Correlation study on circadian rhythm disturbance and pathological characteristics of non-alcoholic fatty liver disease

doi:10.16138/j.1673-6087.2025.03.07

Abstract

Abstract:

Objective To investigate the key signal molecules in the regulation of biological clock and lipid metabolism of non-alcoholic fatty liver disease（NAFLD）and its effect on lipid metabolism, to provide insights for the prevention and treatment of NAFLD. Methods The animal models with NAFLD were established and classified, including circadian rhythm disorder + high-fat diet(HFC) group, circadian rhythm disorder + normal diet(NC) group, high-fat diet (HF) group and normal diet (N) group. Hematoxylin and eosin (HE) and oil red O staining were used to detect the fat deposition in the model liver tissues; enzyme-linked immunosorbent assay(ELISA) was used to detect the serum lipids in the mice; immunoblotting was performed to detect the protein expression of brain and muscle arnt-like 1(BMAL1) gene，and the correlation between BMAL1 gene and the liver pathological features was estimate. For the HFC and HF groups, mRNA bioinformatics analysis was performed to identify key circadian clock genes in NAFLD. Results Circadian rhythm disturbance increased body weight and induced obesity in mice. At week 15, the HFC group showed significantly higher weight than the HF group (t=23.18, P<0.000 1), and the NC group exceeded the N group (t=5.24, P<0.000 1). It also promoted hepatic lipid deposition: lipid content progressively increased in the HFC group (F=10.13, P<0.05) and NC group (F=8.89, P<0.05) over time. Moreover, it exacerbated dyslipidemia: TC and LDL-C levels in the HFC group were significantly higher than the HF group at ZT0, ZT8 and ZT16 (F=23.3, P<0.0001; F=68.1, P<0.000 1); similarly, the NC group had elevated TC and LDL-C versus the N group (F=3.9, P<0.000 1; F=5.8, P<0.000 1). BMAL1 expression exhibited rhythmic fluctuations, with higher protein levels at ZT16 than ZT8 in HFC and NC groups, showing a significant positive correlation with fatty liver severity (r=0.995, P=0.022). Conclusions A high-fat diet causes abnormal lipid metabolism in mice; the disturbance of circadian rhythm exacerbates the abnormal lipid metabolism in mice, increases lipid deposition in the liver, and promotes the progression of fatty liver. The biological clock gene BMAL1 is closely related to the metabolism of non-alcoholic fatty liver disease. High expression of BMAL1 may induce fat accumulating in the liver.

Key words: Non-alcoholic fatty liver disease, Circadian rhythm, BMAL1, Lipid metabolism

CLC Number:

R575.5

HUANG Lei, ZHANG Chenli, YAN Hua, SHI Dongmei. Correlation study on circadian rhythm disturbance and pathological characteristics of non-alcoholic fatty liver disease[J]. Journal of Internal Medicine Concepts & Practice, 2025, 20(03): 224-231.

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References 17

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组别	体重（g）
N组	24.57±2.70
NC组	26.28±2.80
HF组	29.83±5.06
HFC组	32.14±6.69
F	42.59
P	<0.000 1

脂肪沉积率	ZT0	ZT8	ZT16
N组	7.82±6.17	12.45±5.23	10.17±7.15
NC组	8.67±6.76	16.24±6.47	33.14±8.45
HF组	34.5±7.86	32.86±5.46	30.17±9.17
HFC组	10.62±6.7	21.45±9.74	38.72±6.22
F	7.87
P	<0.000 1

血脂		ZT0	ZT8	ZT16	F	P
TC					66.3	<0.000 1
	N组	232.5±14.4	212.7±9.7	220.7±4.7
	NC组	263.3±7.5	251.0±27.0	261.1±31.8
	HF组	351.0±9.9^1）	355.8±8.4^1）	332.0±17.7^1）
	HFC组	447.8±26.3^2）	428.1±22.0^2）	445.6±22.2^2）
TG					79.5	<0.000 1
	N组	270.5±23.9	273.8±10.9	276.0±10.3
	NC组	278.3±25.3	244.6±11.6	261.3±7.6
	HF组	420.4±18.5^1）	474.0±6.3^1）	339.8±13.7^1）
	HFC组	449.5±33.2^2）	487.5±13.3^2）	330.1±15.7^2）
LDL-C					2313.0	<0.000 1
	N组	224.3±11.5	227.9±15.8	240.8±23.5
	NC组	256.1±29.0	273.5±10.8	287.7±10.5
	HF组	6 239.7±64.4^1）	6 058.6±81.7^1）	71 26.4±302.2^1）
	HFC组	7 193.4±211.1^2）	6 635.0±122.4^2）	8 422.1±177.9^2）
HDL-C					454.0	<0.000 1
	N组	218.3±11.5	226.7±7.0	229.6±11.2
	NC组	201.7±19.7	232.5±18.4	219.0±16.9
	HF组	2 461.0±156.2^1）	2 484.7±82.5^1）	2 289.1±79.2^1）
	HFC组	2 528.0±204.3^2）	2 492.9±96.4^2）	2 417.7±131.9^2）

组别	ZT0	ZT8	ZT16
N组	0.7±0.1	0.3±0.0	0.3±0.0
HF组	0.9±0.0^1）	0.6±0.0^1）	0.5±0.0^2）
F	175.6
P	<0.000 1
NC组	0.8±0.0	0.4±0.0	0.6±0.0
HFC组	0.9±0.0^3）	0.5±0.0^4）	0.8±0.0^4）
F	611.9
P	<0.000 1

基因	ZT0			ZT8			ZT16
基因	HF组		HFC组	HF组	HFC组		HF组		HFC组
BMAL1	592	862		487		364	352	953
BMAL2	10	25		38		32	28	29
CLOCK	2 007	2 918		2 526		2 327	2 233	2 468
PER1	418	750		153		409	237	515
PER2	848	834		345		949	545	843
PER3	328	214		297		1 109	578	251
NPAS2	44	147		144		65	73	305
CRY1	843	1 053		289		374	294	1 174
CRY2	698	795		733		1 199	894	871