Pub Date : 2024-04-17DOI: 10.1016/j.biochi.2024.04.007
Martina Gyimesi , Lotta E. Oikari , Chieh Yu , Heidi G. Sutherland , Dale R. Nyholt , Lyn R. Griffiths , Andre J. Van Wijnen , Rachel K. Okolicsanyi , Larisa M. Haupt
Stem cell therapies hold promise in addressing the burden of neurodegenerative diseases with human embryonic neural stem cells (hNSC-H9s) and bone marrow-derived human mesenchymal stem cells (hMSCs) as viable candidates. The induction of hMSC neurospheres (hMSC-IN) generate a more lineage-restricted common neural progenitor-like cell population, potentially tunable by heparan sulfate proteoglycans (HSPGs). We examined CpG (5 mC) site methylation patterns using Illumina Infinium 850 K EPIC arrays in hNSC-H9, hMSCs and hMSC-IN cultures with HSPG agonist heparin at early and late phases of growth. We identified key regulatory CpG sites in syndecans (SDC2; SDC4) that potentially regulate gene expression in monolayers. Unique hMSC-IN hypomethylation in glypicans (GPC3; GPC4) underscore their significance in neural lineages with Sulfatase 1 and 2 (SULF1 & 2) CpG methylation changes potentially driving the neurogenic shift. hMSC-INs methylation levels at SULF1 CpG sites and SULF2:cg25401628 were more closely aligned with hNSC-H9 cells than with hMSCs. We further suggest SOX2 regulation governed by lncSOX2-Overall Transcript (lncSOX2-OT) methylation changes with preferential activation of ENO2 over other neuronal markers within hMSC-INs. Our findings illuminate epigenetic dynamics governing neural lineage commitment of hMSC-INs offering insights for targeted mechanisms for regenerative medicine and therapeutic strategies.
{"title":"CpG methylation changes in human mesenchymal and neural stem cells in response to in vitro niche modifications","authors":"Martina Gyimesi , Lotta E. Oikari , Chieh Yu , Heidi G. Sutherland , Dale R. Nyholt , Lyn R. Griffiths , Andre J. Van Wijnen , Rachel K. Okolicsanyi , Larisa M. Haupt","doi":"10.1016/j.biochi.2024.04.007","DOIUrl":"10.1016/j.biochi.2024.04.007","url":null,"abstract":"<div><p>Stem cell therapies hold promise in addressing the burden of neurodegenerative diseases with human embryonic neural stem cells (hNSC-H9s) and bone marrow-derived human mesenchymal stem cells (hMSCs) as viable candidates. The induction of hMSC neurospheres (hMSC-IN) generate a more lineage-restricted common neural progenitor-like cell population, potentially tunable by heparan sulfate proteoglycans (HSPGs). We examined CpG (5 mC) site methylation patterns using Illumina Infinium 850 K EPIC arrays in hNSC-H9, hMSCs and hMSC-IN cultures with HSPG agonist heparin at early and late phases of growth. We identified key regulatory CpG sites in syndecans (<em>SDC2; SDC4</em>) that potentially regulate gene expression in monolayers. Unique hMSC-IN hypomethylation in glypicans (<em>GPC3</em>; <em>GPC4)</em> underscore their significance in neural lineages with Sulfatase 1 and 2 (<em>SULF1</em> & <em>2</em>) CpG methylation changes potentially driving the neurogenic shift. hMSC-INs methylation levels at <em>SULF1</em> CpG sites and <em>SULF2</em>:cg25401628 were more closely aligned with hNSC-H9 cells than with hMSCs. We further suggest <em>SOX2</em> regulation governed by lncSOX2-Overall Transcript <em>(lncSOX2-OT)</em> methylation changes with preferential activation of <em>ENO2</em> over other neuronal markers within hMSC-INs. Our findings illuminate epigenetic dynamics governing neural lineage commitment of hMSC-INs offering insights for targeted mechanisms for regenerative medicine and therapeutic strategies.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2024-04-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0300908424000828/pdfft?md5=1d288c983293875ff2a9342fcffc1843&pid=1-s2.0-S0300908424000828-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140793340","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-04-10DOI: 10.1016/j.biochi.2024.04.002
Gimena Salas, Alen A. Litta, Anabela C. Medeot, Virginia S. Schuck, Romina B. Andermatten, Gisel S. Miszczuk, Nadia Ciriaci, Ma Valeria Razori, Ismael R. Barosso, Enrique J. Sánchez Pozzi, Marcelo G. Roma, Cecilia L. Basiglio, Fernando A. Crocenzi
The endogenous metabolite of estradiol, estradiol 17β-D-glucuronide (E17G), is considered the main responsible of the intrahepatic cholestasis of pregnancy. E17G alters the activity of canalicular transporters through a signaling pathway-dependent cellular internalization, phenomenon that was attributed to oxidative stress in different cholestatic conditions. However, there are no reports involving oxidative stress in E17G-induced cholestasis, representing this the aim of our work. Using polarized hepatocyte cultures, we showed that antioxidant compounds prevented E17G-induced Mrp2 activity alteration, being this alteration equally prevented by the NADPH oxidase (NOX) inhibitor apocynin. The model antioxidant N-acetyl-cysteine prevented, in isolated and perfused rat livers, E17G-induced impairment of bile flow and Mrp2 activity, thus confirming the participation of reactive oxygen species (ROS) in this cholestasis. In primary cultured hepatocytes, pretreatment with specific inhibitors of ERK1/2 and p38MAPK impeded E17G-induced ROS production; contrarily, NOX inhibition did not affect ERK1/2 and p38MAPK phosphorylation. Both, knockdown of p47phox by siRNA and preincubation with apocynin in sandwich-cultured rat hepatocytes significantly prevented E17G-induced internalization of Mrp2, suggesting a crucial role for NOX in this phenomenon. Concluding, E17G-induced cholestasis is partially mediated by NOX-generated ROS through internalization of canalicular transporters like Mrp2, being ERK1/2 and p38MAPK necessary for NOX activation.
