目的 验证不同时间补钙是否会对心肌产生影响,为科学补钙提供依据。方法 60只SPF级雌性ICR小鼠分为早上对照组（MC）、晚上对照组（EC）、早上补钙组（MS）、晚上补钙组（ES）。补钙组喂饲钙含量0.15%的饲料,并经口灌胃6.75 g/（kg·d）的碳酸钙溶液;对照组喂饲钙含量1.5%的饲料,并经口灌胃等体积的水。10 w后处死小鼠,测定血清肿瘤坏死因子α(tumor necrosis factor , TNF-α)、白细胞介素6(interleukin 6, IL-6)、心肌损伤指标心肌肌钙蛋白（cardiac troponin, cTNT）、超氧化物歧化酶（superoxide dismutase, SOD）、丙二醛（malondialdehyde, MDA）和血钙动力学,计算心体比,染色观察心脏病理,转录组测序、实时荧光定量PCR（RT-qPCR）和蛋白免疫印迹（Western blot）测定心肌相关基因和蛋白表达变化。结果 与ES组相比,MS组心体比指数升高（P<0.01）,MS和ES组血清MDA含量升高（P<0.05）,且MS组MDA升高更显著,MS组SOD活力呈下降趋势。MS组血清TNF-α、cTNT、IL-6明显升高（P<0.05）。MS组心肌细胞排列明显紊乱,细胞核肥大,有炎症细胞浸润、坏死现象。早、晚补钙组差异表达基因多富集在PI3K-Akt信号通路、MAPK信号通路、钙信号通路等通路。PCR与蛋白表达分析结果显示,与ES组比MS组Gadd45b、Plcb4表达上升,Cacna1g表达下降。结论 早上补钙会激活氧化应激反应,加剧炎症作用,促使心肌肥厚、心脏受损。
Abstract
Objective To investigate whether calcium supplementation at different times of the day affects myocardial health, providing a scientific basis for calcium supplementation. Methods Sixty SPF-grade female ICR mice were divided into four groups: morning control (MC), evening control (EC), morning calcium supplementation (MS), and evening calcium supplementation (ES). The calcium-supplemented groups were fed a 0.15% calcium diet and orally administered 6.75 g/(kg·d) of calcium carbonate solution, while the control groups were fed a 1.5% calcium diet and given an equal volume of water. After 10 weeks, the mice were euthanized, and serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), cardiac troponin (cTNT), superoxide dismutase (SOD), malondialdehyde (MDA), and calcium kinetics were measured. The heart-to-body weight ratio was calculated, and cardiac pathology was assessed via histopathological staining. Transcriptome sequencing, real-time quantitative PCR (RT-qPCR), and Western blot were performed to analyze changes in myocardial-related gene and protein expression. Results Compared with the ES group, the MS group exhibited a significantly increased heart-to-body weight ratio (P<0.01). Both MS and ES groups showed elevated serum MDA levels (P<0.05), with a more pronounced increase in the MS group, while SOD activity tended to decrease in the MS group. Serum TNF-α, cTNT, and IL-6 levels were significantly higher in the MS group (P<0.05). Histopathological analysis revealed disordered myocardial cell arrangement, hypertrophic nuclei, inflammatory cell infiltration, and necrosis in the MS group. Differentially expressed genes between morning and evening calcium-supplemented groups were primarily enriched in the PI3K-Akt, MAPK, and calcium signaling pathways. RT-qPCR and Western blot analyses demonstrated that, compared with the ES group, the MS group exhibited upregulated expression of Gadd45b and Plcb4 and downregulated expression of Cacna1g. Conclusion Morning calcium supplementation activates oxidative stress, exacerbates inflammation, and promotes myocardial hypertrophy and cardiac injury.
关键词
钙 /
昼夜节律 /
心脏损伤 /
时序营养 /
转录组学
Key words
calcium /
circadian rhythm /
cardiac damage /
chrononutrition /
transcriptomics
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基金
黑龙江省自然科学基金（No.LH2022H050）
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