Liver Mitochondria Research Articles

Hepatotoxic doses of copper sulfate induce metabolic memory in the redox system, which has an age-dependent nature

Abstract. We tested the hypothesis that ageing is a consequence of the formation of metabolic memory and the possible role of the redox system as a basic, evolutionarily ancient system of metabolism regulation in stable metabolic patterns formation or metabolic states chronisation changed during adaptation. Three sequential intraperitoneal administration of copper sulfate to young (3-month-old Wistar rats) and old animals (20 months) at a dose of 1 mg/100 g of body weight (33% of the lethal dose) were used as adaptive inducers of the redox system. The amount of lipid hydroperoxides in mitochondria, cytosol in liver cells and blood serum, the activity of mitochondrial aconitate hydratase as an indicator of oxidative stress and the activity of a number of antioxidant enzymes were determined to assess the initial metabolic states, i. e. before exposure and 1, 30 and 60 days after exposure to copper sulfate on the body. It was shown that the amount of lipid hydroperoxides (LOOHs) in the liver mitochondria and blood serum of old rats before exposure to copper sulfate was more than 30% lower than in young animals, while the aconitase activity (an indicator of oxidative stress) was the same in animals of these ages. A lower amount of LOOHs coincided with an increased glutathione peroxidase activity in old animals. In old rats, the increased amount of LOOHs induced by copper ions was preserved even after eliminating copper from the body 30 days after administration. At the same time, it was restored to the original level in the young animals. The glutathione peroxidase and aconitase activity in mitochondria remained below the control values even after the inducer elimination, and this was age-depending. The obtained results do not contradict the hypothesis of metabolic memory's role in ageing mechanisms. We postulate a relationship between the duration of maintenance of altered metabolic patterns and the polyfunctionality of enzymes and other metabolites. _________________________________________________________________________________________ Keywords: ageing; redox system; metabolic memory; lipid hydroperoxides; antioxidant enzymes; chronic states.

Comparison of Mitochondrial and Antineoplastic Effects of Amiodarone and Desethylamiodarone in MDA-MB-231 Cancer Line.

Previously, we have demonstrated that amiodarone (AM), a widely used antiarrhythmic drug, and its major metabolite desethylamiodarone (DEA) both affect several mitochondrial processes in isolated heart and liver mitochondria. Also, we have established DEA's antitumor properties in various cancer cell lines and in a rodent metastasis model. In the present study, we compared AM's and DEA's mitochondrial and antineoplastic effects in a human triple-negative breast cancer (TNBC) cell line. Both compounds reduced viability in monolayer and sphere cultures and the invasive growth of the MDA-MB-231 TNBC line by inducing apoptosis. They lowered mitochondrial trans-membrane potential, increased Ca2+ influx, induced mitochondrial permeability transition, and promoted mitochondrial fragmentation. In accordance with their mitochondrial effects, both substances massively decreased overall, and even to a greater extent, mitochondrial ATP production decreased, as determined using a Seahorse live cell respirometer. In all these effects, DEA was more effective than AM, indicating that DEA may have higher potential in the therapy of TNBC than its parent compound.

CYB5R3 overexpression exhibits sexual dimorphism: Mitochondrial and metabolic adaptations in transgenic female mice during calorie restriction

There is a pressing need to develop new strategies for enhancing health in the elderly and preventing the rise in age-related diseases. Calorie restriction without malnutrition (CR) stands among the different antiaging interventions. Lifelong CR leads to increased expression and activity of plasma membrane CYB5R3, and male mice overexpressing CYB5R3 exhibit some beneficial adaptations that are also seen with CR. However, the mechanisms involved in both interventions could be independent since key aspects of energy metabolism and tissue lipid profile do not coincide, and many of the changes induced by CR in mitochondrial abundance and dynamics in the liver and skeletal muscle could be counteracted by CYB5R3 overexpression. In this study, we sought to elucidate the impact of CR on key markers of metabolic status, mitochondrial function, and pro-oxidant/antioxidant balance in transgenic (TG) female mice overexpressing CYB5R3 compared to their WT littermates. In females fed ad libitum, CYB5R3 overexpression decreased fat mass, led to a preferred utilization of fatty acids as an energy source, upregulated key antioxidant enzymes, and boosted respiration both in skeletal muscle and liver mitochondria, supporting that CYB5R3 overexpression is phenotypic closer to CR in females than in males. Whereas some markers of mitochondrial biogenesis and dynamics were found decreased in TG females on CR, as also found for the levels of Estrogen Receptor α, mitochondrial abundance and activity were maintained both in skeletal muscle and in liver. Our results reveal overlapping metabolic adaptations resulting from the overexpression of CYB5R3 and CR in females, but a specific crosstalk occurs when both interventions are combined, differing from the adaptations observed in TG males.