{"title":"NADPH oxidase-generated reactive oxygen species are involved in estradiol 17ß-d-glucuronide-induced cholestasis","authors":"Gimena Salas, Alen A. Litta, Anabela C. Medeot, Virginia S. Schuck, Romina B. Andermatten, Gisel S. Miszczuk, Nadia Ciriaci, Ma Valeria Razori, Ismael R. Barosso, Enrique J. Sánchez Pozzi, Marcelo G. Roma, Cecilia L. Basiglio, Fernando A. Crocenzi","doi":"10.1016/j.biochi.2024.04.002","DOIUrl":"https://doi.org/10.1016/j.biochi.2024.04.002","url":null,"abstract":"<div><p>The endogenous metabolite of estradiol, estradiol 17β-D-glucuronide (E17G), is considered the main responsible of the intrahepatic cholestasis of pregnancy. E17G alters the activity of canalicular transporters through a signaling pathway-dependent cellular internalization, phenomenon that was attributed to oxidative stress in different cholestatic conditions. However, there are no reports involving oxidative stress in E17G-induced cholestasis, representing this the aim of our work. Using polarized hepatocyte cultures, we showed that antioxidant compounds prevented E17G-induced Mrp2 activity alteration, being this alteration equally prevented by the NADPH oxidase (NOX) inhibitor apocynin. The model antioxidant <em>N</em>-acetyl-cysteine prevented, in isolated and perfused rat livers, E17G-induced impairment of bile flow and Mrp2 activity, thus confirming the participation of reactive oxygen species (ROS) in this cholestasis. In primary cultured hepatocytes, pretreatment with specific inhibitors of ERK1/2 and p38MAPK impeded E17G-induced ROS production; contrarily, NOX inhibition did not affect ERK1/2 and p38MAPK phosphorylation. Both, knockdown of p47phox by siRNA and preincubation with apocynin in sandwich-cultured rat hepatocytes significantly prevented E17G-induced internalization of Mrp2, suggesting a crucial role for NOX in this phenomenon. Concluding, E17G-induced cholestasis is partially mediated by NOX-generated ROS through internalization of canalicular transporters like Mrp2, being ERK1/2 and p38MAPK necessary for NOX activation.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140555269","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Alzheimer's disease (AD) and related dementias constitute an important global health challenge. Detailed understanding of the multiple molecular mechanisms underlying their pathogenesis constitutes a clue for the management of the disease. Kallikrein-related peptidases (KLKs), a lead family of serine proteases, have emerged as potential biomarkers and therapeutic targets in the context of AD and associated cognitive decline. Hence, KLKs were proposed to display multifaceted impacts influencing various aspects of neurodegeneration, including amyloid-beta aggregation, tau pathology, neuroinflammation, and synaptic dysfunction. We propose here a comprehensive survey to summarize recent findings, providing an overview of the main kallikreins implicated in AD pathophysiology namely KLK8, KLK6 and KLK7. We explore the interplay between KLKs and key AD molecular pathways, shedding light on their significance as potential biomarkers for early disease detection. We also discuss their pertinence as therapeutic targets for disease-modifying interventions to develop innovative therapeutic strategies aimed at halting or ameliorating the progression of AD and associated dementias.
