Pub Date : 2022-09-01DOI: 10.1080/10715762.2022.2162392
Cem Koray Firat, Beyza Nur Ozkan, Eray Metin Guler
Vitamin B12 is involved in biochemical metabolic pathways. B12 deficiency is common in childhood when the need for the vitamin increases and growth and development occur. Various hematological, neurological, psychiatric, and gastrointestinal disorders are observed in its deficiency. In addition, B12 deficiency is associated with oxidative stress and DNA damage. Therefore, the aim of our study is to evaluate oxidative stress, thiol/disulfide homeostasis, and DNA damage pre and post-treatment in children diagnosed with B12 deficiency. A total of 40 children with B12 deficiency were included in the study after the consent form was approved. Blood was drawn from children pre and posttreatment. Hemoglobin (HGB), hematocrit (HCT), and red blood cells (RBC) were measured by autoanalyzer; total antioxidant status (TAS), total oxidant status (TOS), total thiol (TT), and native thiol (NT) were measured by the photometric method, and DNA damage was analyzed by the comet assay method. Oxidative stress index (OSI) and disulfide (DIS) values were calculated. As a result of the experiments, HGB, HCT, and RBC increased with treatment. While TAS, TT, and NT as antioxidant parameters increased; TOS, OSI, and DIS decreased with treatment compared to pretreatment. DNA damage was also found to decrease with treatment. Additionally, these data were statistically significant (p < 0.001). It was found that oxidative stress and DNA damage decreased with oral B12 treatment in children with B12 deficiency, and clinical parameters were also improved.
{"title":"Beneficial effects of vitamin B<sub>12</sub> treatment in pediatric patients diagnosed with vitamin B<sub>12</sub> deficiency regarding total-native thiol, oxidative stress, and mononuclear leukocyte DNA damage.","authors":"Cem Koray Firat, Beyza Nur Ozkan, Eray Metin Guler","doi":"10.1080/10715762.2022.2162392","DOIUrl":"https://doi.org/10.1080/10715762.2022.2162392","url":null,"abstract":"<p><p>Vitamin B<sub>12</sub> is involved in biochemical metabolic pathways. B<sub>12</sub> deficiency is common in childhood when the need for the vitamin increases and growth and development occur. Various hematological, neurological, psychiatric, and gastrointestinal disorders are observed in its deficiency. In addition, B<sub>12</sub> deficiency is associated with oxidative stress and DNA damage. Therefore, the aim of our study is to evaluate oxidative stress, thiol/disulfide homeostasis, and DNA damage pre and post-treatment in children diagnosed with B<sub>12</sub> deficiency. A total of 40 children with B<sub>12</sub> deficiency were included in the study after the consent form was approved. Blood was drawn from children pre and posttreatment. Hemoglobin (HGB), hematocrit (HCT), and red blood cells (RBC) were measured by autoanalyzer; total antioxidant status (TAS), total oxidant status (TOS), total thiol (TT), and native thiol (NT) were measured by the photometric method, and DNA damage was analyzed by the comet assay method. Oxidative stress index (OSI) and disulfide (DIS) values were calculated. As a result of the experiments, HGB, HCT, and RBC increased with treatment. While TAS, TT, and NT as antioxidant parameters increased; TOS, OSI, and DIS decreased with treatment compared to pretreatment. DNA damage was also found to decrease with treatment. Additionally, these data were statistically significant (<i>p</i> < 0.001). It was found that oxidative stress and DNA damage decreased with oral B<sub>12</sub> treatment in children with B<sub>12</sub> deficiency, and clinical parameters were also improved.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10651736","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 : 2022-09-01DOI: 10.1080/10715762.2022.2162393
Lingli Lin, Xi Chen, Xiaoting Sun, Baoping Xiao, Jian Li, Jingwen Liu, Guiling Li
As a natural polyphenolic food supplement and the principal curcuminoid in turmeric, curcumin shows antioxidant, anti-inflammatory, and antitumor activities. However, its specific functional mechanism remains unclear. Our preliminary study indicated that miR-125b-5p was downregulated by a curcumin extract. This study aimed to determine whether miR-125b-5p is involved in the antioxidant regulation of curcumin. The results showed that miR-125b-5p overexpression had a pro-oxidant effect by reducing the cellular antioxidant capacity, as well as decreasing the activities of catalase (CAT) and superoxide dismutase (SOD) in the normal liver cell line LO2. However, miR-125b-5p repression significantly increased the cellular antioxidant capacity and enhanced the activities of CAT and SOD. Further investigation demonstrated that the cellular antioxidant capacity induced by curcumin extract was inhibited by miR-125b-5p overexpression. Thus, curcumin may exhibit antioxidant effects by repressing miR-125b-5p expression, which provides new insights into the molecular antioxidant mechanism of curcumin and other functional food components.
