Follistatin protein enhances satellite cell counts in reinnervated muscle

Background Muscle recovery following peripheral nerve repair is sup-optimal. Follistatin (FST), a potent muscle stimulant, enhances muscle size and satellite cell counts following reinnervation when administered as recombinant FST DNA via viral vectors. Local administration of recombinant FST protein, if effective, would be more clinically translatable but has yet to be investigated following muscle reinnervation. Objective  The aim of this study is to assess the effect of direct delivery of recombinant FST protein on muscle recovery following muscle reinnervation. Materials and Methods  In total, 72 Sprague-Dawley rats underwent temporary (3 or 6 months) denervation or sham denervation. After reinnervation, rats received FST protein (isoform FS-288) or sham treatment via a subcutaneous osmotic pump delivery system. Outcome measures included muscle force, muscle histomorphology, and FST protein quantification. Results  Follistatin treatment resulted in smaller muscles after 3 months denervation ( p  = 0.019) and reduced force after 3 months sham denervation ( p  < 0.001). Conversely, after 6 months of denervation, FST treatment trended toward increased force output ( p  = 0.066). Follistatin increased satellite cell counts after denervation ( p  < 0.001) but reduced satellite cell counts after sham denervation ( p  = 0.037). Conclusion  Follistatin had mixed effects on muscle weight and force. Direct FST protein delivery enhanced satellite cell counts following reinnervation. The positive effect on the satellite cell population is intriguing and warrants further investigation

Role of Antipsychotics-Induced Mitochondrial Dysfunction in Increased Cardiovascular Risk

Tesis Doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Medicina, Departamento de Bioquímica. Fecha de Lectura: 23-01-2023Esta Tesis tiene embargado el acceso al texto completo hasta el 23-07-2024Second generation antipsychotics (SGAs) are preferred during chronic therapy for schizophrenic and psychotic patients due to less extrapyramidal side effects. On the other hand, they are associated with increased risk of cardiovascular diseases (CVDs), which is major cause of mortality among these patients. The SGAs are associated with metabolic side effects, where Olanzapine (Ola) is associated with increased metabolic risks; whereas Ari (Ari), less vigorously studied so far, is regarded as relatively safer. However, clear cause-effect relationships explaining such diverse metabolic profiles are yet to be established. Recently many drugs, including some SGAs, are found to induce mitochondrial dysfunction, which might be linked to metabolic and cardiovascular risks. Hence, we decided to evaluate role of mitochondrial function in development of cardiovascular risk due to SGAs in order to determine whether it can be used as risk predictor. In light of this, we investigated mitochondrial distribution of Ola and Ari in mice, as well as their effects on mitochondrial respiration clinically. We studied effects of subacute and chronic treatment with Ari and Ola on metabolic and cardiovascular functions in wild type (WT) and PGC-1α-deficient mice (KO), a model of mitochondrial dysfunction. We observed that both Ari and Ola enter mitochondria at significant levels. Ari reduced mitochondrial respiration efficiency, mitochondrial content, and cristae density severely and irreversibly; whereas Ola affected mitochondrial respiration and cristae density with compensatory recovery in mitochondrial content and respiration. Ari reduced whole body respiration and energy expenditure in vivo which was persistent and evident in both WT and KO mice, whereas Ola reduced respiration and energy expenditure which was dependent on genotype and tends to recover with time. In line with these observations, Ari found to cause glucose intolerance earlier than Ola with the effect being earlier and severe in KO mice compared to WT mice. Such metabolic changes coincided with profibrotic changes in cardiovascular structure especially in KO mice treated with Ari and Ola for long duration, which ultimately affects macrovascular function, and cardiac recovery after ischemia-reperfusion injury. Such detrimental effects of Ari and Ola on cardiovascular structure and functions were found to be dependent on mitochondrial function. The results indicate protective role of PGC-1α against drug-induced toxicity in cardiovascular system. These results could encourage potential use of drug screening for their mitochondrial effects during drug development. Also, evaluation of mitochondrial function in patients before and during therapy could be used to predict the risk of developing cardiovascular diseases and to guide selection of therapy as a part of personalised medicineThis work was supported by the grant ‘Horizon 2020 Marie Sklodowska-Curie Innovative Training Network of EU (Grant# 721236)’ to Dr. María Monsalve Pérez and Gaurangkumar Pate