阿尔茨海默病(AD)和相关痴呆症是全球健康面临的一项重要挑战。详细了解这些疾病发病的多种分子机制是治疗这些疾病的线索。Kallikrein相关肽酶(KLKs)是丝氨酸蛋白酶的一个主要家族,已成为AD和相关认知能力下降的潜在生物标志物和治疗靶点。因此,KLKs 被认为会对神经退行性病变的各个方面产生多方面的影响,包括淀粉样蛋白-β聚集、tau 病理学、神经炎症和突触功能障碍。我们在此提出一项全面调查,总结最近的研究结果,概述与 AD 病理生理学有关的主要 Kallikreins,即 KLK8、KLK6 和 KLK7。我们探讨了 KLKs 与关键的 AD 分子通路之间的相互作用,揭示了它们作为早期疾病检测的潜在生物标记物的意义。我们还讨论了它们作为疾病改变干预治疗靶点的相关性,以开发创新的治疗策略,阻止或改善注意力缺失症和相关痴呆症的进展。
{"title":"Kallikrein-related peptidase's significance in Alzheimer's disease pathogenesis: A comprehensive survey","authors":"Rilès Boumali, Laureline Urli, Meriem Naim, Feryel Soualmia, Kiyoka Kinugawa, Isabelle Petropoulos, Chahrazade El Amri","doi":"10.1016/j.biochi.2024.04.001","DOIUrl":"https://doi.org/10.1016/j.biochi.2024.04.001","url":null,"abstract":"Alzheimer's disease (AD) and related dementias constitute an important global health challenge. Detailed understanding of the multiple molecular mechanisms underlying their pathogenesis constitutes a clue for the management of the disease. Kallikrein-related peptidases (KLKs), a lead family of serine proteases, have emerged as potential biomarkers and therapeutic targets in the context of AD and associated cognitive decline. Hence, KLKs were proposed to display multifaceted impacts influencing various aspects of neurodegeneration, including amyloid-beta aggregation, tau pathology, neuroinflammation, and synaptic dysfunction. We propose here a comprehensive survey to summarize recent findings, providing an overview of the main kallikreins implicated in AD pathophysiology namely KLK8, KLK6 and KLK7. We explore the interplay between KLKs and key AD molecular pathways, shedding light on their significance as potential biomarkers for early disease detection. We also discuss their pertinence as therapeutic targets for disease-modifying interventions to develop innovative therapeutic strategies aimed at halting or ameliorating the progression of AD and associated dementias.","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140615277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-04-04DOI: 10.1016/j.biochi.2024.03.017
Hasana Baber , Arega Aghajani , B. Harold Gallimore , Cassandra Bethel , James G. Hyatt , Elizabeth F.B. King , Helen P. Price , Marissa L. Maciej-Hulme , Suat Sari , Anja Winter
Leishmaniasis is a spectrum of conditions caused by infection with the protozoan Leishmania spp. parasites. Leishmaniasis is endemic in 98 countries around the world, and resistance to current anti-leishmanial drugs is rising. Our work has identified and characterised a previously unstudied galactokinase-like protein (GalK) in Leishmania donovani, which catalyses the MgATP-dependent phosphorylation of the C-1 hydroxyl group of d-galactose to galactose-1-phosphate. Here, we report the production of the catalytically active recombinant protein in E. coli, determination of its substrate specificity and kinetic constants, as well as analysis of its molecular envelope using in solution X-ray scattering. Our results reveal kinetic parameters in range with other galactokinases with an average apparent Km value of 76 μM for galactose, Vmax and apparent Kcat values with 4.46376 × 10−9 M/s and 0.021 s−1, respectively. Substantial substrate promiscuity was observed, with galactose being the preferred substrate, followed by mannose, fructose and GalNAc. LdGalK has a highly flexible protein structure suggestive of multiple conformational states in solution, which may be the key to its substrate promiscuity. Our data presents novel insights into the galactose salvaging pathway in Leishmania and positions this protein as a potential target for the development of pharmaceuticals seeking to interfere with parasite substrate metabolism.