{"title":"MiR-125b-5p is targeted by curcumin to regulate the cellular antioxidant capacity.","authors":"Lingli Lin, Xi Chen, Xiaoting Sun, Baoping Xiao, Jian Li, Jingwen Liu, Guiling Li","doi":"10.1080/10715762.2022.2162393","DOIUrl":"https://doi.org/10.1080/10715762.2022.2162393","url":null,"abstract":"<p><p>As a natural polyphenolic food supplement and the principal curcuminoid in turmeric, curcumin shows antioxidant, anti-inflammatory, and antitumor activities. However, its specific functional mechanism remains unclear. Our preliminary study indicated that miR-125b-5p was downregulated by a curcumin extract. This study aimed to determine whether miR-125b-5p is involved in the antioxidant regulation of curcumin. The results showed that miR-125b-5p overexpression had a pro-oxidant effect by reducing the cellular antioxidant capacity, as well as decreasing the activities of catalase (CAT) and superoxide dismutase (SOD) in the normal liver cell line LO2. However, miR-125b-5p repression significantly increased the cellular antioxidant capacity and enhanced the activities of CAT and SOD. Further investigation demonstrated that the cellular antioxidant capacity induced by curcumin extract was inhibited by miR-125b-5p overexpression. Thus, curcumin may exhibit antioxidant effects by repressing miR-125b-5p expression, which provides new insights into the molecular antioxidant mechanism of curcumin and other functional food components.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9204253","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 : 2022-09-01DOI: 10.1080/10715762.2022.2157272
Yasumasa Okazaki, Nanami Ito, Hiromasa Tanaka, Masaru Hori, Shinya Toyokuni
Non-thermal plasma (NTP) induces the generation of reactive oxygen species (ROS) and reactive nitrogen species, such as hydroxyl radicals (•OH), hydrogen peroxide (H2O2), singlet oxygen, superoxide, ozone, and nitric oxide, at near-physiological temperatures. These molecules promote blood coagulation, wound healing, disinfection, and selective cancer cell death. Based on these evidences, clinical trials of NTP have been conducted for treating chronic wounds and head and neck cancers. Although clinical applications have progressed, the stoichiometric quantification of NTP-induced ROS remains unclear in the liquid phase in the presence of FeCl2 or FeCl3 in combination with biocompatible reducing agents, which may modulate the final biological effects of NTP. In this study, we employed electron paramagnetic resonance spectroscopy to quantify ROS using spin-trapping probe, 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and H2O2, using luminescent probe in the presence of FeCl2 or FeCl3. NTP-induced DMPO-OH levels were elevated 10-100 µM FeCl2 or 500 and 1000 µM FeCl3. NTP-induced DMPO-OH with 10 µM FeCl2 or FeCl3 was significantly scavenged by ascorbate, α-tocopherol, dithiothreitol, reduced glutathione, or oxidized glutathione, whereas dehydroascorbate was ineffective in 2 mM DMPO. NTP-induced H2O2 was significantly degraded by 100 µM FeCl2 and FeCl3 in an iron-dependent manner. Meanwhile, decomposition of H2O2 by catalase decayed DMPO-OH efficiently in the presence of iron, indicating iron causes DMPO-OH production and degradation simultaneously. These results suggest that NTP-induced DMPO-OH is generated by the H2O2-consuming, iron-dependent Fenton reaction and ferryl intermediates. The potential iron-mediated ROS production by NTP is also discussed to clarify the interaction between NTP-induced ROS and biomolecules.
非热等离子体(NTP)在接近生理温度下诱导活性氧(ROS)和活性氮的产生,如羟基自由基(•OH)、过氧化氢(H2O2)、单线态氧、超氧化物、臭氧和一氧化氮。这些分子促进血液凝固、伤口愈合、消毒和选择性癌细胞死亡。基于这些证据,已经开展了治疗慢性伤口和头颈部癌症的NTP临床试验。尽管临床应用已取得进展,但在FeCl2或FeCl3与生物相容性还原剂联合存在的液相中,NTP诱导的ROS的化学计量定量尚不清楚,这可能会调节NTP的最终生物学效应。在本研究中,我们采用电子顺磁共振波谱法,在FeCl2或FeCl3存在下,使用自旋捕获探针,5,5-二甲基-1-吡咯啉- n-氧化物(DMPO)和H2O2,使用发光探针来量化ROS。ntp诱导的dpo - oh水平升高10-100µM FeCl2或500和1000µM FeCl3。抗坏血酸、α-生育酚、二硫苏糖醇、还原性谷胱甘肽或氧化性谷胱甘肽均能清除10 μ M FeCl2或FeCl3诱导的ntp诱导的dpo - oh,而脱氢抗坏血酸在2 mM DMPO中无效。ntp诱导的H2O2被100µM FeCl2和FeCl3以铁依赖的方式显著降解。同时,过氧化氢酶对H2O2的分解在铁存在的情况下能有效地降解dpo - oh,说明铁同时导致dpo - oh的产生和降解。这些结果表明,ntp诱导的DMPO-OH是由消耗h2o2、依赖铁的Fenton反应和铁基中间体产生的。为了阐明NTP诱导的ROS与生物分子之间的相互作用,我们还讨论了NTP潜在的铁介导ROS的产生。
