Differences of calorie restriction and time-restricted feeding on metabolic indices and gut microbiota of mice

doi:10.12307/2025.907

Abstract

Abstract: BACKGROUND: Both calorie restriction and time-restricted feeding, as two common dietary patterns, have been shown to improve health by regulating metabolism. However, the difference between these dietary patterns, metabolic indices, as well as the gut microbiota still requires further attention.

OBJECTIVE: To explore the differences of calorie restriction and time-restricted feeding on the metabolic indices and gut microbiota of mice.
METHODS: The C57BL/6J mice were randomly divided into three groups of ad libitum, calorie restriction, and time-restricted feeding (n=6 per group) for 28 weeks of dietary intervention. Various parameters such as body weight, food intake, glucose tolerance, serum fasting insulin, Homeostasis Model Assessment of Insulin Resistance, and leptin were measured. The impact of different interventions on the gut microbiota structure in mice was explored using 16S rRNA sequence analysis. Key operational taxonomic units responsive to dietary interventions were identified through LEfSe analysis.
RESULTS AND CONCLUSION: (1) Compared with the ad libitum group, the body weight, food intake, area under the glucose tolerance curve of the calorie restriction and time-restricted feeding groups were decreased (P < 0.01), and the serum leptin was decreased (P < 0.05). The fasting insulin level and serum leptin level of the calorie restriction group were decreased (P < 0.05) and were significantly lower than those of the time-restricted feeding group (P < 0.05); homeostasis model assessment of insulin resistance decreased in the calorie restriction group (P < 0.01). (2) Compared with the ad libitum group, the α diversity of gut microbiota in the calorie restriction group and the time-restricted feeding group was decreased (P < 0.05), but the diversity of the time-restricted feeding group was slightly lower than that in the calorie restriction group. (3) There were 15 key operational taxonomic units related to calorie restriction and the time-restricted feeding intervention, of which 8 were positively correlated with metabolic phenotypes and their abundance decreased, and 3 were negatively correlated with metabolic phenotypes and their abundance increased (P < 0.05). OTU819 Lachnospiraceae_UCG-006 was positively correlated with body weight, area under the glucose tolerance curve, homeostasis model assessment of insulin resistance, and fasting insulin, while OTU1397 Muribaculaceae was negatively correlated with these indicators. The results show that both calorie restriction and the time-restricted feeding intervention can improve the weight and glucose metabolism of mice, and both intervention modes caused the remodeling of the gut microbiota, which helped to improve the metabolic disorders.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: calorie restriction, time-restricted feeding, glucose metabolism, fasting insulin, metabolic phenotype, gut microbiota, operational taxonomic units, Ileibacterium_valens

CLC Number:

Cui Yuena, Chen Xiaoyu, Liang Meiting, Chen Wujin, He Yi, Dilinur·Ekpa, Du Manxi, Zhu Yuqiu, Abuduwupuer·Haibier, Sun Yuping. Differences of calorie restriction and time-restricted feeding on metabolic indices and gut microbiota of mice[J]. Chinese Journal of Tissue Engineering Research, 2025, 29(30): 6449-6456.