利什曼病是由原生动物利什曼属寄生虫感染引起的一系列疾病。利什曼病在全球 98 个国家流行,目前抗利什曼病药物的抗药性正在上升。我们的工作发现并描述了唐氏利什曼原虫中一种以前未研究过的半乳糖激酶样蛋白(GalK),它能催化 MgATP 依赖性磷酸化 d-半乳糖的 C-1 羟基为半乳糖-1-磷酸。在此,我们报告了在大肠杆菌中生产具有催化活性的重组蛋白、测定其底物特异性和动力学常数以及利用溶液中 X 射线散射分析其分子包膜的情况。我们的研究结果表明,该蛋白的动力学参数与其他半乳糖激酶相同,对半乳糖的平均表观 Km 值为 76 μM,Vmax 和表观 Kcat 值分别为 4.46376 × 10-9 M/s 和 0.021 s-1。观察到底物具有很大的杂合性,半乳糖是首选底物,其次是甘露糖、果糖和 GalNAc。LdGalK 具有高度灵活的蛋白质结构,表明其在溶液中具有多种构象状态,这可能是其底物杂交性的关键所在。我们的数据提供了有关利什曼原虫半乳糖挽救途径的新见解,并将该蛋白定位为开发药物干扰寄生虫底物代谢的潜在靶点。
{"title":"Galactokinase-like protein from Leishmania donovani: Biochemical and structural characterization of a recombinant protein","authors":"Hasana Baber , Arega Aghajani , B. Harold Gallimore , Cassandra Bethel , James G. Hyatt , Elizabeth F.B. King , Helen P. Price , Marissa L. Maciej-Hulme , Suat Sari , Anja Winter","doi":"10.1016/j.biochi.2024.03.017","DOIUrl":"https://doi.org/10.1016/j.biochi.2024.03.017","url":null,"abstract":"<div><p>Leishmaniasis is a spectrum of conditions caused by infection with the protozoan <em>Leishmania</em> spp. parasites. Leishmaniasis is endemic in 98 countries around the world, and resistance to current anti-leishmanial drugs is rising. Our work has identified and characterised a previously unstudied galactokinase-like protein (GalK) in <em>Leishmania donovani</em>, which catalyses the MgATP-dependent phosphorylation of the C-1 hydroxyl group of <span>d</span>-galactose to galactose-1-phosphate. Here, we report the production of the catalytically active recombinant protein in <em>E. coli</em>, determination of its substrate specificity and kinetic constants, as well as analysis of its molecular envelope using <em>in solution</em> X-ray scattering. Our results reveal kinetic parameters in range with other galactokinases with an average apparent Km value of 76 μM for galactose, V<sub>max</sub> and apparent K<sub>cat</sub> values with 4.46376 × 10<sup>−9</sup> M/s and 0.021 s<sup>−1</sup>, respectively. Substantial substrate promiscuity was observed, with galactose being the preferred substrate, followed by mannose, fructose and GalNAc. <em>Ld</em>GalK has a highly flexible protein structure suggestive of multiple conformational states in solution, which may be the key to its substrate promiscuity. Our data presents novel insights into the galactose salvaging pathway in <em>Leishmania</em> and positions this protein as a potential target for the development of pharmaceuticals seeking to interfere with parasite substrate metabolism.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-04-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140536810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-30DOI: 10.1016/j.biochi.2024.03.012
J. Michael Conlon , Ananyaa Sridhar , Dawood Khan , Taylor S. Cunning , Jack J. Delaney , Megan G. Taggart , Nigel G. Ternan , Jérôme Leprince , Laurent Coquet , Thierry Jouenne , Samir Attoub , Milena Mechkarska
Five host-defense peptides (figainin 2PL, hylin PL, raniseptin PL, plasticin PL, and peptide YL) were isolated from norepinephrine-stimulated skin secretions of the banana tree dwelling frog Boana platanera (Hylidae; Hylinae) collected in Trinidad.
Raniseptin PL (GVFDTVKKIGKAVGKFALGVAKNYLNS.NH2) and figainin 2PL (FLGTVLKLGKAIAKTVVPMLTNAMQPKQ. NH2) showed potent and rapid bactericidal activity against a range of clinically relevant Gram-positive and Gram-negative ESKAPE + pathogens and Clostridioides difficile. The peptides also showed potent cytotoxic activity (LC50 values < 30 μM) against A549, MDA-MB-231 and HT29 human tumor-derived cell lines but appreciably lower hemolytic activity against mouse erythrocytes (LC50 = 262 ± 14 μM for raniseptin PL and 157 ± 16 μM for figainin 2PL). Hylin PL (FLGLIPALAGAIGNLIK.NH2) showed relatively weak activity against microorganisms but was more hemolytic. The glycine-leucine-rich peptide with structural similarity to the plasticins (GLLSTVGGLVGGLLNNLGL.NH2) and the non-cytotoxic peptide YL (YVPGVIESLL.NH2) lacked antimicrobial and cytotoxic activities. Hylin PL, raniseptinPL and peptide YL stimulated the rate of release of insulin from BRIN-BD11 clonal β-cells at concentrations ≥100 nM. Peptide YL was the most effective (2.3-fold increase compared with basal rate at 1 μM concentration) and may represent a template for the design of a new class of incretin-based anti-diabetic drugs.
