PM2.5 toxicity in blood impairs cardiac redox balance and promotes mitochondrial dysfunction in rat heart that further aggravates ischemia reperfusion injury by modulating PI3K/AKT/mTOR/NF-kB signaling axis.

According to the pathophysiological mechanisms linking particulate matter (PM2.5) exposure and cardiovascular diseases, PM2.5 may directly translocate into the blood stream and remote target organs and thereby induce cardiovascular effects. The toxicity of PM2.5 is known to induce oxidative stress in pulmonary tissue, but its impact on the redox state in heart (distant organ) is unknown and how it modulates the cardiac response to ischemia reperfusion (IR) remains unclear. In the present study, we evaluated the toxic effect of PM2.5 on cardiac physiology in the presence and absence of IR after introducing PM2.5 into the blood. Female Wistar rats were injected with diesel particulate matter (DPM) via i.p & i.v routes at a concentration of 10 µg/ml. The toxic impact of PM2.5 not only adversely affects the cardiac ultra-structure (leading to nuclear infiltration, edema, irregularities in heart muscle and nuclear infiltration), but also altered the cellular redox balance, elevated inflammation and promoted the upregulation of proapoptotic mediator genes at the basal level of myocardium. The results showed alterations in cardiac ultrastructure, elevated oxidative stress and significant redox imbalance, increased inflammation and proapoptotic mediators at the basal level of myocardium. Moreover, the cardioprotective pro survival signaling axis was declined along with an increased NF-kB activation at the basal level. IR inflicted further injury with deterioration of cardiac hemodynamic indices (Heart rate [HR], Left ventricular developed pressure [LVDP], Left ventricular end-diastolic pressure [LVEDP] and rate pressure product [RPP]) along with prominent inactivation of signaling pathways. Furthermore, the levels of GSH/GSSG, NADH/NAD, NADPH/NADP were significantly low along with increased lipid peroxidation in mitochondria of PM2.5 treated IR rat hearts. This observation was supported by downregulation of glutaredoxin and peroxiredoxin genes in the myocardium. Similarly the presence of oxidative stress inducing metals was found at a higher concentration in cardiac mitochondria. Thus, the toxic impact of PM2.5 in heart augment the IR associated pathological changes by altering the physiological response, initiating cellular metabolic alterations in mitochondria and modifying the signaling molecules.

High-fat diet-induced mitochondrial dysfunction is associated with loss of protection from ischemic preconditioning in renal ischemia reperfusion.

Consumption of high-fat diet (HFD) promotes mitochondrial dysfunction and the latter act as a critical factor in determining the severity of ischemia-reperfusion (IR) injury in different cell types. Ischemic preconditioning (IPC), a well-known protocol that render IR protection in kidney works via mitochondria. In the present study, we evaluated how HFD kidney with underlying mitochondrial changes respond to precondition protocol after IR induction. Wistar male rats were used in this study and were divided into two groups: SD (standard diet; n = 18) and HFD (high-fat diet; n = 18), which were further subdivided into sham, ischemia-reperfusion, and precondition groups at the end of the dietary regimen. Blood biochemistry, renal injury marker, creatinine clearance (CrCl), mitochondrial quality (fission, fusion, and phagy), mitochondrial function via ETC enzyme activities and respiration, and signalling pathway were analysed. Sixteen weeks of HFD administration to the rat deteriorated the renal mitochondrial health measured via 10% decline in mitochondrial respiration index ADP/O (in GM), reduced mitochondrial copy number (55%), biogenesis (56%), low bioenergetics potential (19% complex I + III and 15% complex II + III), increased oxidative stress, and reduced expression of mitochondrial fusion genes compared with SD rats. IR procedure in HFD rat kidney inflicted significant mitochondrial dysfunction and further deteriorated copy number along with impaired mitophagy and mitochondrial dynamics. IPC could effectively ameliorate the renal ischemia injury in normal rat but failed to provide similar kind of protection in HFD rat kidney. Even though the IR-associated mitochondrial dysfunction in both normal and HFD rats were similar, the magnitude of overall dysfunction and corresponding renal injury and compromised physiology was high in HFD rats. This observation was further confirmed via in vitro protein translation assay in isolated mitochondria from normal and HFD rat kidney that showed significantly reduction in the response ability of mitochondria in HFD. In conclusion, the deteriorated mitochondrial function and its quality along with low mitochondrial copy number and downregulation of mitochondrial dynamic gene exhibited by HFD rat kidney augments the sensitivity of renal tissue towards the IR injury which leads to the compromised protective ability by ischemic preconditioning.

