Ceriani Lael, Newmire Daniel E, Gonzales Xavier F, Sparks Jean, Guardiola Jose, Omoruyi Felix O
Department of Life Sciences, Texas A&M University-Corpus Christi, 6300 Ocean Drive, Corpus Christi 78412, Texas, USA.
School of Health Promotion and Kinesiology, Institute for Women's Health, Texas Woman's University, 1600 N Bell Ave, Pioneer Hall, Denton 76209, Texas, USA.
J Nutr Metab. 2025 Jun 10;2025:6661176. doi: 10.1155/jnme/6661176. eCollection 2025.
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. 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. Cell viability (%) analysis showed that 0% concentration, 12 h incubation, and FBS media have lower viability ( ≤ 0.0001); cell density analysis showed lower values in 0% concentration and in the FBS media ( ≤ 0.0001); SOD1 analysis showed a scaled effect ( ≤ 0.05) and higher concentration in HS (12,795.07 ± 677.87 pg/mL; ≤ 0.0001); TNFα concentration was higher in HSMM compared to D-HSMM (61 ± 0.82 vs. 2.52 ± 0.94 pg/mL; =0.017), higher at 12 h (6.07 ± 0.88 vs. 2.50 ± 0.88 pg/mL; =0.006), and higher in FBS (6.05 ± 0.88 vs. 2.08 ± 0.88 pg/mL; =0.002); no meaningful increase in fold change was seen in mRNA. 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.
间歇性禁食已成为一种新的时尚饮食方式，它可能营造出一种促进肌肉萎缩的环境，使老年人和糖尿病患者群体因营养缺乏而面临肌肉流失的风险。本研究的目的是调查营养物质供应和血清环境如何影响2型糖尿病成肌细胞的密度和活力、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。为了在人骨骼肌成肌细胞（HSMM）和糖尿病患者骨骼肌成肌细胞（D-HSMM）中探究这一结果，按照标准方案培养成肌细胞系，并在不同血清培养基浓度（5%、10%和15%）下，用人血清（HS）或胎牛血清（FBS）孵育12或24小时。测定细胞活力和密度；使用酶联免疫吸附测定法（ELISA）评估超氧化物歧化酶1（SOD1）和肿瘤坏死因子α（TNFα）；使用TaqMan基因阵列板探究与生长和萎缩相关的mRNA基因表达。细胞活力（%）分析表明，0%浓度、12小时孵育和FBS培养基的活力较低（≤0.0001）；细胞密度分析表明，0%浓度和FBS培养基中的值较低（≤0.0001）；SOD1分析显示出一种比例效应（≤0.05），且在HS中的浓度较高（12,795.07±677.87 pg/mL；≤0.0001）；与D-HSMM相比，HSMM中的TNFα浓度更高（61±0.82对2.52±0.94 pg/mL；P=0.017），在12小时时更高（6.07±0.88对2.50±0.88 pg/mL；P=0.006），在FBS中更高（6.05±0.88对2.08±0.88 pg/mL；P=0.002）；mRNA的倍数变化未见有意义的增加。与HS血清相比，在营养缺乏条件下和FBS中，成肌细胞的活力和密度较低。氧化应激生物标志物在血清浓度中呈比例效应降低，但在HS中较高。炎症生物标志物在HSMM细胞系、较短孵育时间段和FBS中较高。用作培养基的HS可能提供支持或影响成肌细胞生长的营养物质和激素混杂因素。