{"title":"Multifunctional host-defense peptides isolated from skin secretions of the banana tree dwelling frog Boana platanera (Hylidae; Hylinae)","authors":"J. Michael Conlon , Ananyaa Sridhar , Dawood Khan , Taylor S. Cunning , Jack J. Delaney , Megan G. Taggart , Nigel G. Ternan , Jérôme Leprince , Laurent Coquet , Thierry Jouenne , Samir Attoub , Milena Mechkarska","doi":"10.1016/j.biochi.2024.03.012","DOIUrl":"10.1016/j.biochi.2024.03.012","url":null,"abstract":"<div><p>Five host-defense peptides (figainin 2PL, hylin PL, raniseptin PL, plasticin PL, and peptide YL) were isolated from norepinephrine-stimulated skin secretions of the banana tree dwelling frog <em>Boana platanera</em> (Hylidae; Hylinae) collected in Trinidad.</p><p>Raniseptin PL (GVFDTVKKIGKAVGKFALGVAKNYLNS.NH<sub>2</sub>) and figainin 2PL (FLGTVLKLGKAIAKTVVPMLTNAMQPKQ. NH<sub>2</sub>) showed potent and rapid bactericidal activity against a range of clinically relevant Gram-positive and Gram-negative ESKAPE <sup>+</sup> pathogens and <em>Clostridioides difficile.</em> The peptides also showed potent cytotoxic activity (LC<sub>50</sub> values < 30 μM) against A549, MDA-MB-231 and HT29 human tumor-derived cell lines but appreciably lower hemolytic activity against mouse erythrocytes (LC<sub>50</sub> = 262 ± 14 μM for raniseptin PL and 157 ± 16 μM for figainin 2PL). Hylin PL (FLGLIPALAGAIGNLIK.NH<sub>2</sub>) showed relatively weak activity against microorganisms but was more hemolytic. The glycine-leucine-rich peptide with structural similarity to the plasticins (GLLSTVGGLVGGLLNNLGL.NH<sub>2</sub>) and the non-cytotoxic peptide YL (YVPGVIESLL.NH<sub>2</sub>) lacked antimicrobial and cytotoxic activities. Hylin PL, raniseptinPL and peptide YL stimulated the rate of release of insulin from BRIN-BD11 clonal β-cells at concentrations ≥100 nM. Peptide YL was the most effective (2.3-fold increase compared with basal rate at 1 μM concentration) and may represent a template for the design of a new class of incretin-based anti-diabetic drugs.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0300908424000701/pdfft?md5=0e62b58f0bacba03f8bb7c5ea584abbd&pid=1-s2.0-S0300908424000701-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140338045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-26DOI: 10.1016/j.biochi.2024.03.014
Maria M. Oliveira , Sofia Correia , Cecilia Peirone , Marques Magalhães , Paula Oliveira , Francisco Peixoto
Ozone therapy's efficacy might stem from the regulated and mild oxidative stress resulting from ozone's interactions with various biological elements. The present work aimed to characterize the hepatic mitochondrial response to ozone treatment and its relationship with the antioxidant system response. Two groups of mice were used: one control group and another injected intraperitoneally with an O3/O2 mixture (80 ml/kg) for 5 days. Mitochondrial respiration supported by different substrates was significantly inhibited, as well as complexes I and II/III, but not complex IV. The analysis of the electron transport chain complex activity showed significant inhibitions in complexes I and II/III but not in complex IV. These inhibitions can prevent mitochondrial reactive oxygen species (ROS) production. Additionally, there was a decline in glutathione content, unaccompanied by a rise in its oxidized form. The ozone-treated groups showed a significant increase in the activity of superoxide dismutase and glutathione peroxidase, while catalase and glutathione reductase experienced no significant alterations. Adenine nucleotides increased in the ozone group, but only the increase in adenosine diphosphate is significant, so the cell's energy charge is unaffected. This study shows that mitochondria may play a crucial role in ozone treatment. However, it also highlights the need for further studies to understand the molecular mechanism.