Adaptive Effects of Intermittent Hypoxia Training on Oxygen-Dependent Processes as a Potential Therapeutic Strategy Tool

Background/Aims: Important benefits of intermittent hypoxic training (IHT) have emerged as an effective tool for enhancing adaptive potential in different pathological states, among which acute hypoxia dominates. Therefore, the aim of our study was to evaluate the mechanisms related to the effects of the nitric oxide system (nitrites, nitrates, carbamide, and total polyamine content) on ADP-stimulated oxygen consumption and oxidative phosphorylation in heart and liver mitochondria and biomarkers of oxidative stress in the blood, heart, and liver of rats exposed to the IHT method and acute hypoxia and treated with the amino acid L-arginine (600 mg/kg, 30 min) or the NO synthase inhibitor L-NNA (35 mg/kg, 30 min) prior to each IHT session. Methods: We analysed the modulation of the system of oxygen-dependent processes (mitochondrial respiration with the oxygraphic method, microsomal oxidation, and lipoperoxidation processes using biochemical methods) in tissues during IHT in the formation of short-term and long-term effects (30, 60, and 180 days after the last IHT session) with simultaneous administration of L-arginine. In particular, we investigated how mitochondrial functions are modulated during intermittent hypoxia with the use of oxidation substrates (succinate or α-ketoglutarate) in bioenergetic mechanisms of cellular stability and adaptation. Results: The IHT method is associated with a significant increase in the production of endogenous nitric oxide measured by the levels of its stable metabolite, nitrite anion, in both plasma (almost 7-fold) and erythrocytes (more than 7-fold) of rats. The intensification of nitric oxide-dependent pathways of metabolic transformations in the energy supply processes in the heart and liver, accompanied by oscillatory mechanisms of adaptation in the interval mode, causes a probable decrease in the production of urea and polyamines in plasma and liver, but not in erythrocytes. The administration of L-arginine prior to the IHT sessions increased the level of the nitrite-reducing component of the nitric oxide cycle, which persisted for up to 180 days of the experiment. Conclusion: Thus, the efficacy of IHT and its nitrite-dependent component shown in this study is associated with the formation of long-term adaptive responses by preventing the intensification of lipoperoxidation processes in tissues due to pronounced changes in the main enzymes of antioxidant defence and stabilisation of erythrocyte membranes, which has a pronounced protective effect on the system of regulation of oxygen-dependent processes as a whole.

Вплив етанолу та циклоспорину А на дихання ізольованих гепатоцитів щурів

It is known that ethanol affects the oxidative processes in liver mitochondria, which can be one of the mechanisms of the development of the alcoholic liver disease. A well-known immunosuppressant cyclosporine A is also an inhibitor of mitochondrial permeabi­lity transition pore. However, the pharmacological interaction of ethanol and cyclosporin A is not sufficiently studied. The aim of the study was to investigate the effect of ethanol and cyclosporine A on the respiration rate of isolated rat hepatocytes. Five male Wistar rats were used in the study. Hepatocytes were isolated by perfusion with collagenase. Cell plasma membrane integrity was assessed with trypan blue staining (0.1 %). Intact cell number was 85.7±0.92 %. The rate of oxygen consumption was measured using a polarographic device based on a Clark electrode. Protonophore FCCP was used to study the uncoupled respiration. Cell respiration was studied in the presence of glucose, pyruvate or monomethylsuccinate in solution. Statistical analysis was performed with Origin Pro 2018 software. Significance of difference between groups was evaluated with analysis of variance test. After one hour incubation with ethanol (50 mM), the rate of basal respiration of hepatocytes oxidizing glucose increased by a meager 8 %. Similar result was obtained upon the presence of pyruvate in solution. Monomethylsuccinate, however, abolished the effect of ethanol on basal respiration. he but not uncoupled respiration of hepatocytes upon glucose or pyruvate oxidation. Ethanol did not affect the uncoupled respiration of hepatocytes in presence of glucose, pyruvate of monomethylsuccinate. Incubation with cyclocporin A did not cause any changes in cell oxygen consumption in all experiments. Cyclosporin A also did not modify the effects of ethanol on basal respiration of hepatocytes. Therefore, no pharmacological no interaction between ethanol and cyclosporin A was detected, which could be evidenced by the change of mitochondrial respiration of isolated hepatocytes.

