Lael Ceriani, Daniel E Newmire, Xavier F Gonzales, Jean Sparks, Jose Guardiola, Felix O Omoruyi
{"title":"营养剥夺对正常和2型糖尿病人骨骼肌成肌细胞氧化应激、炎症和转录组标志物的影响","authors":"Lael Ceriani, Daniel E Newmire, Xavier F Gonzales, Jean Sparks, Jose Guardiola, Felix O Omoruyi","doi":"10.1155/jnme/6661176","DOIUrl":null,"url":null,"abstract":"<p><p><b>Background:</b> Intermittent fasting has become a new fad diet that may promote an environment to facilitate muscle atrophy, placing aging, and diabetic populations at risk for muscle loss due to nutrient deprivation. The purpose of this study was to investigate how nutrient availability and serum environment influence Type 2 diabetic myoblast density and viability, autophagy-related oxidative and inflammatory markers, and upstream gene expression signaling relevant to proteostasis. <b>Methods:</b> To explore this outcome in human skeletal muscle myoblast (HSMM) and diabetic human skeletal muscle myoblast (D-HSMM), myoblast lines were cultured per standard protocol and were incubated for 12 or 24 h with either human serum (HS) or fetal bovine serum (FBS) at varying serum media concentrations: 5%, 10%, and 15%. Cell viability and density were determined; ELISAs were used to assess SOD1 and TNFα; TaqMan gene array plates were used to explore mRNA gene expression related to growth and atrophy. <b>Results:</b> Cell viability (%) analysis showed that 0% concentration, 12 h incubation, and FBS media have lower viability (<i>p</i> ≤ 0.0001); cell density analysis showed lower values in 0% concentration and in the FBS media (<i>p</i> ≤ 0.0001); SOD1 analysis showed a scaled effect (<i>p</i> ≤ 0.05) and higher concentration in HS (12,795.07 ± 677.87 pg/mL; <i>p</i> ≤ 0.0001); TNFα concentration was higher in HSMM compared to D-HSMM (61 ± 0.82 vs. 2.52 ± 0.94 pg/mL; <i>p</i>=0.017), higher at 12 h (6.07 ± 0.88 vs. 2.50 ± 0.88 pg/mL; <i>p</i>=0.006), and higher in FBS (6.05 ± 0.88 vs. 2.08 ± 0.88 pg/mL; <i>p</i>=0.002); no meaningful increase in fold change was seen in mRNA. <b>Conclusions:</b> Myoblast viability and density were lower in the nutrient-deprived conditions and in the FBS compared to the HS serum. The biomarker of oxidative stress was lower in the serum concentration in a scaled effect, yet higher in HS. The biomarker of inflammation was higher in the HSMM cell line, shorter incubation time period, and in FBS. HS used as a media may supply nutrients and hormonal confounders that may support or stress myoblast growth.</p>","PeriodicalId":16587,"journal":{"name":"Journal of Nutrition and Metabolism","volume":"2025 ","pages":"6661176"},"PeriodicalIF":2.4000,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12187435/pdf/","citationCount":"0","resultStr":"{\"title\":\"The Effect of Nutrient Deprivation on Markers of Oxidative Stress, Inflammation, and Transcriptome in Normal and Type-2 Diabetic Human Skeletal Muscle Myoblasts.\",\"authors\":\"Lael Ceriani, Daniel E Newmire, Xavier F Gonzales, Jean Sparks, Jose Guardiola, Felix O Omoruyi\",\"doi\":\"10.1155/jnme/6661176\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Background:</b> Intermittent fasting has become a new fad diet that may promote an environment to facilitate muscle atrophy, placing aging, and diabetic populations at risk for muscle loss due to nutrient deprivation. The purpose of this study was to investigate how nutrient availability and serum environment influence Type 2 diabetic myoblast density and viability, autophagy-related oxidative and inflammatory markers, and upstream gene expression signaling relevant to proteostasis. <b>Methods:</b> To explore this outcome in human skeletal muscle myoblast (HSMM) and diabetic human skeletal muscle myoblast (D-HSMM), myoblast lines were cultured per standard protocol and were incubated for 12 or 24 h with either human serum (HS) or fetal bovine serum (FBS) at varying serum media concentrations: 5%, 10%, and 15%. Cell viability and density were determined; ELISAs were used to assess SOD1 and TNFα; TaqMan gene array plates were used to explore mRNA gene expression related to growth and atrophy. <b>Results:</b> Cell viability (%) analysis showed that 0% concentration, 12 h incubation, and FBS media have lower viability (<i>p</i> ≤ 0.0001); cell density analysis showed lower values in 0% concentration and in the FBS media (<i>p</i> ≤ 0.0001); SOD1 analysis showed a scaled effect (<i>p</i> ≤ 0.05) and higher concentration in HS (12,795.07 ± 677.87 pg/mL; <i>p</i> ≤ 0.0001); TNFα concentration was higher in HSMM compared to D-HSMM (61 ± 0.82 vs. 2.52 ± 0.94 pg/mL; <i>p</i>=0.017), higher at 12 h (6.07 ± 0.88 vs. 2.50 ± 0.88 pg/mL; <i>p</i>=0.006), and higher in FBS (6.05 ± 0.88 vs. 2.08 ± 0.88 pg/mL; <i>p</i>=0.002); no meaningful increase in fold change was seen in mRNA. <b>Conclusions:</b> Myoblast viability and density were lower in the nutrient-deprived conditions and in the FBS compared to the HS serum. The biomarker of oxidative stress was lower in the serum concentration in a scaled effect, yet higher in HS. The biomarker of inflammation was higher in the HSMM cell line, shorter incubation time period, and in FBS. HS used as a media may supply nutrients and hormonal confounders that may support or stress myoblast growth.