DNA hypomethylation by fisetin preserves mitochondria functional genes and contributes to the protection of I/R rat heart.

Myocardial I/R can alter the expression of different sets of cardiac genes that negatively influence the I/R outcome via epigenetic modifications. Fisetin is known to be cardioprotective against I/R, but its underlying epigenetic mode of action is not known and is addressed in the present study. Male Wistar rats were subjected to I/R by using the Langendorff perfusion system. Fisetin (20 mg/kg; i.p.) was administered before I/R induction, followed by the measurement of cardiac injury, hemodynamics, physiological indices, the differential expression of genes that regulate DNA methylation, and the function of mitochondria were performed. Fisetin administered I/R rat heart significantly reduced the global DNA hypermethylation and infarct size with an improved physiological recovery, measured via RPP (81%) and LVDP (82%) from the I/R control. Additionally, we noted decreased expression of the DNMT1 gene by 35% and increased expression of the TET1, TET2, and TET3 genes in fisetin-treated I/R rat hearts. Molecular docking analysis data reveals that the fisetin inhibits DNMT1 at the substrate binding site with minimum binding energy (- 8.2 kcal/mol) compared to the DNMT1 inhibitor, 5-azacytidine. Moreover, fisetin-treated I/R heart reversed the expression of the I/R-linked declined expression of bioenergetics genes (MT-ND1, MT-ND2, MT-ND4, MT-Cyt B, MT-COX1, MT-COX2, MT-ATP6), mitochondrial fission gene (Fis1), replication control genes PGC-1α, POLG, and TFAM to near-normal level. Based on the above findings, we demonstrated that fisetin possesses the ability to modulate the expression of different mitochondrial genes via influencing the global DNA methylation in cardiac tissue, which contributes significantly to the improved contractile function and thereby renders cardioprotection against I/R.

Impaired renal ischemia reperfusion recovery after bilateral renal artery ligation in rats treated with adenine: role of renal mitochondria.

Vascular calcification (VC) and ischemia reperfusion (IR) injury is characterised to have mitochondrial dysfunction. However, the impact of dysfunctional mitochondria associated with vascular calcified rat kidney challenged to IR is not explored and is addressed in the present study. Male Wistar rats were treated with adenine for 20 days to induce chronic kidney dysfunction and VC. After 63 days, renal IR protocol was performed with subsequent recovery for 24 h and 7 days. Various mitochondrial parameters and biochemical assays were performed to assess kidney function, IR injury and its recovery. Adenine-induced rats with VC, decreased creatinine clearance (CrCl), and severe tissue injury demonstrated an increase in renal tissue damage and decreased CrCl after 24 h of IR (CrCl in ml: IR-0.220.02, VC-IR-0.050.01). Incidentally, the 24 h IR pathology in kidney was similar in both VC-IR and normal rat IR. But, the magnitude of dysfunction was higher with VC-IR due to pre-existing basal tissue alterations. We found severed deterioration in mitochondrial quantity and quality supported by low bioenergetic function in both VC basal tissue and IR challenged sample. However, post 7 days of IR, unlike normal rat IR, VC rat IR did not improve CrCl and corresponding mitochondrial damage in terms of quantity and its function were observed. Based on the above findings, we conclude that IR in VC rat adversely affect the post-surgical recovery, mainly due to the ineffective renal mitochondrial functional restoration from the surgery.

PM2.5 from diesel exhaust attenuated fisetin mediated cytoprotection in H9c2 cardiomyocytes subjected to ischemia reoxygenation by inducing mitotoxicity.