PGC-1α loss promotes mitochondrial protein lactylation in acetaminophen-induced liver injury via the LDHB-lactate axis

Coronavirus disease 2019 (COVID-19) affected people worldwide, and fever is one of the major symptoms of this disease. Although Acetaminophen (APAP) is a common fever-reducing medication, it can also mediate liver injury. However, the role of PGC-1α in regulating mitochondrial quality control by lactate dehydrogenase B (LDHB), a vital enzyme catalyzing the conversion of lactate to pyruvate, in APAP-induced hepatotoxicity, is unclear. Here, gene expression omnibus data of patients with APAP-induced liver injury were used to explore gene expression profiles. AML12 cells and C57/BL6 mice were used to establish models of APAP-induced acute liver injury. SIRT1 and PGC-1α were overexpressed in vitro via lentiviral transfection to establish stable cell lines. The results showed that APAP treatment decreased SIRT1/PGC-1α/LDHB expression and increased protein lactylation, mitochondrial lactate levels, and pathological damage in liver mitochondria. PGC-1α upregulation or activation ameliorated APAP-induced damage in the cells and liver. Furthermore, PGC-1α overexpression increased LDHB synthesis, reduced lactylation, and induced a switch from lactate to pyruvate production. These results suggest that PGC-1α and LDHB play a role in APAP-induced liver injury by regulating mitochondrial quality control and lactate metabolic reprogramming. Therefore, the PGC-1α/LDHB axis is a potential therapeutic target for APAP-induced liver injury.

Low Hepatic Cardiolipin Levels Promote Increased Mitochondrial Electron Leak and Metabolic Dysfunction-associated Steatohepatitis

Mitochondria adapt to changing energetic demands of the cell. We recently reported that mitochondrial membrane lipid composition represents a regulatory mechanism by which effciency of oxidative phosphorylation becomes altered. Metabolic-associated fatty liver disease (MAFLD) is a progressive disorder characterized by oxidative stress and accumulation of fat in hepatocytes that can lead to metabolic dysfunction-associated steatohepatitis (MASH). We hypothesized that MAFLD alters mitochondrial membrane lipids to induce mitochondrial electron transfer ineffciency and promotes oxidative stress. Lipid mass spectrometric analyses in four models of MAFLD (high-fat diet, ob/ob, carbon tetrachloride administration (CCI4), and Gubra-Amylin-NASH® diet), revealed that low mitochondrial cardiolipin (CL) is a common signature associated with all MAFLD models. In hepa1-6 cells, lentivirus-mediated knockdown of cardiolipin synthase (CLS, generates CL in mitochondria) was suffcient to reduce respiration and increase electron leak. Liver mitochondria from mice with a hepatocyte-specific knockout of CLS exhibited defects in mitochondrial bioenergetics including increased electron leak, concomitant to pathogenesis of MASH. Together, these findings provide evidence that a reduction in hepatic cardiolipin may drive the progression of MAFLD to induce MASH pathology. R01GM144613-03R01AG074535-02S2R01DK107397 07. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Tissue-specific differences in Ca2+ sensitivity of the mitochondrial permeability transition pore (PTP). Experiments in male rat liver and heart.

Permeability transition pore (PTP) opening dissipates ion and electron gradients across the internal mitochondrial membrane (IMM), including excess Ca2+ in the mitochondrial matrix. After opening, immediate PTP closure must follow to prevent outer membrane disruption, loss of cytochrome c, and eventual apoptosis. Flickering, defined as the rapid alternative opening/closing of PTP, has been reported in heart, which undergoes frequent, large variations in Ca2+. In contrast, in tissues that undergo depolarization events less often, such as the liver, PTP would not need to be as dynamic and thus these tissues would not be as resistant to stress. To evaluate this idea, it was decided to follow the reversibility of the permeability transition (PT) in isolated murine mitochondria from two different tissues: the very dynamic heart, and the liver, which suffers depolarizations less frequently. It was observed that in heart mitochondria PT remained reversible for longer periods and at higher Ca2+ loads than in liver mitochondria. In all cases, Ca2+ uptake was inhibited by ruthenium red and PT was delayed by Cyclosporine A. Characterization of this phenomenon included measuring the rate of oxygen consumption, organelle swelling and Ca2+ uptake and retention. Results strongly suggest that there are tissue-specific differences in PTP physiology, as it resists many more Ca2+ additions before opening in a highly active organ such as the heart than in an organ that seldom suffers Ca2+ loading, such as the liver.