{"title":"Non-thermal plasma elicits ferrous chloride-catalyzed DMPO-OH.","authors":"Yasumasa Okazaki, Nanami Ito, Hiromasa Tanaka, Masaru Hori, Shinya Toyokuni","doi":"10.1080/10715762.2022.2157272","DOIUrl":"https://doi.org/10.1080/10715762.2022.2157272","url":null,"abstract":"<p><p>Non-thermal plasma (NTP) induces the generation of reactive oxygen species (ROS) and reactive nitrogen species, such as hydroxyl radicals (<sup>•</sup>OH), hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), singlet oxygen, superoxide, ozone, and nitric oxide, at near-physiological temperatures. These molecules promote blood coagulation, wound healing, disinfection, and selective cancer cell death. Based on these evidences, clinical trials of NTP have been conducted for treating chronic wounds and head and neck cancers. Although clinical applications have progressed, the stoichiometric quantification of NTP-induced ROS remains unclear in the liquid phase in the presence of FeCl<sub>2</sub> or FeCl<sub>3</sub> in combination with biocompatible reducing agents, which may modulate the final biological effects of NTP. In this study, we employed electron paramagnetic resonance spectroscopy to quantify ROS using spin-trapping probe, 5,5-dimethyl-1-pyrroline-<i>N</i>-oxide (DMPO) and H<sub>2</sub>O<sub>2</sub>, using luminescent probe in the presence of FeCl<sub>2</sub> or FeCl<sub>3</sub>. NTP-induced DMPO-OH levels were elevated 10-100 µM FeCl<sub>2</sub> or 500 and 1000 µM FeCl<sub>3</sub>. NTP-induced DMPO-OH with 10 µM FeCl<sub>2</sub> or FeCl<sub>3</sub> was significantly scavenged by ascorbate, α-tocopherol, dithiothreitol, reduced glutathione, or oxidized glutathione, whereas dehydroascorbate was ineffective in 2 mM DMPO. NTP-induced H<sub>2</sub>O<sub>2</sub> was significantly degraded by 100 µM FeCl<sub>2</sub> and FeCl<sub>3</sub> in an iron-dependent manner. Meanwhile, decomposition of H<sub>2</sub>O<sub>2</sub> by catalase decayed DMPO-OH efficiently in the presence of iron, indicating iron causes DMPO-OH production and degradation simultaneously. These results suggest that NTP-induced DMPO-OH is generated by the H<sub>2</sub>O<sub>2</sub>-consuming, iron-dependent Fenton reaction and ferryl intermediates. The potential iron-mediated ROS production by NTP is also discussed to clarify the interaction between NTP-induced ROS and biomolecules.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10651269","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 : 2022-09-01DOI: 10.1080/10715762.2022.2163244
Yangying Li, Yuda Zhu, Shiyi Li, Yuman Dong, Chengyu Wan, Xiuxian Yu, Guang Xin, Zeliang Wei, Fan Li, Yilan Wang, Kun Zhang, Qingqiu Chen, Cuicui Zhang, E Wen, Hai Niu, Wen Huang
Severe acute pancreatitis (SAP) is an inflammatory disorder of the exocrine pancreas associated with high morbidity and mortality. SAP has been proven to trigger mitochondria dysfunction in the pancreas. We found that Deoxyarbutin (dA) recovered impaired mitochondrial function. High-temperature requirement protein A2 (HtrA2), a mitochondrial serine protease upstream of PGC-1α, is charge of quality control in mitochondrial homeostasis. The molecular docking study indicated that there was a potential interaction between dA and HtrA2. However, whether the protective effect of dA against SAP is regulated by HtrA2/PGC-1α remains unknown. Our study in vitro showed that dA significantly reduced the necrosis of primary acinar cells and reactive oxygen species (ROS) accumulation, recovered mitochondrial membrane potential (ΔΨm) and ATP exhaustion, while UCF-101 (HtrA2 inhibitor), and SR-18292 (PGC-1α inhibitor) eliminated the protective effect of dA. Moreover, HtrA2 siRNA transfection efficiently blocked the protective of dA on HtrA2/PGC-1α pathway in 266-6 acinar cells. Meanwhile, dA also decreased LC3II/I ration, as well as p62, and increased Parkin expression, while UCF-101 and Bafilomycin A1 (autophagy inhibitor) reversed the protective effect of dA. Our study in vivo confirmed that dA effectively alleviated severity of SAP by reducing pancreatic edema, plasma amylase, and lipase levels and improved the HtrA2/PGC-1α pathway. Therefore, this is the first study to identify that dA inhibits pancreatic injury caused by oxidative stress, mitochondrial dysfunction, and impaired autophagy in a HtrA2/PGC-1α dependent manner.