The impact of PM2.5 from diesel exhaust (termed as diesel particulate matter (DPM)) on ischemia re-oxygenation (IR) injury and the consequent effect of fisetin to attenuate this injury remains unclear. IR was induced in H9c2 cells after 24 hrs of fisetin treatment. The cells when incubated with 100 µg/mL of DPM followed by IR, induced 60% cell death which was escalated to 78% with DPM exposure. Fisetin significantly attenuated IR induced cytotoxicity, improved mitochondrial activity and reduced oxidative stress in normal cells but failed to render protection against IR in presence of DPM. Isolated mitochondria experiment confirmed the mitotoxic effect of DPM. Immunoblot analysis established the failure of fisetin to activate PI3K/Akt signaling pathway. Based on the above observations, we concluded that fisetin mediated protection against IR was abrogated with DPM exposure due to augmented mitochondrial dysfunction and inactivation of PI3K/Akt signaling pathway.

Hydrogen sulfide postconditioning rendered cardioprotection against myocardial ischemia-reperfusion injury is compromised in rats with diabetic cardiomyopathy.

The present study aimed to investigate the efficacy of hydrogen sulfide (H2S) post-conditioning (HPOC) against ischemia-reperfusion (I/R) challenged diabetic rat hearts with or without cardiomyopathy using the Langendorff perfusion system. Male Wistar rats were randomly divided into different groups such as normal, diabetes mellitus (DM), and diabetic cardiomyopathy (DCM). Hearts from these groups were subjected to normal perfusion, I/R, and HPOC and were analyzed for cardiac physiology, cardiomyocyte injury, mitochondrial function, oxidative stress, and H2S metabolism. The results showed that HPOC protocol reduced the cardiac injury and improved the haemodynamics in normal and DM effectively, but not in DCM (RPP in mmHg*beats/min*103: HPOC- 32 ± 2, DM-HPOC-19 ± 1, DCM-HPOC-6 ± 2, LVDP in mmHg: HPOC- 96 ± 3, DM-HPOC-73 ± 2, DCM-HPOC-50 ± 3). DCM rats at the basal level exhibited perturbed myocardial architecture, mitochondrial dysfunction, and impaired glycolytic flux that failed to improve by HPOC treatment after I/R. HPOC exhibited a nominal improvement in the gene expression and activities of the H2S metabolizing enzymes such as cystathionine beta-synthase, rhodanese, and cystathionine-gamma-lyase in DCM hearts. Collectively, our results suggest that altered myocardial architecture along with exacerbated oxidative stress and mitochondrial dysfunction contribute towards the failure of HPOC cardioprotection against I/R-induced myocardial tissue injury in DCM.

Inhalation of PM2.5 from diesel exhaust promote impairment of mitochondrial bioenergetics and dysregulate mitochondrial quality in rat heart: implications in isoproterenol-induced myocardial infarction model.

Aim: Ambient exposure of PM2.5 from diesel exhaust (termed as diesel particulate matter [DPM]) can induce cardiotoxicity that can be manifested into myocardial ischemia/infarction, where the survival depends on mitochondrial function. The mechanism for DPM-induced mitochondrial dysfunction is yet to be elucidated and the consequential impact of impaired mitochondria on the severity of myocardial infarction (MI) has not been established.Materials and methods: Female Wistar rats were exposed to DPM (0.5 mg/ml) for 3 h daily (to achieve a PM2.5 concentration of 250 µg/m3) for 21 d trailed by an induction of MI using isoproterenol (ISO).Conclusion: DPM exposure altered the basal ECG pattern and increased heart weight (HW) to body weight (BW) ratio from control. Loss of mitochondrial quality in the cardiac tissue was observed in DPM exposed animals, measured via declined ETC enzyme activity, reduced ATP levels, high oxidative stress, low mitochondrial copy number, and low expression of the mitochondrial genes involved in mitophagy (PINK and PARKIN) and mitochondrial fusion (MFN-1). Subsequent induction of MI in DPM exposed animals (DPM + ISO) further deteriorated the normal sinus rhythm, accompanied by elevated plasma CK and LDH level, increased myocardial caspase activity, downregulation of Peroxisome proliferator-activated receptor-gamma coactivator (PGC1-α), transcription factor A (TFAM), DNA polymerase subunit gamma (POLG), and other mitochondrial quality control genes. Based on these results, we conclude that DPM alters the electrophysiology and ultrastructure of the heart that aggravates the MI-induced cardiotoxicity, where the diminished mitochondrial quality can be the potential contributor.