In vivo chronic exposure to inorganic mercury worsens hypercholesterolemia, oxidative stress and atherosclerosis in the LDL receptor knockout mice

Heavy metal exposure leads to multiple system dysfunctions. The mechanisms are likely multifactorial and involve inflammation and oxidative stress. The aim of this study was to evaluate markers and risk factors for atherosclerosis in the LDL receptor knockout mouse model chronically exposed to inorganic mercury (Hg) in the drinking water. Results revealed that Hg exposed mice present increased plasma levels of cholesterol, without alterations in glucose. As a major source and target of oxidants, we evaluated mitochondrial function. We found that liver mitochondria from Hg treated mice show worse respiratory control, lower oxidative phosphorylation efficiency and increased H2O2 release. In addition, Hg induced mitochondrial membrane permeability transition. Erythrocytes from Hg treated mice showed a 50% reduction in their ability to take up oxygen, lower levels of reduced glutathione (GSH) and of antioxidant enzymes (SOD, catalase and GPx). The Hg treatment disturbed immune system cells counting and function. While lymphocytes were reduced, monocytes, eosinophils and neutrophils were increased. Peritoneal macrophages from Hg treated mice showed increased phagocytic activity. Hg exposed mice tissues present metal impregnation and parenchymal architecture alterations. In agreement, increased systemic markers of liver and kidney dysfunction were observed. Plasma, liver and kidney oxidative damage indicators (MDA and carbonyl) were increased while GSH and thiol groups were diminished by Hg exposure. Importantly, atherosclerotic lesion size in the aorta root of Hg exposed mice were larger than in controls. In conclusion, in vivo chronic exposure to Hg worsens the hypercholesterolemia, impairs mitochondrial bioenergetics and redox function, alters immune cells profile and function, causes several tissues oxidative damage and accelerates atherosclerosis development.

Fatty acid oxidation drives mitochondrial hydrogen peroxide production by α-ketoglutarate dehydrogenase

In the present study, we examined the mitochondrial hydrogen peroxide (mH2O2) generating capacity of α-ketoglutarate dehydrogenase (KGDH) and compared it to components of the electron transport chain using liver mitochondria isolated from male and female C57BL6N mice. We show for the first time there are some sex dimorphisms in the production of mH2O2 by electron transport chain complexes I and III when mitochondria are fueled with different substrates. However, in our investigations into these sex effects, we made the unexpected and compelling discovery that 1) KGDH serves as a major mH2O2 supplier in male and female liver mitochondria and 2) KGDH can form mH2O2 when liver mitochondria are energized with fatty acids but only when malate is used to prime the Krebs cycle. Surprisingly, 2-keto-3-methylvaleric acid (KMV), a site-specific inhibitor for KGDH, nearly abolished mH2O2 generation in both male and female liver mitochondria oxidizing palmitoyl-carnitine. KMV inhibited mH2O2 production in liver mitochondria from male and female mice oxidizing myristoyl-, octanoyl-, or butyryl-carnitine as well. S1QEL 1.1 (S1) and S3QEL 2 (S3), compounds that inhibit reactive oxygen species generation by complexes I and III, respectively, without interfering with OxPhos and respiration, had a negligible effect on the rate of mH2O2 production when pyruvate or acyl-carnitines were used as fuels. However, inclusion of KMV in reaction mixtures containing S1 and/or S3 almost abolished mH2O2 generation. Together, our findings suggest KGDH is the main mH2O2 generator in liver mitochondria, even when fatty acids are used as fuel.