Figures/Tables 9

2.3.2 β多样性比较限时进食组小鼠的肠道菌群趋向于节食组，与自由取食组在PC1轴上显著分开，该方向能解释整体菌群变异度的31.46%；而节食组小鼠的肠道菌群则在PC2轴上与自由取食组小鼠的肠道菌群存在显著差异，该方向能解释整体菌群变异度的21.61%，见图2，提示节食和限时进食小鼠的肠道菌群整体结构发生改变。 2.3.3 门水平组成比较对丰度在1%以上的7个优势门进行分析，发现与自由取食组相比，节食组放线菌的相对丰度增加，弯曲菌门的相对丰度减少；限时进食组厚壁菌门的相对丰度增加，拟杆菌、弯曲菌门的相对丰度减少；与节食组相比，限时进食组厚壁菌门、髌骨细菌门、脱硫杆菌门的相对丰度增加，差异均有显著性意义(P < 0.05)，见图3。 2.3.4 属水平组成比较对丰度在1%以上的21个优势属进行分析，发现与自由取食组相比，节食组回肠杆菌属、双歧杆菌属丰度升高，毛螺菌科_NK4A136、梭状芽胞杆菌目_UCG-014、毛螺菌科的未知属、颤螺菌科的未知属、另枝菌属、螺杆菌属、葡聚糖菌属、苏黎世杆菌、罗斯氏菌丰度降低；限时进食组乳酸菌属丰度升高，Muribaculaceae科的未知属、梭状芽胞杆菌目_UCG-014、颤螺菌科的未知属、另枝菌属、毛螺菌科的未知属、葡聚糖菌属、苏黎世杆菌属、罗斯氏菌属丰度降低。与节食组相比，限时进食组乳酸菌属、萨科瑞莫纳斯属、脱硫弧菌属丰度显著升高，回肠杆菌属、双歧杆菌属丰度降低，以上差异均有显著性意义(P < 0.05)，见图4。 2.3.5 肠道菌群LEfSe分析找出关键操作分类单元通过LEfSe分析在操作分类单元水平上发现15个可能是节食和限时进食干预的关键操作分类单元，这些操作分类单元分别属于Muribaculaceae(5个操作分类单元)、毛螺菌科(3个操作分类单元，1个属于Lachnoclostridium，1个属于毛螺菌科_UCG-006)、乳杆菌属(2个操作分类单元，其中1个属于鼠乳杆菌，1个属于肠乳杆菌)、颤螺菌科(2个操作分类单元，其中1个属于葡聚糖菌属)、Helicobacter_ganmani(1个操作分类单元)、红蝽菌科_UCG-002(1个操作分类单元)、Ileibacterium_valens(1个操作分类单元)。LDA得分最高的为OTU836鼠乳杆菌，数值为5.22，其次是OTU810 Ileibacterium_valens，数值为4.95，见图5。 2.3.6 关键操作分类单元与代谢表型关联分析将15个关键操作分类单元与小鼠代谢表型进行Spearman相关分析，发现11个操作分类单元与小鼠代谢表型相关，其中8个呈正相关，3个呈负相关(P < 0.05)。呈正相关的操作分类单元中OTU735 Helicobacter_ganmani，OTU762葡聚糖菌属，OTU819 毛螺菌科_UCG-006及Muribaculaceae科的OTU1330、OTU3848和OTU3897与体质量和腹腔注射葡萄糖耐量实验曲线下面积呈正相关，Muribaculaceae科的OTU1428和OTU3848与空腹血糖呈正相关OTU1333 Lachnoclostridium仅与体质量呈正相关，OTU810 Ileibacterium_valens仅与体质量呈负相关，OTU836鼠乳杆菌与空腹血糖呈负相关；11个指标中最值得关注的是OTU819毛螺菌科_UCG-006，与体质量、腹腔注射葡萄糖耐量实验曲线下面积、HOMA-IR、空腹胰岛素均呈正相关，而OTU1397 Muribaculaceae与以上指标均呈负相关，见图6。 2.3.7 关键操作分类单元丰度变化在与代谢表型呈正相关的8个操作分类单元中，节食组和限时进食组中5个操作分类单元(Muribaculaceae科的OTU1330和OTU3897、OTU735 Helicobacter_ganmani，OTU762葡聚糖菌属、OTU1333 Lachnoclostridium)丰度占比均低于自由取食组，OTU819毛螺菌科_UCG-006的丰度占比仅在节食组显著降低，Muribaculaceae科的OTU1428和OTU3848丰度占比仅在限时进食组显著降低。与节食组相比，OTU1333 Lachnoclostridium的丰度占比在限时进食组中升高，OTU1428 Muribaculaceae的丰度占比在限时进食组中降低。在与代谢表型呈负相关的3个操作分类单元中，OTU836鼠乳杆菌在节食组和限时进食组丰度占比均显著升高，分别高达17.23% (8.00%，31.01%)，37.87% (24.06%，53.42%)，OTU810 Ileibacterium_valens仅在节食组中显著升高，丰度占比22.14%(12.53%，24.60%)，OTU1397 Muribaculaceae仅在节食组中显著升高。与节食组相比，OTU810 Ileibacterium_valens、OTU1397 Muribaculaceae的丰度占比在限时进食组中降低，以上差异均有显著性意义(P < 0.05)，见表4。"