臭氧疗法的疗效可能源于臭氧与各种生物元素相互作用所产生的调节性轻微氧化应激。本研究旨在分析肝线粒体对臭氧治疗的反应及其与抗氧化系统反应的关系。研究使用了两组小鼠:一组为对照组,另一组为腹腔注射臭氧/二氧化硫混合物(80 毫升/千克)5 天组。不同底物支持的线粒体呼吸受到明显抑制,复合体 I 和 II/III 也受到抑制,但复合体 IV 不受抑制。对电子传递链复合物活性的分析表明,复合物 I 和 II/III 受到了明显的抑制,但复合物 IV 没有受到抑制。这些抑制作用可以阻止线粒体活性氧(ROS)的产生。此外,谷胱甘肽的含量也有所下降,但其氧化形式并未随之上升。臭氧处理组的超氧化物歧化酶和谷胱甘肽过氧化物酶的活性显著增加,而过氧化氢酶和谷胱甘肽还原酶没有发生显著变化。臭氧组中腺嘌呤核苷酸增加,但只有二磷酸腺苷增加显著,因此细胞的能量电荷未受影响。这项研究表明,线粒体可能在臭氧治疗中起着至关重要的作用。不过,它也强调了进一步研究以了解分子机制的必要性。
{"title":"Impact of ozone therapy on mouse liver mitochondrial function and antioxidant system","authors":"Maria M. Oliveira , Sofia Correia , Cecilia Peirone , Marques Magalhães , Paula Oliveira , Francisco Peixoto","doi":"10.1016/j.biochi.2024.03.014","DOIUrl":"10.1016/j.biochi.2024.03.014","url":null,"abstract":"<div><p>Ozone therapy's efficacy might stem from the regulated and mild oxidative stress resulting from ozone's interactions with various biological elements. The present work aimed to characterize the hepatic mitochondrial response to ozone treatment and its relationship with the antioxidant system response. Two groups of mice were used: one control group and another injected intraperitoneally with an O<sub>3</sub>/O<sub>2</sub> mixture (80 ml/kg) for 5 days. Mitochondrial respiration supported by different substrates was significantly inhibited, as well as complexes I and II/III, but not complex IV. The analysis of the electron transport chain complex activity showed significant inhibitions in complexes I and II/III but not in complex IV. These inhibitions can prevent mitochondrial reactive oxygen species (ROS) production. Additionally, there was a decline in glutathione content, unaccompanied by a rise in its oxidized form. The ozone-treated groups showed a significant increase in the activity of superoxide dismutase and glutathione peroxidase, while catalase and glutathione reductase experienced no significant alterations. Adenine nucleotides increased in the ozone group, but only the increase in adenosine diphosphate is significant, so the cell's energy charge is unaffected. This study shows that mitochondria may play a crucial role in ozone treatment. However, it also highlights the need for further studies to understand the molecular mechanism.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0300908424000725/pdfft?md5=ec8fe957fe9a87be5067c6dab43ed873&pid=1-s2.0-S0300908424000725-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140320056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-26DOI: 10.1016/j.biochi.2024.03.016
Samuel Barbosa , Mafalda Barbosa Pedrosa , Rita Ferreira , Daniel Moreira-Gonçalves , Lúcio Lara Santos
The depletion of visceral and subcutaneous adipose tissue (AT) during chemotherapy significantly correlates with diminished overall survival and progression-free survival. Despite its clinical significance, the intricate molecular mechanisms governing this AT loss and its chemotherapy-triggered initiation remain poorly understood. Notably, the evaluation of AT remodeling in most clinical trials has predominantly relied on computerized tomography scans or bioimpedance, with molecular studies often conducted using animal or in vitro models. To address this knowledge gap, a comprehensive narrative review was conducted. The findings underscore that chemotherapy serves as a key factor in inducing AT loss, exacerbating cachexia, a paraneoplastic syndrome that significantly compromises patient quality of life and survival. The mechanism driving AT loss appears intricately linked to alterations in AT metabolic remodeling, marked by heightened lipolysis and fatty acid oxidation, coupled with diminished lipogenesis. However, adipocyte stem cells' lost ability to divide due to chemotherapy also appears to be at the root of the loss of AT. Notably, chemotherapy seems to deactivate the mitochondrial antioxidant system by reducing key regulatory enzymes responsible for neutralizing reactive oxygen species (ROS), thereby impeding lipogenesis. Despite FDG-PET evidence of AT browning, no molecular evidence of thermogenesis was reported. Prospective investigations unraveling the molecular mechanisms modulated in AT by chemotherapy, along with therapeutic strategies aimed at preventing AT loss, promise to refine treatment paradigms and enhance patient outcomes.