The striking differences in the bioenergetics of brain and liver mitochondria are enhanced in mitochondrial disease

Mitochondrial disorders are hallmarked by the dysfunction of oxidative phosphorylation (OXPHOS) yet are highly heterogeneous at the clinical and genetic levels. Striking tissue-specific pathological manifestations are a poorly understood feature of these conditions, even if the disease-causing genes are ubiquitously expressed. To investigate the functional basis of this phenomenon, we analyzed several OXPHOS-related bioenergetic parameters, including oxygen consumption rates, electron transfer system (ETS)-related coenzyme Q (mtCoQ) redox state and production of reactive oxygen species (ROS) in mouse brain and liver mitochondria fueled by different substrates. In addition, we determined how these functional parameters are affected by ETS impairment in a tissue-specific manner using pathologically relevant mouse models lacking either Ndufs4 or Ttc19, leading to Complex I (CI) or Complex III (CIII) deficiency, respectively. Detailed OXPHOS analysis revealed striking differences between brain and liver mitochondria in the capacity of the different metabolic substrates to fuel the ETS, reduce the ETS-related mtCoQ, and to induce ROS production. In addition, ETS deficiency due to either CI or CIII dysfunction had a much greater impact on the intrinsic bioenergetic parameters of brain compared with liver mitochondria. These findings are discussed in terms of the still rather mysterious tissue-specific manifestations of mitochondrial disease.

Ingestion of triglycerides containing medium- and long-chain fatty acids can increase metabolism of ingested long-chain triglycerides in overweight persons.

Medium-chain fatty acids (MCFAs) have attracted considerable attention for preventing or improving obesity, which is a recognized risk factor for lifestyle-related diseases. Medium- and long-chain triglycerides (MLCTs) are expected to improve the metabolism of ingested long-chain triglycerides (LCTs). However, previous studies have reported mixed results. In this study, the effect of ingestion of MLCTs was evaluated on the metabolism of LCTs and compared to the ingestion of rapeseed oil (control oil). A randomized, double-blind, placebo-controlled crossover study was performed among sedentary participants with BMIs ranging from 25 below 30 kg/m2. Thirty participants were asked to ingest either 14 g of MLCTs or a control oil for 4 weeks. The metabolism of ingested LCTs was evaluated by measuring isotopically labeled carbon dioxide released by the degradation of carbon-13 (13C)-labeled LCTs. Ingestion of MLCTs markedly enhanced the metabolism of ingested LCTs by comparison to the control oil. The findings of this study suggest that ingestion of MLCTs may enhance the metabolism of dietary LCTs through activation of β-oxidation in liver mitochondria, which may increase the metabolic kinetics of ingested long-chain fatty acid (LCFAs). https://center6.umin.ac.jp/cgi-open-bin/ctr/ctr_view.cgi?recptno=R000053101, identifier: UMIN000046604.

Reversal of mitochondrial permeability transition pore and pancreas degeneration by chloroform fraction of Ocimum gratissimum (L.) leaf extract in type 2 diabetic rat model.

Introduction: Unmanaged Diabetes Mellitus (DM) usually results to tissue wastage because of mitochondrial dysfunction. Adverse effects of some drugs used in the management of DM necessitates the search for alternative therapy from plant origin with less or no side effects. Ocimum gratissimum (L.) (OG) has been folklorically used in the management of DM. However, the mechanism used by this plant is not fully understood. This study was designed to investigate the effects of chloroform fraction of OG leaf (CFOG) in the reversal of tissue wastage in DM via inhibition of mitochondrial-mediated cell death in streptozotocin (STZ)-induced diabetic male Wistar rats. Methods: Air-dried OG leaves were extracted with methanol and partitioned successively between n-hexane, chloroform, ethylacetate and methanol to obtain their fractions while CFOG was further used because of its activity. Diabetes was induced in fifteen male Wistar rats, previously fed with high fat diet (28days), via a single intraperitoneal administration of STZ (35mg/kg). Diabetes was confirmed after 72h. Another five fed rats were used as the normal control, treated with corn oil (group 1). The diabetic animals were grouped (n = 5) and treated for 28days as follows: group 2 (diabetic control: DC) received corn oil (10mL/kg), groups 3 and 4 were administered 400mg/kg CFOG and 5mg/kg glibenclamide, respectively. Body weight and Fasting Blood Glucose (FBG) were determined while Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) and beta cell (HOMA-β), and pancreatic tissue regenerating potential by CFOG were assessed. Activity-guided purification and characterization of the most active principle in CFOG was done using chromatographic and NMR techniques. The animals were sacrificed after 28days, blood samples were collected and serum was obtained. Liver mitochondria were isolated and mitochondrial permeability transition (mPT) was investigated by spectrophotometry. Results: CFOG reversed diabetic-induced mPT pore opening, inhibited ATPase activity and lipid peroxidation. CFOG reduced HOMA-IR but enhanced HOMA-β and caused regeneration of pancreatic cells relative to DC. Lupanol was a major metabolite of CFOG. Discussion: Normoglycemic effect of CFOG, coupled with reversal of mPT, reduced HOMA-IR and improved HOMA-β showed the probable antidiabetic mechanism and tissue regenerating potentials of OG.