References

[1] LONGO VD, ANDERSON RM. Nutrition, longevity and disease: From molecular mechanisms to interventions. Cell. 2022;185(9):1455-1470.
[2] DI FRANCESCO A, DI GERMANIO C, BERNIER M, et al. A time to fast. Science. 2018;362(6416):770-775.
[3] YAMASHITA S, IMAI S, MOMO K, et al. Investigation of the real-world situation and risk factors associated with olanzapine prescribed to diabetes patients by using a japanese claims database. Biol Pharm Bull. 2021;44(8):1151-1155.
[4] DUREGON E, POMATTO-WATSON LCDD, BERNIER M, et al. Intermittent fasting: from calories to time restriction. Geroscience. 2021;43(3): 1083-1092.
[5] KORD-VARKANEH H, SALEHI-SAHLABADI A, TINSLEY GM, et al. Effects of time-restricted feeding (16/8) combined with a low-sugar diet on the management of non-alcoholic fatty liver disease: A randomized controlled trial. Nutrition. 2023;105:111847.
[6] 梁磊,念馨.肠道菌群:未来2型糖尿病治疗的新靶点[J].实用医学杂志,2022,38(3):261-265.
[7] 张静,王肖枭,周怡,等.肠道菌群与疾病相关性的研究进展[J].基础医学与临床,2020,40(2):243-247.
[8] ADAK A, KHAN MR. An insight into gut microbiota and its functionalities. Cell Mol Life Sci. 2019;76(3):473-493.
[9] WU J, YANG L, LI S, et al. Metabolomics Insights into the Modulatory Effects of Long-Term Low Calorie Intake in Mice. J Proteome Res. 2016;15(7):2299-2308.
[10] ZHANG L, XUE X, ZHAI R, et al. Timing of Calorie Restriction in Mice Impacts Host Metabolic Phenotype with Correlative Changes in Gut Microbiota. mSystems. 2019;4(6):e00348-19.
[11] PIECZYŃSKA-ZAJĄC JM, MALINOWSKA A, ŁAGOWSKA K, et al. The effects of time-restricted eating and Ramadan fasting on gut microbiota composition: a systematic review of human and animal studies. Nutr Rev. 2024;82(6):777-793.
[12] 潘凤伟.节食小鼠肠道中优势乳杆菌的分离鉴定及其在衰老相关炎症中的作用[D].上海:上海交通大学,2018.
[13] SUN J, SHE Y, FANG P, et al. Time-restricted feeding prevents metabolic diseases through the regulation of galanin/GALR1 expression in the hypothalamus of mice. Eat Weight Disord. 2022;27(4):1415-1425.
[14] TSITSOU S, ZACHARODIMOS N, POULIA KA, et al. Effects of Time-Restricted Feeding and Ramadan Fasting on Body Weight, Body Composition, Glucose Responses, and Insulin Resistance: A Systematic Review of Randomized Controlled Trials. Nutrients. 2022;14(22):4778.
[15] MAIFELD A, BARTOLOMAEUS H, LÖBER U, et al. Fasting alters the gut microbiome reducing blood pressure and body weight in metabolic syndrome patients. Nat Commun. 2021;12(1):1970.
[16] LI L, SU Y, LI F, et al. The effects of daily fasting hours on shaping gut microbiota in mice. BMC Microbiol. 2020;20(1):65.
[17] GOH KL, CHAN WK, SHIOTA S, et al. Epidemiology of Helicobacter pylori infection and public health implications. Helicobacter. 2011;16 Suppl 1(0 1):1-9.
[18] LI X, LV H, SHI F, et al. The potential therapeutic effects of hydroxypropyl cellulose on acute murine colitis induced by DSS. Carbohydr Polym. 2022;289:119430.
[19] GUO J, WANG P, CUI Y, et al. Protective Effects of Hydroxyphenyl Propionic Acids on Lipid Metabolism and Gut Microbiota in Mice Fed a High-Fat Diet. Nutrients. 2023;15(4):1043.
[20] GUO M, XING D, WANG J, et al. Potent Intestinal Mucosal Barrier Enhancement of Nostoc commune Vaucher Polysaccharide Supplementation Ameliorates Acute Ulcerative Colitis in Mice Mediated by Gut Microbiota. Nutrients. 2023;15(13):3054.