Resveratrol-mediated cardioprotection against myocardial ischemia-reperfusion injury was revoked by statin-induced mitochondrial alterations.

Resveratrol is well known for its antioxidant potential and ability to preserve mitochondrial function, reported attenuating ischemia-reperfusion (IR) injury in the heart. The present study investigates resveratrol on IR injury in rat hearts treated with statin for 14 days. Male Wistar rats were used in this study, and statin-induced cardiac metabolic alterations were monitored after the administration of simvastatin (80 mg/kg). IR was instigated by the Langendroff perfusion system and measured the physiological and biochemical changes. The basal level changes in ECG, ANP, and BNP expression and CoenzymeQ10 level were altered in statin-treated animals compared to the normal rat heart. The animals treated with statin demonstrated higher IR injury (measured via low rate pressure product (88.4%), increased histological alterations, prominent mitochondrial dysfunction (NQR: IR-72%, Stat IR-67%; SQR: IR-71%, Stat IR-74%; COX: IR-58%, Stat IR-52%) than the normal rat heart underwent similar protocols. Administration of heart with resveratrol recovered the IR associated hemodynamic indices in normal heart subjected to IR but failed to impart a similar effect in the statin-treated heart. Our results demonstrated that resveratrol failed to reverse the IR-associated cardiac injury and functional abnormalities in statin-treated rat hearts subjected to IR but effective in IR challenged normal heart.

Preconditioning the rat heart with 5-azacytidine attenuates myocardial ischemia/reperfusion injury via PI3K/GSK3ß and mitochondrial KATP signaling axis.

5-Azacytidine is well known for its clinical usage in cancer treatments. The present study investigates the role of 5-azacytidine as a cardioprotective agent to ameliorate ischemia/reperfusion (I/R) injury. The cardioprotective effect of 5-azacytidine was evaluated in three experimental models: in vitro, ex vivo, and in vivo. The cardioprotective effect was evaluated via cell viability, hemodynamic indices, infarct size measurement, and assessment of histopathology, oxidative stress, and mitochondrial function. The experiments were repeated in the presence of PI3K/GSK3ß and mitochondrial KATP (mtKATP ) cardioprotective signaling pathway inhibitors to understand the underlying mechanism. 5-Azacytidine improved the cell viability by 29% in I/R-challenged H9C2 cells. Both isolated rat heart and LAD ligation model confirmed the infarct sparing effect of 5-azacytidine against I/R. It also provided a beneficial effect by normalizing the altered hemodynamics, reducing the infarct size and cardiac injury markers, reversing the perturbation of mitochondria, reduced oxidative stress, and improved the pPI3K and pAKT protein expression from I/R. In addition, it also augmented the activation of PI3K/AKT and mtKATP signaling pathway, confirmed by using wortmannin (PI3K inhibitor), SB216763 (GSK3ß inhibitor), and glibenclamide (mtKATP channel closer). The effectiveness of 5-azacytidine as a cardioprotective agent is attributed to its activation of the PI3K/GSK3ß and mtKATP channel signaling axis, thereby preserving mitochondrial function and reducing oxidative stress.

Diabetic animal fed with high-fat diet prevents the protective effect of myocardial ischemic preconditioning effect in isolated rat heart perfusion model.

Diabetic heart (diabetes mellitus [DM]) has been shown to attenuate the beneficial effect of ischemic preconditioning (IPC) in rat heart. But the effect of IPC on diabetic rat heart that develops myopathy remains unclear. This study was designed to test the impact of IPC on diabetic cardiomyopathy (DCM) rat heart. Male Wistar rats were grouped as (a) normal, (b) DM (streptozotocin: 65 mg/kg; fed with normal diet), and (c) DCM (streptozotocin: 65 mg/kg; fed with high-fat diet). Isolated rat hearts from each group were randomly subjected to (a) normal perfusion, (b) ischemia-reperfusion (I/R), and (c) IPC procedure. At the end of the perfusion experiments, hearts were analyzed for injury, contractile function, mitochondrial activity, and oxidative stress. The results obtained from hemodynamics, cardiac injury markers, and caspase-3 activity showed that DCM rat displayed prominent I/R-associated cardiac abnormalities than DM rat heart. But the deteriorated physiological performance and cardiac injury were not recovered in both DM and DCM heart by IPC procedure. Unlike normal rat heart, IPC did not reverse mitochondrial dysfunction (determined by electron transport chain enzymes activity, ATP level, and membrane integrity, expression levels of genes like PGC-1ɑ, GSK3ß, complex I, II, and V) in DCM and DM rat heart. The present study demonstrated that IPC failed to protect I/R-challenged DCM rat heart, and the underlying pathology was associated with deteriorated mitochondrial function.