The Role of Epigenetic Control of Mitochondrial (Dys)Function in MASLD Onset and Progression.

Metabolic dysfunction-associated steatotic fatty liver disease (MASLD), a novel definition for NAFLD, represents one of the most common causes of liver disease, and its incidence is increasing worldwide. It is characterized by a complex etiopathogenesis in which mitochondrial dysfunction exerts a pivotal role together with alteration of lipid metabolism, inflammation, and oxidative stress. Nutrients and bioactive compounds can influence such mechanisms so that changes in diet and lifestyle are regarded as important treatment strategies. Notably, natural compounds can exert their influence through changes of the epigenetic landscape, overall resulting in rewiring of molecular networks involved in cell and tissue homeostasis. Considering such information, the present review aims at providing evidence of epigenetic modifications occurring at mitochondria in response to natural and bioactive compounds in the context of liver (dys)function. For this purpose, recent studies reporting effects of compounds on mitochondria in the context of NAFLD/MASLD, as well as research showing alteration of DNA methylation and non-coding RNAs-related circuits occurring at liver mitochondria, will be illustrated. Overall, the present review will highlight the importance of understanding the bioactive compounds-dependent epigenetic modulation of mitochondria for improving the knowledge of MASLD and identifying biomarkers to be employed for effective preventative strategies or treatment protocols.

Blockade of L-type calcium channels alters hepatic mitochondrial function in insulin-resistant rats

Mitochondria are central organelles in maintaining energy and oxidative homeostasis. Despite intensive research, the function of mitochondria and the mechanisms of its regulation under physiological conditions and in insulin resistance require detailed investigation. The aim of this study was to investigate the effect of blockade L-type calcium channels in insulin-resistant rats on hepatic mitochondrial oxidative function and changes in its oxidative status. Insulin resistance was modeled in 6-month-old male Wistar rats by 14 days of high-fat feeding. Standard fed animals served as controls. Verapamil was administered intraperitoneally at a dose of 1 mg/kg to block L-type calcium channels. Indicators of pro- and antioxidant systems (active products of tiobarbituric acid, reduced glutathione, catalase, Cu,Zn-superoxide dismutase) were assayed in the liver homogenate extracted from anesthetized animals after 3 h. Mitochondrial function was studied by the Chance polarographic method using different metabolic substrates. It was shown that in intact animals blockade of L-type calcium channels reduced the efficiency of mitochondrial respiration (V3/V4) in liver mitochondria during oxidation of all substrates through an inhibitory effect on the phosphorylation respiration (V3) and a stimulatory effect on the controlled respiration (V4), without affecting the oxidative status of the liver. In rats with insulin resistance the rate of V3 during oxidation of both NAD- and FADdependent substrates was decreased, violations of oxidative status and increased antioxidant protection were detected. However, in insulin-resistant rats blockade of L-type calcium channels significantly enhanced basic respiration (V2) during NAD-dependent substrate oxidation, V3 and V4 during palmitoyl lipid substrate oxidation, reduced the V3/V4 ratio compared to control, and partially or fully restored the violation of the oxidative status. This may indicate the involvement of calcium mechanisms in the disturbance of the oxidative status of the liver and the regulation of energy metabolism in mitochondria during insulin resistance.

Bioenergetic functions of mitochondria in liver, pancreatic acinar cells, and sperm cells of rats fed short-term high-fat or high-fat high-sugar diets