{"title":"Deoxyarbutin attenuates severe acute pancreatitis via the HtrA2/PGC-1α pathway.","authors":"Yangying Li, Yuda Zhu, Shiyi Li, Yuman Dong, Chengyu Wan, Xiuxian Yu, Guang Xin, Zeliang Wei, Fan Li, Yilan Wang, Kun Zhang, Qingqiu Chen, Cuicui Zhang, E Wen, Hai Niu, Wen Huang","doi":"10.1080/10715762.2022.2163244","DOIUrl":"https://doi.org/10.1080/10715762.2022.2163244","url":null,"abstract":"<p><p>Severe acute pancreatitis (SAP) is an inflammatory disorder of the exocrine pancreas associated with high morbidity and mortality. SAP has been proven to trigger mitochondria dysfunction in the pancreas. We found that Deoxyarbutin (dA) recovered impaired mitochondrial function. High-temperature requirement protein A2 (HtrA2), a mitochondrial serine protease upstream of PGC-1α, is charge of quality control in mitochondrial homeostasis. The molecular docking study indicated that there was a potential interaction between dA and HtrA2. However, whether the protective effect of dA against SAP is regulated by HtrA2/PGC-1α remains unknown. Our study <i>in vitro</i> showed that dA significantly reduced the necrosis of primary acinar cells and reactive oxygen species (ROS) accumulation, recovered mitochondrial membrane potential (ΔΨm) and ATP exhaustion, while UCF-101 (HtrA2 inhibitor), and SR-18292 (PGC-1α inhibitor) eliminated the protective effect of dA. Moreover, HtrA2 siRNA transfection efficiently blocked the protective of dA on HtrA2/PGC-1α pathway in 266-6 acinar cells. Meanwhile, dA also decreased LC3II/I ration, as well as p62, and increased Parkin expression, while UCF-101 and Bafilomycin A1 (autophagy inhibitor) reversed the protective effect of dA. Our study <i>in vivo</i> confirmed that dA effectively alleviated severity of SAP by reducing pancreatic edema, plasma amylase, and lipase levels and improved the HtrA2/PGC-1α pathway. Therefore, this is the first study to identify that dA inhibits pancreatic injury caused by oxidative stress, mitochondrial dysfunction, and impaired autophagy in a HtrA2/PGC-1α dependent manner.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10651745","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}
Hydrogen sulfide, hydropersulfides, and hydropolysulfides have been revealed to play important physiological roles such as cell signaling and protection against oxidative stress, but the underlying mechanisms and dynamics of action remain elusive. It is generally accepted that these species act by two-electron redox mechanisms, while the involvement of one-electron redox chemistry has received less attention. In this study, the radical-scavenging activity of hydrogen persulfide, hydrogen polysulfides (HSnH n = 2-4), and diallyl- or dialkyl-sulfides (RSnR, n = 1-4) was measured. Furthermore, their antioxidant effects against free radical-mediated human plasma lipid peroxidation were assessed by measuring lipid hydroperoxides. It was found that disodium disulfide, trisulfide, and tetrasulfide acted as potent peroxyl radical scavengers, the rate constant for scavenging peroxyl radical being 3.5 × 105, 4.0 × 105, and 6.0 × 105 M-1 s-1 in PBS pH 7.4 at 37 °C respectively and that they inhibited plasma lipid peroxidation efficiently, the efficacy is increased with the catenation number. Disodium tetrasulfide was 1.5 times as reactive as Trolox toward peroxyl radical and inhibited plasma lipid peroxidation more efficiently than ascorbate and Trolox. On the other hand, diallyl- and dialkyl-sulfides did not exert significant radical-scavenging activity, nor did they inhibit lipid peroxidation efficiently, except for diallyl tetrasulfide, which suppressed plasma lipid peroxidation, despite less significantly than disodium tetrasulfide. Collectively, this study shows that hydrogen persulfide and hydrogen polysulfides act as potent radical-scavenging antioxidants and that, in addition to two-electron redox mechanisms, one electron redox reaction may also play important role in the in vivo defense against deleterious oxidative stress.
硫化氢、氢过硫化物和氢多硫化物已被发现在细胞信号传导和抗氧化应激方面发挥重要的生理作用,但其潜在的机制和作用动力学尚不清楚。人们普遍认为这些物质是通过双电子氧化还原机制起作用的,而单电子氧化还原化学的参与却很少受到关注。本研究测定了过硫化氢、多硫化氢(HSnH n = 2-4)和二烯丙基或二烷基硫化物(RSnR, n = 1-4)的自由基清除活性。此外,通过测量脂质氢过氧化物来评估它们对自由基介导的人血浆脂质过氧化的抗氧化作用。结果表明,二硫二钠、三硫二钠和四硫二钠具有较强的过氧自由基清除作用,在37℃、pH为7.4的PBS条件下,对过氧自由基的清除速率分别为3.5 × 105、4.0 × 105和6.0 × 105 M-1 s-1,且能有效抑制血脂过氧化,其作用随阳离子化数的增加而增强。四硫二钠对过氧自由基的活性是Trolox的1.5倍,并且比抗坏血酸盐和Trolox更有效地抑制血浆脂质过氧化。另一方面,二烯丙基和二烷基硫化物没有发挥显著的自由基清除活性,也没有有效地抑制脂质过氧化,除了二烯丙基四硫醚抑制血浆脂质过氧化,尽管不如四硫二钠显著。总之,本研究表明,过硫化氢和多硫化氢是有效的自由基清除抗氧化剂,除了双电子氧化还原机制外,单电子氧化还原反应也可能在体内防御有害氧化应激中发挥重要作用。
{"title":"Antioxidant action of persulfides and polysulfides against free radical-mediated lipid peroxidation.","authors":"Takayuki Kaneko, Yuichiro Mita, Kanako Nozawa-Kumada, Masana Yazaki, Mieko Arisawa, Etsuo Niki, Noriko Noguchi, Yoshiro Saito","doi":"10.1080/10715762.2023.2165918","DOIUrl":"https://doi.org/10.1080/10715762.2023.2165918","url":null,"abstract":"<p><p>Hydrogen sulfide, hydropersulfides, and hydropolysulfides have been revealed to play important physiological roles such as cell signaling and protection against oxidative stress, but the underlying mechanisms and dynamics of action remain elusive. It is generally accepted that these species act by two-electron redox mechanisms, while the involvement of one-electron redox chemistry has received less attention. In this study, the radical-scavenging activity of hydrogen persulfide, hydrogen polysulfides (HS<sub>n</sub>H <i>n</i> = 2-4), and diallyl- or dialkyl-sulfides (RS<sub>n</sub>R, <i>n</i> = 