化疗过程中内脏和皮下脂肪组织(AT)的消耗与总生存期和无进展生存期的缩短密切相关。尽管这具有重要的临床意义,但人们对控制内脏和皮下脂肪组织减少及其化疗引发的复杂分子机制仍然知之甚少。值得注意的是,大多数临床试验中对 AT 重塑的评估主要依赖于计算机断层扫描或生物阻抗,而分子研究通常使用动物或体外模型。为了填补这一知识空白,我们进行了一项全面的叙述性综述。研究结果强调,化疗是诱发AT丧失的关键因素,会加剧恶病质,而恶病质是一种副肿瘤综合征,会严重影响患者的生活质量和生存期。促使脂肪细胞减少的机制似乎与脂肪细胞代谢重塑的改变密切相关,其特点是脂肪分解和脂肪酸氧化增加,同时脂肪生成减少。然而,化疗导致脂肪细胞干细胞丧失分裂能力,似乎也是导致脂肪细胞干细胞丧失的根本原因。值得注意的是,化疗似乎通过减少负责中和活性氧(ROS)的关键调节酶,使线粒体抗氧化系统失活,从而阻碍脂肪生成。尽管有 FDG-PET 证据表明 AT 会褐变,但没有关于产热的分子证据的报道。前瞻性研究揭示了化疗对AT的分子调控机制,以及旨在预防AT丧失的治疗策略,有望完善治疗范式并改善患者预后。
{"title":"The impact of chemotherapy on adipose tissue remodeling: The molecular players involved in this tissue wasting","authors":"Samuel Barbosa , Mafalda Barbosa Pedrosa , Rita Ferreira , Daniel Moreira-Gonçalves , Lúcio Lara Santos","doi":"10.1016/j.biochi.2024.03.016","DOIUrl":"10.1016/j.biochi.2024.03.016","url":null,"abstract":"<div><p>The depletion of visceral and subcutaneous adipose tissue (AT) during chemotherapy significantly correlates with diminished overall survival and progression-free survival. Despite its clinical significance, the intricate molecular mechanisms governing this AT loss and its chemotherapy-triggered initiation remain poorly understood. Notably, the evaluation of AT remodeling in most clinical trials has predominantly relied on computerized tomography scans or bioimpedance, with molecular studies often conducted using animal or in vitro models. To address this knowledge gap, a comprehensive narrative review was conducted. The findings underscore that chemotherapy serves as a key factor in inducing AT loss, exacerbating cachexia, a paraneoplastic syndrome that significantly compromises patient quality of life and survival. The mechanism driving AT loss appears intricately linked to alterations in AT metabolic remodeling, marked by heightened lipolysis and fatty acid oxidation, coupled with diminished lipogenesis. However, adipocyte stem cells' lost ability to divide due to chemotherapy also appears to be at the root of the loss of AT. Notably, chemotherapy seems to deactivate the mitochondrial antioxidant system by reducing key regulatory enzymes responsible for neutralizing reactive oxygen species (ROS), thereby impeding lipogenesis. Despite FDG-PET evidence of AT browning, no molecular evidence of thermogenesis was reported. Prospective investigations unraveling the molecular mechanisms modulated in AT by chemotherapy, along with therapeutic strategies aimed at preventing AT loss, promise to refine treatment paradigms and enhance patient outcomes.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0300908424000749/pdfft?md5=058b3ddc3df668c738a551bf6c562da4&pid=1-s2.0-S0300908424000749-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140308294","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The blood-brain barrier (BBB) is a protective semi-permeable structure that regulates the exchange of biomolecules between the peripheral blood and the central nervous system (CNS). Due to its specialized tight junctions and low vesicle trafficking, the BBB strictly limits the paracellular passage and transcellular transport of molecules to maintain the physiological condition of brain tissues. BBB breakdown is associated with many CNS disorders. Soluble epoxide hydrolase (sEH) is a hydrolase enzyme that converts epoxy-fatty acids (EpFAs) to their corresponding diols and is involved in the onset and progression of multiple diseases. EpFAs play a protective role in the central nervous system via preventing neuroinflammation, making sEH a potential therapeutic target for CNS diseases. Recent studies showed that sEH inhibition prevented BBB impairment caused by stroke, hemorrhage, traumatic brain injury, hyperglycemia and sepsis via regulating the expression of tight junctions. In this review, the protective actions of sEH inhibition on BBB and potential mechanisms are summarized, and some important questions that remain to be resolved are also addressed.
{"title":"Inhibition of soluble epoxide hydrolase as a therapeutic approach for blood-brain barrier dysfunction","authors":"Shuo Li, Huijia Song, Yanping Sun, Yongjun Sun, Huimin Zhang, Zibin Gao","doi":"10.1016/j.biochi.2024.03.015","DOIUrl":"10.1016/j.biochi.2024.03.015","url":null,"abstract":"<div><p>The blood-brain barrier (BBB) is a protective semi-permeable structure that regulates the exchange of biomolecules between the peripheral blood and the central nervous system (CNS). Due to its specialized tight junctions and low vesicle trafficking, the BBB strictly limits the paracellular passage and transcellular transport of molecules to maintain the physiological condition of brain tissues. BBB breakdown is associated with many CNS disorders. Soluble epoxide hydrolase (sEH) is a hydrolase enzyme that converts epoxy-fatty acids (EpFAs) to their corresponding diols and is involved in the onset and progression of multiple diseases. EpFAs play a protective role in the central nervous system via preventing neuroinflammation, making sEH a potential therapeutic target for CNS diseases. Recent studies showed that sEH inhibition prevented BBB impairment caused by stroke, hemorrhage, traumatic brain injury, hyperglycemia and sepsis via regulating the expression of tight junctions. In this review, the protective actions of sEH inhibition on BBB and potential mechanisms are summarized, and some important questions that remain to be resolved are also addressed.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140295548","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-18DOI: 10.1016/j.biochi.2024.03.010
Jean A. Boutin , Valérie Hamon de Almeida , Nathalie Coussay , Céline Legros , Gilles Ferry , Karine Reybier
Among the properties melatonin is claimed to possess, are the immuno-inflammation inductive capacities that would be responsible of some of the paramount of activities melatonin is reported to have in most of the human pathological conditions. In the present paper, we measured the effect of melatonin on established cellular models of immuno-inflammation, and found none. The discrepancies are discussed, especially because those properties are reported at pharmacological concentration (1 μM and beyond) at which the melatonin receptors are desensitized by internalization, leading to putative non-receptor-dependent mechanism of action.