Beneficial effect of sodium thiosulfate extends beyond myocardial tissue in isoproterenol model of infarction: Implication for nootropic effects.

One of the common negative impacts in the management of acute myocardial infarction is cognitive decline. Using the rat model of isoproterenol (ISO)-induced myocardial infarction, we assessed the cardioprotective effect of sodium thiosulfate (STS) and its influence on cognition. STS treatment reduced the cardiac infarct size by 75%, injury markers (lactate dehydrogenase: 60%, creatine kinase-muscle/brain: 44%) release in the blood, maintain the heart rate within a normal range (365 ± 10 bpm) and minimize postinfarction hypertrophic changes in comparison with the ISO group. At the cellular level, the heart from these rats had reduced reactive oxygen species (ROS) (25%), caspase-9 (60%), and improved mitochondrial function (restored electron transport chain function and copy number) compared to ISO hearts. The brain of STS-treated rats also showed a reduction in ROS (45%), caspase-9 (37%), and improved mitochondrial function relative to the brain of the ISO group, particularly limited to the striatum region, and these rats showed improved cognitive ability. Predominantly, the STS treatment reduced the reference memory defects observed in comparison to rats challenged by ISO. Furthermore, elevated circulating mitochondrial DNA and ATP were found in ISO-challenged rats, which indicate the cardiac mitochondria linked damage-associated patterns were restored to the sham level when pretreated with STS. We found increased H2 S, a well-known metabolite of STS with a neuroprotective role in the brain after STS administration, hinting at a possible secondary defense mechanism. In conclusion, the STS mediated cardioprotection and its nootropic effects are primarily mediated via the improvement of mitochondrial function and reduction of oxidative stress.

Streptozotocin-induced type II diabetic rat administered with nonobesogenic high-fat diet is highly susceptible to myocardial ischemia-reperfusion injury: An insight into the function of mitochondria.

RATIONALE: Our recent study suggested that ischemia-reperfusion (I/R) induced oxidative stress was minimal in the rat heart during initial stage of diabetes and the one that progressed to diabetic cardiomyopathy (DCM), despite having higher infarct and low cardiac performance. Mitochondrial dysfunction is an important mediator for adverse outcome in rat heart affected with diabetes, which is also a potential contributor for the cardiac reperfusion injury. OBJECTIVE: The current study aims to evaluate the susceptibility of diabetes heart with or without myopathy to I/R injury and its influence on cardiac mitochondrial function. METHODS AND RESULTS: Male Wistar rats (3 weeks old) were fed with high-fat diet for 8 weeks followed by diabetes mellitus (DM) induction via streptozotocin (35 mg/kg body weight) and maintained for further 4 weeks. The animal displayed cardiomyopathy characteristics like hypertrophy, fibrosis, and insulin resistance-termed diabetic cardiomyopathy (DCM). To study the specific effect of DCM on I/R, we included diabetic rats without cardiomyopathy. Induction of I/R in different groups suggested higher vulnerability to injury in DCM rat hearts than DM and normal (measured via hemodynamics, triphenyltetrazolium chloride stain, and apoptotic markers). Mitochondrial function at the subpopulation level was evaluated with respect to adenosine triphosphate (ATP) concentration, membrane potential, swelling behavior, and oxidative stress, wherein the results confirmed I/R-induced mitochondrial dysfunction. Unlike normal heart, DM, and DCM heart challenged to I/R exhibited altered ATP producing capacity among subsarcolemmal and interfibrillar mitochondria. CONCLUSION: The above results suggest that mitochondrial changes associated with diabetes and cardiomyopathy significantly contribute to the adverse outcome of I/R injury.