1-4) was measured. Furthermore, their antioxidant effects against free radical-mediated human plasma lipid peroxidation were assessed by measuring lipid hydroperoxides. It was found that disodium disulfide, trisulfide, and tetrasulfide acted as potent peroxyl radical scavengers, the rate constant for scavenging peroxyl radical being 3.5 × 10<sup>5</sup>, 4.0 × 10<sup>5</sup>, and 6.0 × 10<sup>5</sup> M<sup>-1</sup> s<sup>-1</sup> in PBS pH 7.4 at 37 °C respectively and that they inhibited plasma lipid peroxidation efficiently, the efficacy is increased with the catenation number. Disodium tetrasulfide was 1.5 times as reactive as Trolox toward peroxyl radical and inhibited plasma lipid peroxidation more efficiently than ascorbate and Trolox. On the other hand, diallyl- and dialkyl-sulfides did not exert significant radical-scavenging activity, nor did they inhibit lipid peroxidation efficiently, except for diallyl tetrasulfide, which suppressed plasma lipid peroxidation, despite less significantly than disodium tetrasulfide. Collectively, this study shows that hydrogen persulfide and hydrogen polysulfides act as potent radical-scavenging antioxidants and that, in addition to two-electron redox mechanisms, one electron redox reaction may also play important role in the <i>in vivo</i> defense against deleterious oxidative stress.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9219837","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}
3,4-Dihydroxyphenylacetic acid (DOPAC) and 3-hydroxyphenylacetic acid (OPAC) are the predominant catabolites of quercetin glycosides, such as quercetin 4'-O-β-glucoside from the onion, produced by intestinal microbiota. Although each catabolite has been reported to protect the cells from acetaldehyde-induced cytotoxicity, the effect of their combination remains to be clarified. The purpose of this study was to determine whether the combination of DOPAC and OPAC enhances the resistance against the acetaldehyde-induced oxidative stress in the cultured hepatocytes. The pretreatment of the combination of DOPAC (5 μM) and OPAC (5 μM) showed significant protection against the acetaldehyde- and hydrogen peroxide-induced cytotoxicity, even though each compound at the same concentration did not. This combination also significantly inhibited the intracellular dichlorofluorescin diacetate-detectable reactive oxygen species (ROS) level, whereas the solo treatment did slightly, suggesting that reducing mechanisms of ROS or compounds that enhance ROS production are involved in the cytoprotective effect. The combinatory treatment significantly enhanced the gene expression of not only the aldehyde dehydrogenases (ALDHs), but also glutamate-cysteine ligase, catalytic subunit, the first rate-limiting enzyme of glutathione (GSH) synthesis. Accordingly, both the intracellular GSH level and the total ALDH activity were enhanced by DOPAC plus OPAC. Involvement of GSH in the cytoprotection as well as ALDH up-regulation by the combination was confirmed by the experiments using a GSH biosynthesis inhibitor, buthionine sulfoximine. Taken together, the present results suggested that the quercetin microbiota catabolites concertedly protect the cells from acetaldehyde through a pre-enhanced resistance against oxidative stress by the GSH-dependent up-regulation of ALDHs.
{"title":"The microbiota catabolites of quercetin glycosides concertedly enhance the resistance against acetaldehyde-induced oxidative stress.","authors":"Kexin Li, Hongyan Wu, Minori Kidawara, Yun Lin, Ayano Satoh, Gongliang Zhang, Shintaro Munemasa, Yoshiyuki Murata, Toshiyuki Nakamura, Yoshimasa Nakamura","doi":"10.1080/10715762.2022.2159820","DOIUrl":"https://doi.org/10.1080/10715762.2022.2159820","url":null,"abstract":"<p><p>3,4-Dihydroxyphenylacetic acid (DOPAC) and 3-hydroxyphenylacetic acid (OPAC) are the predominant catabolites of quercetin glycosides, such as quercetin 4'-<i>O</i>-β-glucoside from the onion, produced by intestinal microbiota. Although each catabolite has been reported to protect the cells from acetaldehyde-induced cytotoxicity, the effect of their combination remains to be clarified. The purpose of this study was to determine whether the combination of DOPAC and OPAC enhances the resistance against the acetaldehyde-induced oxidative stress in the cultured hepatocytes. The pretreatment of the combination of DOPAC (5 μM) and OPAC (5 μM) showed significant protection against the acetaldehyde- and hydrogen peroxide-induced cytotoxicity, even though each compound at the same concentration did not. This combination also significantly inhibited the intracellular dichlorofluorescin diacetate-detectable reactive oxygen species (ROS) level, whereas the solo treatment did slightly, suggesting that reducing mechanisms of ROS or compounds that enhance ROS production are involved in the cytoprotective effect. The combinatory treatment significantly enhanced the gene expression of not only the aldehyde dehydrogenases (ALDHs), but also glutamate-cysteine ligase, catalytic subunit, the first rate-limiting enzyme of glutathione (GSH) synthesis. Accordingly, both the intracellular GSH level and the total ALDH activity were enhanced by DOPAC plus OPAC. Involvement of GSH in the cytoprotection as well as ALDH up-regulation by the combination was confirmed by the experiments using a GSH biosynthesis inhibitor, buthionine sulfoximine. Taken together, the present results suggested that the quercetin microbiota catabolites concertedly protect the cells from acetaldehyde through a pre-enhanced resistance against oxidative stress by the GSH-dependent up-regulation of ALDHs.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10649535","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}