{"title":"Melatonin facts: Melatonin lacks immuno-inflammation boosting capacities at the molecular and cellular levels","authors":"Jean A. Boutin , Valérie Hamon de Almeida , Nathalie Coussay , Céline Legros , Gilles Ferry , Karine Reybier","doi":"10.1016/j.biochi.2024.03.010","DOIUrl":"10.1016/j.biochi.2024.03.010","url":null,"abstract":"<div><p>Among the properties melatonin is claimed to possess, are the immuno-inflammation inductive capacities that would be responsible of some of the paramount of activities melatonin is reported to have in most of the human pathological conditions. In the present paper, we measured the effect of melatonin on established cellular models of immuno-inflammation, and found none. The discrepancies are discussed, especially because those properties are reported at pharmacological concentration (1 μM and beyond) at which the melatonin receptors are desensitized by internalization, leading to putative non-receptor-dependent mechanism of action.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.9,"publicationDate":"2024-03-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140178220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-03-16DOI: 10.1016/j.biochi.2024.03.009
Translocator protein (TSPO) is an 18 kDa transmembrane protein, localized primarily on the outer mitochondrial membrane. It has been found to be involved in various physiological processes and pathophysiological conditions. Though studies on its structure have been performed only recently, there is little information on the nature of dynamics and doubts about some structures referenced in the literature, especially the NMR structure of mouse TSPO. In the present work, we thoroughly study the dynamics of mouse TSPO protein by means of atomistic molecular dynamics simulations, in presence as well as in absence of the diagnostic ligand PKA. We considered two starting structures: the NMR structure and a homology model (HM) generated on the basis of X-ray structures from bacterial TSPO. We examine the conformational landscape in both the modes for both starting points, in presence and absence of the ligand, in order to measure its impact for both structures. The analysis highlights high flexibility of the protein globally, but NMR simulations show a surprisingly flexibility even in the presence of the ligand. Interestingly, this is not the case for HM calculations, to the point that the ligand seems not so stable as in the NMR system and an unbinding event process is partially sampled. All those results tend to show that the NMR structure of mTSPO seems not deficient but is just in another portion of the global conformation space of TSPO.
{"title":"The mutual and dynamic role of TSPO and ligands in their binding process: An example with PK-11195","authors":"","doi":"10.1016/j.biochi.2024.03.009","DOIUrl":"10.1016/j.biochi.2024.03.009","url":null,"abstract":"<div><p>Translocator protein (TSPO) is an 18 kDa transmembrane protein, localized primarily on the outer mitochondrial membrane. It has been found to be involved in various physiological processes and pathophysiological conditions. Though studies on its structure have been performed only recently, there is little information on the nature of dynamics and doubts about some structures referenced in the literature, especially the NMR structure of mouse TSPO. In the present work, we thoroughly study the dynamics of mouse TSPO protein by means of atomistic molecular dynamics simulations, in presence as well as in absence of the diagnostic ligand PKA. We considered two starting structures: the NMR structure and a homology model (HM) generated on the basis of X-ray structures from bacterial TSPO. We examine the conformational landscape in both the modes for both starting points, in presence and absence of the ligand, in order to measure its impact for both structures. The analysis highlights high flexibility of the protein globally, but NMR simulations show a surprisingly flexibility even in the presence of the ligand. Interestingly, this is not the case for HM calculations, to the point that the ligand seems not so stable as in the NMR system and an unbinding event process is partially sampled. All those results tend to show that the NMR structure of mTSPO seems not deficient but is just in another portion of the global conformation space of TSPO.</p></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2024-03-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0300908424000671/pdfft?md5=b7784e0a84b2e69202dea04a8e4b84c0&pid=1-s2.0-S0300908424000671-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140144834","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}