acid (PEP) protected against the decrease in superoxide production. These results suggest that leukocyte function is inhibited by high concentrations of NaCl due to the activation of the Na(+)-K+ pump. PEP, an ATP precursor, can protect PMNL against osmotic stress by raising the intracellular concentration of ATP.

Contractile failure and high-energy phosphate turnover during hypoxia: 31P-NMR surface coil studies in living rat.

Cardiac failure appears rapidly during severe hypoxia and precedes a substantial reduction in adenosine triphosphate content. Reduced adenosine triphosphate turnover, in the presence of nearly normal content, may be the metabolic basis for contractile failure during hypoxia. To measure both the myocardial content and the turnover rates of high-energy phosphate compounds during hypoxia, we performed 31P-nuclear magnetic resonance studies by placing a surface coil directly over the left ventricle in intubated rats that were instrumented for hemodynamic measurements and ventilated with either 21, 10, or 8% O2. Normoxia produced a hemodynamic and metabolic steady state for 4 hours and hypoxia for at least 60 minutes. Under normoxic ventilation (n = 10, mean +/- SD), the arterial PO2 was 96 +/- 14, pH 7.38 +/- 0.11, and systolic blood pressure 96 +/- 8 mm Hg; under hypoxic ventilation with 10% O2 (n = 5), the arterial PO2 was 57 +/- 10, pH 7.39 +/- 0.09, and systolic pressure 68 +/- 10; and under hypoxic ventilation with 8% O2 (n = 5), the PO2 was 52 +/- 7, pH 7.37 +/- 0.04, and systolic pressure 51 +/- 4. Hypoxic ventilation with 10 or 8% O2 decreased the creatine phosphate content from 51.4 +/- 5.4 mumol/g dry wt to 39.3 +/- 5.4 and 45.6 +/- 4.1 and depressed adenosine triphosphate slightly from 25.0 mumol/g dry wt to 21.8 +/- 2.1 and 21.9 +/- 1.0, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

High-energy phosphate metabolism of isolated coronary arteries in the dog.

The correlation between phosphate metabolism and isometric contraction was investigated in helical strips from dog coronary artery. Adenosine triphosphate (ATP) and creatine phosphate (CP) contents of resting strips did not significantly change during anoxia. ATP content remained unchanged during potassium-contracture under aerobic conditions but was significantly reduced by anoxia. Pretreatment with iodoacetate resulted in a striking reduction in ATP content with a less marked decrease in CP content. ATP-depleting effect of iodoacetate was reduced slightly by the addition of pyruvate, but contractile response was not restored. It is considered that high-energy phosphates provided by anaerobic glycolysis can meet the energy requirements of resting coronary vessels but are insufficient for maintaining active tension. In resting strips equilibrated for 2 h in oxygenated bath solution, the reactivity to KCl and anoxia appears to be dependent on the availability of ATP.