Drug repurposing allows searching for new biological targets, especially against emerging diseases such as Covid-19. Drug colchicine (COL) presents recognized anti-inflammatory action, while the nanotechnology purpose therapies with low doses, efficacy, and decrease the drug's side-effects. This study aims to evaluate the effects of COL and colchicine nanocapsules (NCCOL) on survival, LC50, activity locomotor, and oxidative stress parameters, elucidating the toxicity profile in acute and chronic exposure in Drosophila melanogaster. Three-day-old flies were investigated into groups: Control, 0.001, 0.0025, 0.005, and 0.010 mg/mL of COL or NCCOL. The survival rate, open field test, LC50, oxidative stress markers (reactive species (RS) production, thiobarbituric acid reactive substances), antioxidant enzyme activity (catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase), protein thiols, nonprotein thiols, acetylcholinesterase activity, and cell viability were measured. As a result, acute exposure to the COL decreases the number of crosses in the open field and increases CAT activity. NCCOL reduced RS levels, increased lipoperoxidation and SOD activity. Chronic exposure to the COL and NCCOL in high concentrations implied high mortality and enzymatic inhibition of the CAT and AChE, and only the COL caused locomotor damage in the open field test. Thus, NCCOL again reduced the formation of RS while COL increased. In this comparative study, NCCOL was less toxic to the antioxidant system than COL and showed notable involvement of oxidative stress as one of their toxicity mechanisms. Future studies are needed to elucidate all aspects of nanosafety related to the NCCOL.
{"title":"Relationship between toxicity and oxidative stress of the nanoencapsulated colchicine in a model of <i>Drosophila melanogaster</i>.","authors":"Franciéle Romero Machado, Stífani Machado Araujo, Ana Cláudia Ribeiro Funguetto, Vandreza Cardoso Bortolotto, Eliana Jardim Fernandes, Munir Mustafa Dahleh Mustafa, Sandra Elisa Haas, Gustavo Petri Guerra, Marina Prigol, Silvana Peterini Boeira","doi":"10.1080/10715762.2022.2146500","DOIUrl":"https://doi.org/10.1080/10715762.2022.2146500","url":null,"abstract":"<p><p>Drug repurposing allows searching for new biological targets, especially against emerging diseases such as Covid-19. Drug colchicine (COL) presents recognized anti-inflammatory action, while the nanotechnology purpose therapies with low doses, efficacy, and decrease the drug's side-effects. This study aims to evaluate the effects of COL and colchicine nanocapsules (NCCOL) on survival, LC50, activity locomotor, and oxidative stress parameters, elucidating the toxicity profile in acute and chronic exposure in <i>Drosophila melanogaster</i>. Three-day-old flies were investigated into groups: Control, 0.001, 0.0025, 0.005, and 0.010 mg/mL of COL or NCCOL. The survival rate, open field test, LC50, oxidative stress markers (reactive species (RS) production, thiobarbituric acid reactive substances), antioxidant enzyme activity (catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase), protein thiols, nonprotein thiols, acetylcholinesterase activity, and cell viability were measured. As a result, acute exposure to the COL decreases the number of crosses in the open field and increases CAT activity. NCCOL reduced RS levels, increased lipoperoxidation and SOD activity. Chronic exposure to the COL and NCCOL in high concentrations implied high mortality and enzymatic inhibition of the CAT and AChE, and only the COL caused locomotor damage in the open field test. Thus, NCCOL again reduced the formation of RS while COL increased. In this comparative study, NCCOL was less toxic to the antioxidant system than COL and showed notable involvement of oxidative stress as one of their toxicity mechanisms. Future studies are needed to elucidate all aspects of nanosafety related to the NCCOL.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10657696","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 : 2022-09-01DOI: 10.1080/10715762.2022.2161379
Aiswarya Purushothaman, Smrithi S Babu, Surya Naroth, Deepa Janardanan
Caffeic acid is a phenolic secondary metabolite from plants, which is known for its antioxidant properties. The effective mitigation of methanol-induced oxidative stress by caffeic acid depends on the direct radical scavenging as well as the formation of new metabolites via oxidative degradation. Herein, thermodynamic and kinetic aspects of the oxidative degradation pathway of caffeic acid in the presence of radical CH3O• and its isomer, •CH2OH are discussed for the first time, employing density functional theory (DFT). The direct radical scavenging activity of caffeic acid against these radicals is verified via hydrogen atom transfer (HAT) and radical adduct formation (RAF) mechanisms. HAT is predicted to be more feasible than RAF mechanism as per the computed data. Additionally, energetic details of the proposed oxidative degradation pathway of radical adduct intermediates toward the formation of a cyclic metabolite is analyzed. Kinetic studies indicated a significant tunneling contribution to the H abstraction pathways having high activation barriers. Further, our results imply that the newly formed metabolites exhibit comparable antioxidant activity with that of caffeic acid.