Addressing the alterations in cerebral ischemia-reperfusion injury on the brain mitochondrial activity: A possible link to cognitive decline.

Studies suggest that different anatomical regions in a normal brain show distinct mitochondrial physiology. But the pathological role of mitochondria during ischemia-reperfusion injury (IRI) from these brain regions are yet to be addressed. The objective of this study is to identify mitochondrial perturbations from the brain regions of cortex and striatum, exposed to IRI and their correlation with cognition. A rat model of bilateral carotid artery occlusion was used to induce ischemia (15 min and 30 min) followed by reperfusion of varying time points (15 min, 30 min, 4 h and 24 h). It was evident from the results that ischemia (30 min) caused changes in cerebral histology which was aggravated upon reperfusion. Upon examining the mitochondria, ischemia significantly reduced the ETC enzyme activity (complex-I and II) and ATP level in the striatum. Reperfusion further aggravated this decline by 4 h. Following 24 h reperfusion, the functional activity of ETC recovered in the striatum but not in the cortex. The complex-I, II activity and ATP level significantly declined by 24 h reperfusion in the cortex. Cortical mitochondria showed significantly reduced antioxidant enzyme activities (catalase and GPx) by the end of 24 h reperfusion. The implication of cellular events was noted as a decline in the cortex related cognitive performance in radial arm maze and Morris water maze tests. The susceptibility of mitochondria to IRI is different across the brain regions (striatum > cortex). Hence the observed loss of mitochondrial energy metabolism might be a contributing factor for cognitive decline in IRI.

Preconditioning the rat heart with sodium thiosulfate preserved the mitochondria in response to ischemia-reperfusion injury.

Sodium thiosulfate preconditioning (SIPC) was recently reported to be cardioprotective due to its ability to inhibit caspase-3 activation, chelate calcium ions and scavenge free radicals. However, the rationale behind its ability to improve the contractility of isolated rat heart challenged with ischemia-reperfusion injury (IR) is not well understood. As mitochondrial preservation is implicated in cardioprotection against IR, the present study was conceived to identify whether the cardioprotective effects of SIPC is associated with mitochondrial preservation. Using the isolated Langendorff rat heart model, 1 mM sodium thiosulfate (STS) was used to precondition the rat heart before IR and was used to study its effect on cardiac mitochondria. The IR heart experienced a ventricular contractile dysfunction that was improved by SIPC. Upon assessing in-gel the ATP synthetic capacity of mitochondria from IR heart, there was a significant decline, while in SIPC it was well preserved close to sham. As a sustained flow of electrons through the ETC and well-integrated mitochondria are the prerequisites for ATP synthesis, SIPC improved the activities of ETC complex enzymes (I-IV), which was reflected from the preserved ultrastructure of the mitochondria as analyzed from electron-microscopy in the treated rat hearts. This observation was coherent with the elevated expression of PGC1α (20%), a critical regulator of ATP production, which increased the mitochondrial copy number as well in the STS treated heart compared to IR. In conclusion, mitochondria might be a critical target for SIPC mediated cardioprotection against IR.

Sodium Thiosulfate Preconditioning Ameliorates Ischemia/Reperfusion Injury in Rat Hearts Via Reduction of Oxidative Stress and Apoptosis.