An unhealthy diet often is a cause of obesity, chronic inflammation, and metabolic disruption in multiple organs. However, the direct influence of elevated lipid or sugar consumption on liver, pancreatic, and sperm mitochondria is not well understood. The aim of the study was to investigate the functional activity of mitochondria of liver, pancreatic acinar cells, and sperm cells in rats on a short-term (7 weeks) diet with high fat or high fat and high sugar content. Male Wistar rats were on a basic, high-fat or high-fat high-sugar diet for 7 weeks. At the end of the experiment, visceral fat mass, blood glucose and lipids were measured. Mitochondrial functional activity was evaluated with oxygen consumption assay. In isolated pancreatic acinar cells, NAD(P)H autofluorescence and mitochondrial membrane potential were also studied. No difference in body mass was observed between the 3 groups at the end of the experiment. Visceral fat mass was slightly but significantly elevated in rats on a high-fat high-sugar diet. Both diets did not affect plasma glucose or triglyceride levels but caused a modest elevation of total plasma cholesterol. Respiration and oxidative phosphorylation of isolated liver mitochondria were not affected by any experimental diet. In pancreatic acinar cells, a high-fat diet caused a significant decrease of basal respiration by ~15%, but no effects were observed on the maximal rate of uncoupled respiration, mitochondrial membrane potential, or NAD(P)H autofluorescence. In these cells, a ketone body 3-hydroxybutyrate caused elevation of uncoupled respiration and NAD(P)H level irrespectively of the diet. Diets did not cause any change in sperm concentration, viability or motility. Surprisingly, in animals on a high-fat high-sugar diet, a significant increase in both basal and maximal respiration of sperm cells was observed. Collectively, these data show that while the elevated fat and sugar content in the diet does not cause significant obesity, no detrimental effects on mitochondria of the liver, pancreas, and sperm cells are observed. Keywords: diet, liver, mitochondria, pancreatic acinar cells, sperm

Ketone Bodies in Diabetes Mellitus: Friend or Foe?

In glucose-deprived conditions, ketone bodies are produced by the liver mitochondria, through the catabolism of fatty acids, and are used peripherally, as an alternative energy source. Ketones are produced in the body under normal conditions, including during pregnancy and the neonatal period, when following a ketogenic diet (KD), fasting, or exercising. Additionally, ketone synthesis is also augmented under pathological conditions, including cases of diabetic ketoacidosis (DKA), alcoholism, and several metabolic disorders. Nonetheless, diet is the main regulator of total body ketone concentrations. The KDs are mimicking the fasting state, altering the default metabolism towards the use of ketones as the primary fuel source. Recently, KD has gained recognition as a medical nutrition therapy for a plethora of metabolic conditions, including obesity and diabetes mellitus (DM). The present review aims to discuss the role of ketones, KDs, ketonemia, and ketonuria in DM, presenting all the available new evidence in a comprehensive manner.

Long term survival and abnormal liver fat accumulation in mice with specific thymidine kinase 2 deficiency in liver tissue.

Deficiency in thymidine kinase 2 (TK2) causes mitochondrial DNA depletion. Liver mitochondria are severely affected in Tk2 complete knockout models and have been suggested to play a role in the pathogenesis of the Tk2 knockout phenotype, characterized by loss of hypodermal fat tissue, growth retardation and reduced life span. Here we report a liver specific Tk2 knockout (KO) model to further study mechanisms contributing to the phenotypic changes associated with Tk2 deficiency. Interestingly, the liver specific Tk2 KO mice had a normal life span despite a much lower mtDNA level in liver tissue. Mitochondrial DNA encoded peptide COXI did not differ between the Tk2 KO and control mice. However, the relative liver weight was significantly increased in the male Tk2 KO mouse model. Histology analysis indicated an increased lipid accumulation. We conclude that other enzyme activities can partly compensate Tk2 deficiency to maintain mtDNA at a low but stable level throughout the life span of the liver specific Tk2 KO mice. The lower level of mtDNA was sufficient for survival but led to an abnormal lipid accumulation in liver tissue.

State of peroxidation in the brain and liver in experimental diabetes and its correction possibility with niacin-oxyethylendiphosphonatogermanate