{"title":"Antioxidant activity of caffeic acid: thermodynamic and kinetic aspects on the oxidative degradation pathway.","authors":"Aiswarya Purushothaman, Smrithi S Babu, Surya Naroth, Deepa Janardanan","doi":"10.1080/10715762.2022.2161379","DOIUrl":"https://doi.org/10.1080/10715762.2022.2161379","url":null,"abstract":"<p><p>Caffeic acid is a phenolic secondary metabolite from plants, which is known for its antioxidant properties. The effective mitigation of methanol-induced oxidative stress by caffeic acid depends on the direct radical scavenging as well as the formation of new metabolites <i>via</i> oxidative degradation. Herein, thermodynamic and kinetic aspects of the oxidative degradation pathway of caffeic acid in the presence of radical CH<sub>3</sub>O<sup>•</sup> and its isomer, <sup>•</sup>CH<sub>2</sub>OH are discussed for the first time, employing density functional theory (DFT). The direct radical scavenging activity of caffeic acid against these radicals is verified <i>via</i> hydrogen atom transfer (HAT) and radical adduct formation (RAF) mechanisms. HAT is predicted to be more feasible than RAF mechanism as per the computed data. Additionally, energetic details of the proposed oxidative degradation pathway of radical adduct intermediates toward the formation of a cyclic metabolite is analyzed. Kinetic studies indicated a significant tunneling contribution to the H abstraction pathways having high activation barriers. Further, our results imply that the newly formed metabolites exhibit comparable antioxidant activity with that of caffeic acid.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10649120","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 : 2022-09-01DOI: 10.1080/10715762.2023.2165073
Megan F Taylor, Michael A Black, Mark B Hampton, Elizabeth C Ledgerwood
Hydrogen peroxide (H2O2) is a ubiquitous oxidant produced in a regulated manner by various enzymes in mammalian cells. H2O2 reversibly oxidizes thiol groups of cysteine residues to mediate intracellular signaling. While examples of H2O2-dependent signaling have been reported, the exact molecular mechanism(s) of signaling and the pathways affected are not well understood. Here, the transcriptomic response of Jurkat T cells to H2O2 was investigated to determine global effects on gene expression. With a low H2O2 concentration (10 µM) that did not induce an oxidative stress response or cell death, extensive changes in gene expression occurred after 4 h (6803 differentially expressed genes). Of the genes with a greater then 2-fold change in expression, 85% were upregulated suggesting that in a physiological setting H2O2 predominantly activates gene expression. Pathway analysis identified gene expression signatures associated with FOXO and NTRK signaling. These signatures were associated with an overlapping set of transcriptional regulators. Overall, our results provide a snapshot of gene expression changes in response to H2O2, which, along with further studies, will lead to new insights into the specific pathways that are activated in response to endogenous production of H2O2, and the molecular mechanisms of H2O2 signaling.
{"title":"Insights into H<sub>2</sub>O<sub>2</sub>-induced signaling in Jurkat cells from analysis of gene expression.","authors":"Megan F Taylor, Michael A Black, Mark B Hampton, Elizabeth C Ledgerwood","doi":"10.1080/10715762.2023.2165073","DOIUrl":"https://doi.org/10.1080/10715762.2023.2165073","url":null,"abstract":"<p><p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is a ubiquitous oxidant produced in a regulated manner by various enzymes in mammalian cells. H<sub>2</sub>O<sub>2</sub> reversibly oxidizes thiol groups of cysteine residues to mediate intracellular signaling. While examples of H<sub>2</sub>O<sub>2-</sub>dependent signaling have been reported, the exact molecular mechanism(s) of signaling and the pathways affected are not well understood. Here, the transcriptomic response of Jurkat T cells to H<sub>2</sub>O<sub>2</sub> was investigated to determine global effects on gene expression. With a low H<sub>2</sub>O<sub>2</sub> concentration (10 µM) that did not induce an oxidative stress response or cell death, extensive changes in gene expression occurred after 4 h (6803 differentially expressed genes). Of the genes with a greater then 2-fold change in expression, 85% were upregulated suggesting that in a physiological setting H<sub>2</sub>O<sub>2</sub> predominantly activates gene expression. Pathway analysis identified gene expression signatures associated with FOXO and NTRK signaling. These signatures were associated with an overlapping set of transcriptional regulators. Overall, our results provide a snapshot of gene expression changes in response to H<sub>2</sub>O<sub>2,</sub> which, along with further studies, will lead to new insights into the specific pathways that are activated in response to endogenous production of H<sub>2</sub>O<sub>2</sub>, and the molecular mechanisms of H<sub>2</sub>O<sub>2</sub> signaling.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9219834","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 : 2022-07-01DOI: 10.1080/10715762.2022.2146501