PURPOSE: Sodium thiosulfate (STS) has of late been proven efficacious in models of urolithiasis and vascular calcification. However, its cardiovascular effects on ischemia reperfusion injury (IR) have not been revealed. Being an antioxidant and calcium chelator, it is assumed to play a vital role in IR as ROS production and calcium overload are major perpetrators of IR injury. METHODS: The cardioprotective effect of STS was evaluated in vitro using H9C2 cardiomyocytes and in vivo using both isolated rat heart and intact left anterior descending artery (LAD) occlusion models of ischemia reperfusion injury. Finally, in silico tools were utilized to establish its possible mode of action. Myocardial injury markers and expression of apoptotic proteins were studied along with myocardial histopathology. RESULTS: STS of 1 mM recovered H9C2 cells from glucose oxidase/catalase-induced apoptosis. The isolated rat heart treated with STS prior to IR injury improved its hemodynamics and reduced the infarct size to 9%. This was supported by the absence of derangement of cardiac fibers from H&E stained section of LAD-occluded rats. Plasma troponin levels decreased by 15% compared to IR and the myocardium showed diminished apoptotic proteins. An in silico docking analysis revealed higher binding affinity of STS for caspase-3 with a binding energy of - 60.523 kcal/mol for the complex. CONCLUSION: The effectiveness of STS as a cardioprotective agent is attributed to the reduction of apoptosis by binding to the active site of caspase-3 in silico, which was substantiated by the reduced expression of caspase-3 and poly ADP ribose polymerase levels.

Erythrocyte Membrane Bound ATPase and Antioxidant Enzyme Changes Associated with Vascular Calcification is Reduced by Sodium Thiosulfate.

Sodium thiosulfate (STS), a cyanide antidote has been reported to possess antioxidant and calcium chelation effects, useful for the treatment of renal failure due to vascular calcification and urolithiasis. The present study investigated the in vivo modulatory effects of STS on erythrocyte calcium, phosphorous levels, lipid peroxidation, antioxidant enzyme and membrane ATPase activities (Ca2+, Na+K+, Mg2+ and 5'' nucleotidase) in an adenine induced model of vascular calcification in rats. Adenine (0.75%) was supplemented through the diet for 28 days, which resulted in significantly (P < 0.05) increased circulating calcium and phosphorous product and oxidative stress within the RBCs, as measured from lipid peroxidation and reduced antioxidant enzymes. The membrane ATPase activities were altered (increased Ca2+, Na+K+ ATPase and decreased Mg+ ATPase, 5' nucleotidase) compared to the rats fed on normal diet. STS (400 mg/kg) given orally was effective in establishing a normalcy in the RBC alterations. This effect was more pronounced, when STS was given from day 28 to day 49 after induction of calcification, instead of day 0 to day 28. These findings may benefit to evaluate the effectiveness of STS therapy in patients with chronic renal failure associated with increased circulating calcium and phosphorous product that leads to stiffening of vascular smooth muscles of aorta, due to calcium deposition.

Studying inhibition of calcium oxalate stone formation: an in vitro approach for screening hydrogen sulfide and its metabolites.

PURPOSE: Calcium oxalate urolithiasis is one of the most common urinary tract diseases and is of high prevalence. The present study proposes to evaluate the antilithiatic property of hydrogen sulfide and its metabolites like thiosulfate & sulfate in an in vitro model. MATERIALS AND METHODS: The antilithiatic activity of sodium hydrogen sulfide (NaSH), sodium thiosulfate (Na(2)S(2)O(3)) and sodium sulfate (Na(2)SO(4)) on the kinetics of calcium oxalate crystal formation was investigated both in physiological buffer and in urine from normal and recurrent stone forming volunteers. The stones were characterized by optical and spectroscopic techniques. RESULTS: The stones were characterized to be monoclinic, prismatic and bipyramidal habit which is of calcium monohydrate and dihydrate nature. The FTIR displayed fingerprint corresponding to calcium oxalate in the control while in NaSH treated, S=O vibrations were visible in the spectrum. The order of percentage inhibition was NaSH>Na(2)S(2)O(3)>Na(2)SO(4). CONCLUSION: Our study indicates that sodium hydrogen sulfide and its metabolite thiosulfate are inhibitors of calcium oxalate stone agglomeration which makes them unstable both in physiological buffer and in urine. This effect is attributed to pH changes and complexing of calcium by S(2)O(3)(2)-and SO(4)(2)- moiety produced by the test compounds.

Effect of mitochondrial potassium channel on the renal protection mediated by sodium thiosulfate against ethylene glycol induced nephrolithiasis in rat model.