Experimental Data of the Doctoral Thesis of Gaurang Patel (ESR4. ITN TREATMENT)

DATA EXPLANATION: [Folder 1]. Analysis of Drug concentration in plasma and mitochondria (1h and 24 h). The xls file shows the statistics analysis of the detected concentration in plasma and mitochondrial samples after 1 and 24 hours after adding the treatment by aripripazole (Ari), dehydroaripripazole (Dehydroari) or olanzapine (Ola). The .pzfx file can be opened with GraphPad Prism 8 XML Project and shows the decreased in the concentrations across the time in plasma samples. [Folder 2]. Seahorse Data Healthy volunteer study. The two xls files show (1) the raw data obtained from the measurement of oxygen flux, oxygen consumption rate [OCR] and proton flux, extracellular acidification rate [ECAR], in the medium immediately surrounding the cells in a microplate in a Seahorse analyzer and (2) the linear regression between these data. And, the .pzfx files can be opened with GraphPad Prism 8 XML Project and show the relationships between some of the parameters showed in xls files.Funding agency and Grant Agreetment Number : European Comission ITN Treatment H2020-MSCA-ITN-721236Peer reviewe

Role of mitochondrial dysfunction in NAFL. Macrovascular alterations in PGC-1alfa knock-out mice

Trabajo presentado a la reunión de la Red CONSOLREDOX, que lleva por titulo "Redox Biology as a major drive to the understanding of pathophysiology", celebrada en Salamanca (España) del 23 al 25 de abril de 2018.Peer Reviewe

SGAs induce mitochondrial dysfunction in vascular endothelial cells

Resumen del póster presentado al 42nd Congress of the Spanish Society of Biochemistry and Molecular Biology (SEBBM), celebrado en Madrid del 16 al 19 de julio de 2019.Chronic administration of pharmaceutical drugs results in the increased risk of cardiovascular disease (CVD). However, the mechanisms involved have not been elucidated. Recent studies show that many drugs inhibit mitochondrial electron transport chain. Second Generation Antipsychotics (SGAs) are known because their chronic administration results in an increased incidence of CVD. The present study aimed to elucidate the impact of the SGAs Aripiprazole (Ari) and Olanzapine (Ola) on vascular endothelial cell bioenergetics. As a model system, we used bovine aortic endothelial cells (BAEC) cultured in non-oxidative (25mM glucose) or oxidative (5mM galactose) conditions and treated them with 300nM and 3uM Ari, Ola. The evaluation of oxygen consumption rates (OCR) showed that basal respiration and maximal respiratory capacity were drastically reduced in a 3h exposure to 3uM concentration of Ari and Ola. At 24h Ola treated cells in high glucose media had recovered to control levels, while respiration reached higher levels than in control cells in galactose media, suggesting that Ola had an hormetic effect in this condition. However, the respiration of Ari treated cells remained below control levels after 24h which suggested a lower capacity to induce compensatory mechanism. Analysis of the expression levels of PGC-1a and cyt c, showed that Ari and Ola induced an early upregulation of PGC-1a levels and activity (6h), that resulted in an induction of cyt c that peaked at 24h. After 24h, the capacity to induce PGC-1a levels was lost in Ari, but not in Ola treated cells, again suggesting a higher recovery capacity for Ola treated cells. We tested the effect on cellular redox balance analyzing the mRNA and protein levels of several antioxidants. In the treatment with Ola was an early induction (6h) of genes that regulate gluthation synthesis (gss, gclc), while Ari induced a downregulation of those genes at 24h. Furthermore, the levels of proteins like catalase and thioredoxin reductase 2 were drastically downregulated after 6h of Ari treatment, but not Ola, suggesting a more severe loss of antioxidant capacity in Ari treated cells. In sum, Ola and Ari treatment result in an inhibition of mitochondrial OCR, but Ari induced a more enduring alteration of cellular bioenergetics that may result in the induction of endothelial dysfunction.Peer reviewe