</p>\",\"PeriodicalId\":16587,\"journal\":{\"name\":\"Journal of Nutrition and Metabolism\",\"volume\":\"2025 \",\"pages\":\"6661176\"},\"PeriodicalIF\":2.4000,\"publicationDate\":\"2025-06-10\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12187435/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Nutrition and Metabolism\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1155/jnme/6661176\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"2025/1/1 0:00:00\",\"PubModel\":\"eCollection\",\"JCR\":\"Q3\",\"JCRName\":\"NUTRITION & DIETETICS\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Nutrition and Metabolism","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1155/jnme/6661176","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/1/1 0:00:00","PubModel":"eCollection","JCR":"Q3","JCRName":"NUTRITION & DIETETICS","Score":null,"Total":0}
引用次数: 0
摘要
背景:间歇性禁食已经成为一种新的时尚饮食,它可能促进肌肉萎缩的环境,使老年人和糖尿病人群由于营养剥夺而面临肌肉损失的风险。本研究的目的是研究营养供应和血清环境如何影响2型糖尿病成肌细胞密度和活力,自噬相关的氧化和炎症标志物,以及与蛋白质稳态相关的上游基因表达信号。方法:为了在人骨骼肌成肌细胞(HSMM)和糖尿病人骨骼肌成肌细胞(D-HSMM)中探索这一结果,根据标准方案培养成肌细胞系,并在不同的血清培养基浓度(5%,10%和15%)下与人血清(HS)或胎牛血清(FBS)孵育12或24小时。测定细胞活力和密度;elisa法检测SOD1和TNFα;TaqMan基因阵列板检测与生长萎缩相关的mRNA基因表达。结果:细胞活力(%)分析显示,0%浓度、孵育12 h、FBS培养基的细胞活力较低(p≤0.0001);细胞密度分析显示,0%浓度和FBS培养基中细胞密度较低(p≤0.0001);SOD1分析显示比例效应(p≤0.05),HS中SOD1浓度较高(12795.07±677.87 pg/mL);P≤0.0001);HSMM中TNFα浓度高于D-HSMM(61±0.82比2.52±0.94 pg/mL;在12 h p = 0.017),高(6.07±0.88和2.50±0.88 pg / mL;p=0.006), FBS组更高(6.05±0.88比2.08±0.88 pg/mL;p = 0.002);mRNA的折叠变化未见明显增加。结论:与HS血清相比,营养剥夺条件下和FBS中成肌细胞活力和密度较低。氧化应激生物标志物在血清浓度中呈比例效应降低,而在HS中呈比例效应升高。炎症生物标志物在HSMM细胞系中较高,孵育时间较短,FBS中较高。HS作为培养基可以提供营养和激素混杂物,支持或刺激成肌细胞生长。
The Effect of Nutrient Deprivation on Markers of Oxidative Stress, Inflammation, and Transcriptome in Normal and Type-2 Diabetic Human Skeletal Muscle Myoblasts.
Background: Intermittent fasting has become a new fad diet that may promote an environment to facilitate muscle atrophy, placing aging, and diabetic populations at risk for muscle loss due to nutrient deprivation. The purpose of this study was to investigate how nutrient availability and serum environment influence Type 2 diabetic myoblast density and viability, autophagy-related oxidative and inflammatory markers, and upstream gene expression signaling relevant to proteostasis. Methods: To explore this outcome in human skeletal muscle myoblast (HSMM) and diabetic human skeletal muscle myoblast (D-HSMM), myoblast lines were cultured per standard protocol and were incubated for 12 or 24 h with either human serum (HS) or fetal bovine serum (FBS) at varying serum media concentrations: 5%, 10%, and 15%. Cell viability and density were determined; ELISAs were used to assess SOD1 and TNFα; TaqMan gene array plates were used to explore mRNA gene expression related to growth and atrophy. Results: Cell viability (%) analysis showed that 0% concentration, 12 h incubation, and FBS media have lower viability (p ≤ 0.0001); cell density analysis showed lower values in 0% concentration and in the FBS media (p ≤ 0.0001); SOD1 analysis showed a scaled effect (p ≤ 0.05) and higher concentration in HS (12,795.07 ± 677.87 pg/mL; p ≤ 0.0001); TNFα concentration was higher in HSMM compared to D-HSMM (61 ± 0.82 vs. 2.52 ± 0.94 pg/mL; p=0.017), higher at 12 h (6.07 ± 0.88 vs. 2.50 ± 0.88 pg/mL; p=0.006), and higher in FBS (6.05 ± 0.88 vs. 2.08 ± 0.88 pg/mL; p=0.002); no meaningful increase in fold change was seen in mRNA. Conclusions: Myoblast viability and density were lower in the nutrient-deprived conditions and in the FBS compared to the HS serum. The biomarker of oxidative stress was lower in the serum concentration in a scaled effect, yet higher in HS. The biomarker of inflammation was higher in the HSMM cell line, shorter incubation time period, and in FBS. HS used as a media may supply nutrients and hormonal confounders that may support or stress myoblast growth.
期刊介绍:
Journal of Nutrition and Metabolism is a peer-reviewed, Open Access journal that publishes original research articles, review articles, and clinical studies covering the broad and multidisciplinary field of human nutrition and metabolism. The journal welcomes submissions on studies related to obesity, diabetes, metabolic syndrome, molecular and cellular biology of nutrients, foods and dietary supplements, as well as macro- and micronutrients including vitamins and minerals.