异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响

doi:10.3969/j.issn.1006-9771.2022.05.010

《中国康复理论与实践》 ›› 2022, Vol. 28 ›› Issue (5): 552-558.doi: 10.3969/j.issn.1006-9771.2022.05.010

异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响

宋绍霏¹,侯园园¹,王云雷¹,张通^1,²()

1.首都医科大学康复医学院,北京市 100068
2.中国康复研究中心北京博爱医院神经康复科,北京市 100068

收稿日期:2022-01-05 修回日期:2022-04-27 出版日期:2022-05-25 发布日期:2022-06-10
通讯作者: 张通 E-mail:tom611@126.com
作者简介:宋绍霏(1997-),女,汉族,四川乐山市人,硕士研究生,主要研究方向：昼夜节律紊乱、神经康复
基金资助:
1. 中国康复研究中心基金课题(2020-07);2. 中央级公益性科研院所基本科研业务费专项资金项目(2019CZ-2)

Effects of circadian misalignment induced by abnormal photoperiod on expression rhythm of clock genes and glucose uptake related genes in gastrocnemius in rats

SONG Shaofei¹,HOU Yuanyuan¹,WANG Yunlei¹,ZHANG Tong^1,²()

1. Capital Medical University School of Rehabilitation Medicine, Beijing 100068, China
2. Department of Neurological Rehabilitation, Beijing Bo'ai Hospital, China Rehabilitation Research Center, Beijing 100068, China

Received:2022-01-05 Revised:2022-04-27 Published:2022-05-25 Online:2022-06-10
Contact: ZHANG Tong E-mail:tom611@126.com
Supported by:
China Rehabilitation Research Center Research Project(2020-07);Fundamental Research Funds for Central Public Welfare Research Institutes(2019CZ-2)

摘要/Abstract

摘要：

目的探讨昼夜节律紊乱(CM)对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响。方法雄性Wistar大鼠36只分为昼夜节律正常(CA)组和CM组,每组18只。造模第61天行ClockLab行为学检测。造模第85天行腹腔注射糖耐量试验和生理指标检测。第91~92天于8:00、12:00、16:00、20:00、24:00、4:00处死大鼠,收集腓肠肌组织,逆转录实时定量聚合酶链反应(RT-qPCR)检测Bmal1、Clock、Per2、Tbc1d1、Glut4、Pgc1α的mRNA。结果 CM组昼夜节律周期延长(t = -6.557, P < 0.001)、振幅下降(t = 2.326, P = 0.030),腹腔注射糖耐量试验血糖曲线下面积减少(t = -2.622, P = 0.016)。腓肠肌不同时间Bmal1 (F = 6.691, P < 0.001)、Clock(F = 4.188, P = 0.007)、Per2 (F = 10.893, P < 0.001)、Tbc1d1 (F = 3.411, P = 0.018)、Glut4 (F = 5.439, P = 0.002)、Pgc1α (F = 15.376, P < 0.001)表达有显著性差异,两组间腓肠肌Bmal1 (F = 5.020, P = 0.035)、Per2(F = 8.996, P = 0.006)、Tbc1d1 (F = 51.111, P < 0.001)、Pgc1α (F = 10.177, P = 0.004)表达有显著性差异,CM组腓肠肌24 h总Tbc1d1表达显著下降(t = 4.349, P < 0.001)。结论异常光周期所致CM可导致大鼠糖耐量下降,这可能与大鼠腓肠肌Tbc1d1表达下降以及Bmal1、Per2、Pgc1α的转录节律改变有关。

关键词: 昼夜节律紊乱, 骨骼肌, 糖耐量, 生物钟, 大鼠

Abstract:

Objective To investigate the impacts of circadian misalignment on glucose uptake of skeletal muscle and metabolism in rats. Methods A total of 36 male Wistar rats were divided into circadian alignment (CA, normal light-dark cycles, n = 18) and circadian misalignment (CM, shifted light-dark cycles, n = 18) groups. ClockLab behavior analysis was performed for 18 days (61 to 78 days after modeling). Intraperitoneal injection glucose tolerance test and physiologic measures were performed 85 days after modeling. They were euthanized 91 to 92 days after modeling, at 8:00, 12:00, 16:00, 20:00, 24:00 and 4:00 next day. Gastrocnemius tissue was collected and measured for Bmal1, Clock, Per2, Tbc1d1, Glut4 and Pgc1α by reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR). Results The circadian cycle increased (t = -6.557, P < 0.001), the amplitude decreased (t = 2.326, P = 0.030) and the area under curve (AUC) of the blood glucose of intraperitoneal glucose tolerance test decreased (t = -2.622, P = 0.016) in CM group. In gastrocnemius, there was difference in the expression of the Bmal1 (F = 6.691, P < 0.001), Clock (F = 4.188, P = 0.007), Per2 (F = 10.893, P < 0.001), Tbc1d1 (F = 3.411, P = 0.018), Glut4 (F = 5.439, P = 0.002) and Pgc1α (F = 15.376, P < 0.001) across different time; meanwhile, there was difference in the expression of Bmal1 (F = 5.020, P = 0.035), Per2 (F = 8.996, P = 0.006), Tbc1d1 (F = 51.111, P < 0.001) and Pgc1α (F = 10.177, P = 0.004) between groups, and the total Tbc1d1 expression decreased in CM group (t = 4.349, P < 0.001). Conclusion CM induced by shifted light-dark cycles may lead to glucose tolerance impairment in rats, which may be related to the decreased expression of Tbc1d1 and the changes of transcription rhythm of Bmal1, Per2 and Pgc1α in gastrocnemius.