Role of mitochondria in atypical antipsychotics induced cardiovascular disorders

Resumen del trabajo presentado al 42nd Congress of the Spanish Society of Biochemistry and Molecular Biology (SEBBM), celebrado en Madrid del 16 al 19 de julio de 2019.Schizophrenic patients require life-long treatment with atypical antipsychotics, also called as second generation antipsychotics (SGAs), like Olanzapine (Ola) being the most common. Long-term use of SGAs administration is associated with secondary metabolic effects like weight gain, dyslipidemia, and hyperglycemia which are considered as major risk factors for cardiovascular diseases (CVDs). However, the mechanisms involved have not been elucidated. Taking into account the central role played by mitochondrial dysfunction in the development of CVDs, we aimed to elucidate the role played by the possible interference of Olanzapine (Ola) and Aripiprazole (Ari) with mitochondrial function in CVDs. To that end, we used PGC-1a deficient (KO) mice as a model system of systemic mitochondrial dysfunction. We have compared the effects of subacute (ip, 28 days) and chronic (diet, 6 months) administration of Ari and Ola (5 mg/kg) on vascular reactivity of aortic rings from PGC-1a WT and KO mice tested ex-vivo. The effects of the treatment on the systemic use of oxygen was determined at 1 month, 3 months and 6 months of treatment using indirect calorimetry. We have found that Ari has more potent effect of than Ola, while both inhibited OCR in a PGC-1a dependent manner, supporting the hypothesis of induced mitochondrial dysfunction in treated mice. Vascular reactivity analysis showed that Ari enhanced the vascular constriction response to angiotensin II (Ang II) and endothelin 1 (ET1) and altered the dose response curve during relaxation to acetylcholine (Ach), whereas while Ola increased the response to norepinephrine (NE) at 1 and 6 months in WT mice. These effects were associated with corresponding changes in the gene expression pattern of the Ang II receptor AT1-R and the ET1 receptor ETA-R. These changes indicated a higher vascular toxicity of Ari that was dependent on mitochondrial function and affected in particular the response to Ang II and ET1 leading to hyper-reactivity. In sum, both Ari and Ola administration in mice reduce oxidative metabolism with Ari treatment being more potent. The observation that these changes do not occur in PGC-1a KO mice support the notion that they are dependent on the inhibition of oxidative metabolism.Peer reviewe

Melatonin Effects on Non-Alcoholic Fatty Liver Disease Are Related to MicroRNA-34a-5p/Sirt1 Axis and Autophagy

Melatonin, an indole produced by pineal and extrapineal tissues, but also taken with a vegetarian diet, has strong anti-oxidant, anti-inflammatory and anti-obesogenic potentials. Non-alcoholic fatty liver disease (NAFLD) is the hepatic side of the metabolic syndrome. NAFLD is a still reversible phase but may evolve into steatohepatitis (NASH), cirrhosis and carcinoma. Currently, an effective therapy for blocking NAFLD staging is lacking. Silent information regulator 1 (SIRT1), a NAD+ dependent histone deacetylase, modulates the energetic metabolism in the liver. Micro-RNA-34a-5p, a direct inhibitor of SIRT1, is an emerging indicator of NAFLD grading. Thus, here we analyzed the effects of oral melatonin against NAFLD and underlying molecular mechanisms, focusing on steatosis, ER stress, mitochondrial shape and autophagy. Male C57BL/6J (WT) and SIRT1 heterozygous (HET) mice were placed either on a high-fat diet (58.4% energy from lard) (HFD) or on a standard maintenance diet (8.4% energy from lipids) for 16 weeks, drinking melatonin (10 mg/kg) or not. Indirect calorimetry, glucose tolerance, steatosis, inflammation, ER stress, mitochondrial changes, autophagy and microRNA-34a-5p expression were estimated. Melatonin improved hepatic metabolism and steatosis, influenced ER stress and mitochondrial shape, and promoted autophagy in WT HFD mice. Conversely, melatonin was ineffective in HET HFD mice, maintaining NASH changes. Indeed, autophagy was inconsistent in HET HFD or starved mice, as indicated by LC3II/LC3I ratio, p62/SQSTM1 and autophagosomes estimation. The beneficial role of melatonin in dietary induced NAFLD/NASH in mice was related to reduced expression of microRNA-34a-5p and sterol regulatory element-binding protein (SREBP1) but only in the presence of full SIRT1 availability