Maria Tolomeo, Guglielmina Chimienti, Martina Lanza, Roberto Barbaro, Alessia Nisco, Tiziana Latronico, Piero Leone, Giuseppe Petrosillo, Grazia Maria Liuzzi, Bryony Ryder, Michal Inbar-Feigenberg, Matilde Colella, Angela M S Lezza, Rikke K J Olsen, Maria Barile
Flavin adenine dinucleotide (FAD) synthase (EC 2.7.7.2), encoded by human flavin adenine dinucleotide synthetase 1 (FLAD1), catalyzes the last step of the pathway converting riboflavin (Rf) into FAD. FLAD1 variations were identified as a cause of LSMFLAD (lipid storage myopathy due to FAD synthase deficiency, OMIM #255100), resembling Multiple Acyl-CoA Dehydrogenase Deficiency, sometimes treatable with high doses of Rf; no alternative therapeutic strategies are available. We describe here cell morphological and mitochondrial alterations in dermal fibroblasts derived from a LSMFLAD patient carrying a homozygous truncating FLAD1 variant (c.745C > T) in exon 2. Despite a severe decrease in FAD synthesis rate, the patient had decreased cellular levels of Rf and flavin mononucleotide and responded to Rf treatment. We hypothesized that disturbed flavin homeostasis and Rf-responsiveness could be due to a secondary impairment in the expression of the Rf transporter 2 (RFVT2), encoded by SLC52A2, in the frame of an adaptive retrograde signaling to mitochondrial dysfunction. Interestingly, an antioxidant response element (ARE) is found in the region upstream of the transcriptional start site of SLC52A2. Accordingly, we found that abnormal mitochondrial morphology and impairments in bioenergetics were accompanied by increased cellular reactive oxygen species content and mtDNA oxidative damage. Concomitantly, an active response to mitochondrial stress is suggested by increased levels of PPARγ-co-activator-1α and Peroxiredoxin III. In this scenario, the treatment with high doses of Rf might compensate for the secondary RFVT2 molecular defect, providing a molecular rationale for the Rf responsiveness in patients with loss of function variants in FLAD1 exon 2.HIGHLIGHTSFAD synthase deficiency alters mitochondrial morphology and bioenergetics;FAD synthase deficiency triggers a mitochondrial retrograde response;FAD synthase deficiency evokes nuclear signals that adapt the expression of RFVT2.
{"title":"Retrograde response to mitochondrial dysfunctions associated to LOF variations in <i>FLAD1</i> exon 2: unraveling the importance of RFVT2.","authors":"Maria Tolomeo, Guglielmina Chimienti, Martina Lanza, Roberto Barbaro, Alessia Nisco, Tiziana Latronico, Piero Leone, Giuseppe Petrosillo, Grazia Maria Liuzzi, Bryony Ryder, Michal Inbar-Feigenberg, Matilde Colella, Angela M S Lezza, Rikke K J Olsen, Maria Barile","doi":"10.1080/10715762.2022.2146501","DOIUrl":"https://doi.org/10.1080/10715762.2022.2146501","url":null,"abstract":"<p><p>Flavin adenine dinucleotide (FAD) synthase (EC 2.7.7.2), encoded by human flavin adenine dinucleotide synthetase 1 (<i>FLAD1</i>), catalyzes the last step of the pathway converting riboflavin (Rf) into FAD. <i>FLAD1</i> variations were identified as a cause of LSMFLAD (lipid storage myopathy due to FAD synthase deficiency, OMIM #255100), resembling Multiple Acyl-CoA Dehydrogenase Deficiency, sometimes treatable with high doses of Rf; no alternative therapeutic strategies are available. We describe here cell morphological and mitochondrial alterations in dermal fibroblasts derived from a LSMFLAD patient carrying a homozygous truncating <i>FLAD1</i> variant (c.745C > T) in exon 2. Despite a severe decrease in FAD synthesis rate, the patient had decreased cellular levels of Rf and flavin mononucleotide and responded to Rf treatment. We hypothesized that disturbed flavin homeostasis and Rf-responsiveness could be due to a secondary impairment in the expression of the Rf transporter 2 (RFVT2), encoded by <i>SLC52A2,</i> in the frame of an adaptive retrograde signaling to mitochondrial dysfunction. Interestingly, an antioxidant response element (ARE) is found in the region upstream of the transcriptional start site of <i>SLC52A2</i>. Accordingly, we found that abnormal mitochondrial morphology and impairments in bioenergetics were accompanied by increased cellular reactive oxygen species content and mtDNA oxidative damage. Concomitantly, an active response to mitochondrial stress is suggested by increased levels of PPARγ-co-activator-1α and Peroxiredoxin III. In this scenario, the treatment with high doses of Rf might compensate for the secondary RFVT2 molecular defect, providing a molecular rationale for the Rf responsiveness in patients with loss of function variants in <i>FLAD1</i> exon 2.HIGHLIGHTSFAD synthase deficiency alters mitochondrial morphology and bioenergetics;FAD synthase deficiency triggers a mitochondrial retrograde response;FAD synthase deficiency evokes nuclear signals that adapt the expression of RFVT2.</p>","PeriodicalId":12411,"journal":{"name":"Free Radical Research","volume":null,"pages":null},"PeriodicalIF":3.3,"publicationDate":"2022-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9203774","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}