Key words: circadian misalignment, skeletal muscle, glucose tolerance, circadian clock, rats

中图分类号:

R587.1

宋绍霏,侯园园,王云雷,张通. 异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响[J]. 《中国康复理论与实践》, 2022, 28(5): 552-558.

SONG Shaofei,HOU Yuanyuan,WANG Yunlei,ZHANG Tong. Effects of circadian misalignment induced by abnormal photoperiod on expression rhythm of clock genes and glucose uptake related genes in gastrocnemius in rats[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2022, 28(5): 552-558.

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组别	n	节律周期/h	节律振幅
CA组	12	24.0±0.1	500.6±120.8
CM组	12	27.0±1.6	412.7±50.2
t值		-6.557	2.326
P值		< 0.001	0.030

组别	n	血糖/mmol·L^-1						AUC
组别	n	0 min	15 min	30 min	60 min	90 min	120 min	AUC
CA组	12	4.7±0.8	14.9±3.3	14.5±4.1	10.4±3.9	8.0±3.5	6.5±2.0	1232.3±362.6
CM组	12	5.2±0.7	19.1±5.1	20.1±6.8	15.0±5.6	10.9±3.2	7.6±1.8	1669.6±449.9
t值		-1.790	-2.445	-2.456	-2.318	-2.159	-1.464	-2.622
P值		0.087	0.024	0.022	0.030	0.042	0.157	0.016

组别	n	体质量/g	腹围/cm	体长/cm	Lee指数
CA组	12	458.1±29.5	19.1±1.0	24.8±3.2	26.5±1.1
CM组	12	468.7±28.9	19.1±1.1	24.9±1.0	26.6±0.5
t值		-0.888	-0.061	-0.086	-0.217
P值		0.384	0.952	0.932	0.831

组别	n	4:00	8:00	12:00	16:00	20:00	24:00	24 h总表达
CA组	3	0.52±0.09	0.89±0.14	0.37±0.16	0.07±0.04	0.11±0.02	0.30±0.11	0.38±0.30
CM组	3	0.10±0.01	0.29±0.09	0.14±0.08	0.52±0.27	0.31±0.18	0.33±0.05	0.28±0.18
t值		7.806	6.351	2.158	-2.881	-2.020	-0.330	1.138
P值		0.001	0.003	0.097	0.045	0.114	0.758	0.263

组别	n	4:00	8:00	12:00	16:00	20:00	24:00	24 h总表达
CA组	3	0.65±0.11	0.79±0.12	1.40±0.41	0.80±0.15	0.68±0.17	0.66±0.16	0.83±0.32
CM组	3	0.47±0.18	0.69±0.08	0.71±0.14	0.74±0.02	0.98±0.23	0.83±0.25	0.74±0.21
t值		1.489	1.188	2.723	0.648	-1.801	-1.000	1.013
P值		0.211	0.301	0.053	0.581	0.146	0.374	0.318

异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响

Effects of circadian misalignment induced by abnormal photoperiod on expression rhythm of clock genes and glucose uptake related genes in gastrocnemius in rats

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组别	n	4:00	8:00	12:00	16:00	20:00	24:00	24 h总表达
CA组	3	1.26±0.43	0.41±0.04	1.04±0.04	4.19±0.71	5.19±1.37	2.45±0.47	2.42±1.88
CM组	3	1.00±0.63	1.73±0.73	3.14±0.87	1.27±0.31	1.57±0.80	1.83±0.35	1.76±0.89
t值		0.603	-3.120	-4.171	6.485	3.954	1.804	1.364
P值		0.579	0.088	0.053	0.003	0.017	0.146	0.185

组别	n	4:00	8:00	12:00	16:00	20:00	24:00	24 h总表达
CA组	3	1.16±0.15	0.80±0.22	0.73±0.09	1.12±0.42	2.11±0.76	1.92±0.12	1.31±0.63
CM组	3	0.30±0.09	0.76±0.16	0.87±0.23	0.84±0.20	0.42±0.19	0.42±0.18	0.60±0.28
t值		8.391	0.217	-0.990	1.062	3.716	11.847	4.349
P值		0.001	0.839	0.378	0.348	0.054	< 0.001	< 0.001

组别	n	4:00	8:00	12:00	16:00	20:00	24:00	24 h总表达
CA组	3	1.13±0.02	0.60±0.12	0.86±0.12	0.98±0.27	1.22±0.19	2.74±0.37	1.26±0.74
CM组	3	1.01±0.16	0.92±0.14	1.11±0.20	0.86±0.25	0.77±0.06	1.18±0.65	0.97±0.30
t值		1.386	-2.968	-1.778	0.591	3.955	3.607	1.517
P值		0.295	0.041	0.150	0.586	0.017	0.023	0.143

组别	n	4:00	8:00	12:00	16:00	20:00	24:00	24 h总表达
CA组	3	0.82±0.12	1.14±0.26	1.06±0.05	0.92±0.14	1.28±0.25	1.49±0.20	1.12±0.28
CM组	3	1.22±0.19	1.54±0.13	1.62±0.11	1.09±0.17	0.91±0.06	0.73±0.04	1.18±0.35
t值		-3.112	-2.397	-8.270	-1.317	2.550	6.608	-0.635
P值		0.036	0.075	0.001	0.258	0.063	0.018	0.530

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