The development of pharmacological methods to control oxidant stress manifestations in diabetes can be achieved using organic complex germanium compounds. One of the promising compounds is niacin-oxyethylenediphosphonategermanate (MIGU-4), which is an effective corrector of lipid metabolism and a stabilizer of the lipid layer of erythrocyte and hepatocyte membranes in the streptozotocin-induced diabetic model. The aim of the work is to determine the dynamics of the malondialdehyde (MDA) content, the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPO) in the brain tissue, as well as the diene conjugate (DC) content, lipid hydroperoxides (LHP), reduced glutathione (GSH) in the liver mitochondria, the serum activities of aspartate and alanine aminotransferases (AST and ALT) in experimental diabetes mellitus with the correction using a complex compound of germanium with nicotinic acid – MIGU-4 and insulin, insulin alone as well as in comparison with effectiveness of vitamin E supplementation. Materials and methods. Diabetes was induced in male Wistar rats by intraperitoneal injection of streptozotocin (60.0 mg/kg). MIGU-4 was administered intraperitoneally at a dose of ED50 that was 25.0 mg/kg. Mitochondrial membranes were obtained by differential centrifugation of the liver tissue. In rats with confirmed diabetes, parameters of peroxidation and antioxidant protection were studied by generally accepted biochemical and biophysical methods. Results. Glucose levels were 29.4 % lower in rats treated with MIGU-4 (25.0 mg/kg) after 8 weeks of streptozotocin-induced diabetes than those in untreated diabetic rats (p < 0.05), while a decrease was 39.0 % (p < 0.05) upon insulin treatment, and 47.2 % (p < 0.05) – with combined use of the drugs. The MDA content in the brain tissue was 3.48 times higher than that in the control (p < 0.05). At the same time, the activities of SOD and CAT were decreased by 46.4 % and 32.0 %, respectively, the activity of GPO was decreased by half (p < 0.05). In the liver mitochondria, the DC content exceeded that in the control by 53.5 % (p < 0.05), and the MDA and LHP levels were 2.48 times and 31.7 % higher (p < 0.05), respectively. AST activity was almost doubled, ALT activity was 5.48 times increased. Insulin and the biologically active substance MIGU-4 used alone exerted a moderate corrective effect. The combined use of the drugs caused definite therapeutic and preventive effects as the brain content of MDA was 2.4 times decreased, the activity of SOD, CAT and GPO was increased by 45.6 %, 35.2 % and 67.3 % (p < 0.05), respectively. In the liver mitochondria, the DC, MDA, and LHP contents were decreased by 41.0 %, 53.3 %, and 28.4 %, respectively, compared to those in rats with diabetes (p < 0.05). The activity of SOD and CAT as well as GSH content were increased by 2.7 times, 51.9 %, and 23.0 % (p < 0.05), respectively. AST and ALT activities were 42.2 % and 74.3 % (p < 0.05) reduced, respectively. The combined use of MIGU-4 (25.0 mg/kg) and vitamin E (250.0 mg/kg) caused a decrease in the brain MDA content by 70.1 % (p < 0.05) in experimental rats. The combination of MIGU-4 and vitamin E increased the activity of GPO by 58.3 % (p < 0.05) as compared to rats with diabetes, indicating cumulative effects of these drugs. Conclusions. The use of MIGU-4 prevents the occurrence of oxidant stress manifestations in the brain tissue and liver mitochondria in streptozotocin-induced diabetes. The corrective effect size of MIGU-4 (25.0 mg/kg) corresponds to that of vitamin E (250.0 mg/kg).

Sirtuin 5 Alleviates Liver Ischemia/Reperfusion Injury by Regulating Mitochondrial Succinylation and Oxidative Stress.

Aims: Mitochondrial dysfunction is the primary mechanism of liver ischemia/reperfusion (I/R) injury. The lysine desuccinylase sirtuin 5 (SIRT5) is a global regulator of the mitochondrial succinylome and has pivotal roles in mitochondrial metabolism and function; however, its hepatoprotective capacity in liver I/R remains unclear. In this study, we established liver I/R model in SIRT5-silenced and SIRT5-overexpressed mice to examine the role and precise mechanisms of SIRT5 in liver I/R injury. Results: Succinylation was strongly enriched in liver mitochondria during I/R, and inhibiting mitochondrial succinylation significantly attenuated liver I/R injury. Importantly, the levels of the desuccinylase SIRT5 were notably decreased in liver transplant patients, as well as in mice subjected to I/R and in AML12 cells exposed to hypoxia/reoxygenation. Furthermore, SIRT5 significantly ameliorated liver I/R-induced oxidative injury, apoptosis, and inflammation by regulating mitochondrial oxidative stress and function. Intriguingly, the hepatoprotective effect of SIRT5 was mediated by PRDX3. Mechanistically, SIRT5 specifically desuccinylated PRDX3 at the K84 site, which enabled PRDX3 to alleviate mitochondrial oxidative stress during liver I/R. Innovation: This study denoted the new effect and mechanism of SIRT5 in regulating mitochondrial oxidative stress through lysine desuccinylation, thus preventing liver I/R injury. Conclusions: Our findings demonstrate for the first time that SIRT5 is a key mediator of liver I/R that regulates mitochondrial oxidative stress through the desuccinylation of PRDX3, which provides a novel strategy to prevent liver I/R injury. Antioxid. Redox Signal. 40, 616-631.