[21] CHEN H, ZHAO H, QI X, et al. Lactobacillus plantarum HF02 alleviates lipid accumulation and intestinal microbiota dysbiosis in high-fat diet-induced obese mice. J Sci Food Agric. 2023;103(9):4625-4637.
[22] ZHANG F, ZHOU Y, CHEN H, et al. Curcumin Alleviates DSS-Induced Anxiety-Like Behaviors via the Microbial-Brain-Gut Axis. Oxid Med Cell Longev. 2022;2022:6244757.
[23] HAO H, ZHANG X, TONG L, et al. Effect of Extracellular Vesicles Derived From Lactobacillus plantarum Q7 on Gut Microbiota and Ulcerative Colitis in Mice. Front Immunol. 2021;12:777147.
[24] WANG J, HAN L, LIU Z, et al. Genus unclassified_Muribaculaceae and microbiota-derived butyrate and indole-3-propionic acid are involved in benzene-induced hematopoietic injury in mice. Chemosphere. 2023;313:137499.
[25] SHAO J, LI Z, GAO Y, et al. Construction of a “Bacteria-Metabolites” Co-Expression Network to Clarify the Anti-Ulcerative Colitis Effect of Flavonoids of Sophora flavescens Aiton by Regulating the “Host-Microbe” Interaction. Front Pharmacol. 2021;12:710052.
[26] LIU Y, ZHOU M, YANG M, et al. Pulsatilla chinensis Saponins Ameliorate Inflammation and DSS-Induced Ulcerative Colitis in Rats by Regulating the Composition and Diversity of Intestinal Flora. Front Cell Infect Microbiol. 2021;11:728929.
[27] MO J, NI J, ZHANG M, et al. Mulberry Anthocyanins Ameliorate DSS-Induced Ulcerative Colitis by Improving Intestinal Barrier Function and Modulating Gut Microbiota. Antioxidants (Basel). 2022;11(9):1674.
[28] YUAN G, TAN M, CHEN X. Punicic acid ameliorates obesity and liver steatosis by regulating gut microbiota composition in mice. Food Funct. 2021;12(17):7897-7908.
[29] ZHAO D, CAO J, JIN H, et al. Beneficial impacts of fermented celery (Apium graveolens L.) juice on obesity prevention and gut microbiota modulation in high-fat diet fed mice. Food Funct. 2021;12(19):9151-9164.
[30] SONG H, SHEN X, CHU Q, et al. Vaccinium bracteatum Thunb. fruit extract reduces high-fat diet-induced obesity with modulation of the gut microbiota in obese mice. J Food Biochem. 2021;45(7):e13808.
[31] SLATTERY C, COTTER PD, O’TOOLE PW. Analysis of Health Benefits Conferred by Lactobacillus Species from Kefir. Nutrients. 2019;11(6):1252.
[32] DEMPSEY E, CORR SC. Lactobacillus spp. for Gastrointestinal Health: Current and Future Perspectives. Front Immunol. 2022;13:840245.
[33] 陈晓瑜,陈峰,张晨虹.一株节食小鼠肠道优势鼠乳杆菌加速DSS致慢性结肠炎恢复[J].基因组学与应用生物学,2021,40(2):855-866.
[34] RANGAN P, CHOI I, WEI M, et al. Fasting-Mimicking Diet Modulates Microbiota and Promotes Intestinal Regeneration to Reduce Inflammatory Bowel Disease Pathology. Cell Rep. 2019;26(10):2704-2719.e6.
[35] QIU J, CHEN L, ZHANG L, et al. Xie Zhuo Tiao Zhi formula modulates intestinal microbiota and liver purine metabolism to suppress hepatic steatosis and pyroptosis in NAFLD therapy. Phytomedicine. 2023; 121:155111.
[36] LU X, QI C, ZHENG J, et al. The Antidepressant Effect of Deoiled Sunflower Seeds on Chronic Unpredictable Mild Stress in Mice Through Regulation of Microbiota-Gut-Brain Axis. Front Nutr. 2022;9:908297.
[37] JI S, HAN S, YU L, et al. Jia Wei Xiao Yao San ameliorates chronic stress-induced depression-like behaviors in mice by regulating the gut microbiome and brain metabolome in relation to purine metabolism. Phytomedicine. 2022;98:153940.