PURPOSE: Sodium thiosulfate (STS) is clinically reported to be a promising drug in preventing nephrolithiasis. However, its mechanism of action remains unclear. In the present study, we investigated the role of mitochondrial KATP channel in the renal protection mediated by STS. MATERIALS AND METHODS: Nephrolithiasis was induced in Wistar rats by administrating 0.4% ethylene glycol (EG) along with 1% ammonium chloride for one week in drinking water followed by only 0.75% EG for two weeks. Treatment groups received STS, mitochondrial KATP channel opener and closer exclusively or in combination with STS for two weeks. RESULTS: Animals treated with STS showed normal renal tissue architecture, supported by near normal serum creatinine, urea and ALP activity. Diazoxide (mitochondria KATP channel opening) treatment to the animal also showed normal renal tissue histology and improved serum chemistry. However, an opposite result was shown by glibenclamide (mitochondria KATP channel closer) treated rats. STS administered along with diazoxide negated the renal protection rendered by diazoxide alone, while it imparted protection to the glibenclamide treated rats, formulating a mitochondria modulated STS action. CONCLUSION: The present study confirmed that STS render renal protection not only through chelation and antioxidant effect but also by modulating the mitochondrial KATP channel for preventing urolithiasis.

Investigating the temporal link between PM2.5 exposure and acceleration of myocardial ischemia reperfusion injury: Emphasizing the hazardous presence of metals in inhaled air.

Exposure to PM2.5 is widely acknowledged to induce cardiotoxic effects, leading to decreased myocardial tolerance to revascularization procedures and subsequent ischemia reperfusion injury (IR). However, the temporal relationship between PM2.5 exposure and vulnerability to IR, along with the underlying mechanisms, remains unclear and is the focus of this study. Female Wistar rats were exposed to PM2.5 at a concentration of 250 µg/m³ for 3 h daily over varying durations (7, 14, and 21 days), followed by IR induction. Our results demonstrated a significant increase in cardiac injury, as evidenced by increased infarct size and elevated cardiac injury markers, starting from day 14 of PM2.5 exposure, accompanied by declined cardiac function. These adverse effects were associated with apoptosis and impaired mitochondrial function, including reduced bioenergetics, mitochondrial DNA copy number and quality control mechanisms, along with inactivation of the PI3K/AKT/AMPK signalling pathways. Furthermore, analysis of myocardial tissue revealed elevated metal accumulation, particularly within mitochondria. Chelation of PM2.5 -associated metals using EDTA significantly mitigated the toxic effects on cardiac IR pathology, as confirmed in both rat myocardium and H9c2 cells. These findings suggest that metals in PM2.5 play a crucial role in inducing cardiotoxicity, impairing myocardial resilience to stress through mitochondrial accumulation and dysfunction.

Ischemic preconditioning modulates the DNA methylation process of the rat heart to provide tolerance to withstand ischemia reperfusion injury and its associated mitochondrial dysfunction.

DNA methylation plays a crucial role in the pathogenesis of myocardial ischemia reperfusion injury(I/R) and the I/R injury can be combated effectively by ischemia preconditioning (IPC), but the role is DNA methylation in this process is unknown. In this study, we uncovered the role of ischemic preconditioning (IPC)- mediated cardioprotection of rat myocardium by using a Langendorff rat heart model with 30 min of ischemia followed by 60 min of reperfusion. Heart conditioned with short cycles of ischemia and reperfusion (IPC procedure) prior to I/R protocol significantly reduced the I/R-induced global DNA hypermethylation level by 32% and the DNMT activity by 33% while rendering cardioprotection. Blocking the PI3K pathway via wortmannin not only negates the cardio-protection by IPC, but also increases the methylation of DNA by 75%. Besides, the correlation analysis showed a negative relationship between PI3K gene expression and the global DNA methylation level (r = - 0.8690, p = 0.0419) in IPC-treated rat hearts. Moreover, the global level DNA hypomethylation induced by IPC exhibited a regulatory effect on the genes involved in I/R pathology mediators like apoptosis (Caspase3), mitochondrial function (PGC 1α, TFAM, ND1) and oxidative stress (CuZnSOD, SOD2), and their corresponding function. The present study results provide novel evidence for the involvement of DNA methylation in the IPC procedure, and suggest DNA methylation as one of the potential therapeutic targets regulated by ischemic preconditioning in rat hearts subjected to ischemia reperfusion. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-024-03965-0.