The in vivo and in vitro effects of Eriodictyol on metabolic dysfunction-associated steatotic liver disease by regulating UBA52 expression

doi:10.11958/20252251

Abstract

Abstract:

Objective To investigate the effect of Eriodictyol (ERI) on the development of metabolic dysfunction-associated steatotic liver disease by regulating the expression of ubiquitin A 52 (UBA52) at both in vivo and in vitro levels. Methods A mouse metabolic dysfunction-associated steatotic liver disease model was established using a high-fat diet induction. The mice were randomly separated into the normal control group (normal group), the model group, the low-dose ERI group (ERI-L group, 50 mg/kg ERI) and the high-dose ERI group (ERI-H group, 100 mg/kg ERI), with 12 mice in each group. Oil red O staining was applied to observe lipid deposition in mouse liver tissue. HE staining was applied to observe pathological changes in mouse liver tissue. ELISA method was applied to detect serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC) and triglycerides (TG) in mice. The expression of UBA52 protein in liver was detected by Western blot assay. HepG2 cells were treated with 0.5 mmol/L oleic acid to induce an in vitro metabolic dysfunction-associated steatotic liver disease model. HepG2 cells were randomly divide into the control group, the oleic acid induced group, the low concentration ERI group (ERI low group, 50 μmol/L ERI), the high concentration ERI group (ERI high group, 100 μmol/L ERI), the high concentration ERI+si-NC group (ERI high+si-NC group, 100 μmol/L ERI+transfected with si-NC) and the high concentration ERI+si-UBA52 group (ERI high+si-UBA52 group, 100 μmol/L ERI+transfected with si-UBA52). Oil red O staining was applied to detect lipid deposition in HepG2 cells of each group. ELISA method was applied to detect the levels of TG, TC, SOD and MDA in HepG2 cells in each group. Immunoblotting was used to detect the expression levels of UBA52, p62 and autophagy related proteins in HepG2 cells. Results Compared with the normal group, serum levels of ALT, AST, LDL-C, TC, TG and the expression of UBA52 protein in liver tissue were increased in the model group (P<0.05), and the lipid deposition in liver increased, pathological damage was severe, and the proportion of lipid deposition area and non-alcoholic fatty liver disease (NAFLD) activity score were also increased (P<0.05). Changes in the corresponding indicators in the ERI-L group and the ERI-H group were opposite to those of the model group (P<0.05), and the ERI-H group was even lower (P<0.05). The lipid deposition in liver decreased and the pathological damage was alleviated. Compared with the control group, the levels of TG, TC, MDA, the proportion of lipid droplet area and the expression of UBA52 protein were increased in HepG2 cells of the oleic acid-induced group, while the levels of SOD, p62 and LC3Ⅱ/LC3Ⅰ decreased (P<0.05). Changes in the corresponding indicators of the low-concentration ERI group and the high-concentration ERI group were opposite to those of the oleic acid-induced group (P<0.05), and the therapeutic effect of ERI on metabolic dysfuntion-associated steatotic liver disease was enhanced after knocking down the expression of UBA52. Conclusion ERI may slow down the progression of metabolic dysfuntion-associated steatotic liver disease by down-regulating the expression of UBA52 at both in vivo and in vitro levels.

Key words: autophagy, oxidative stress, metabolic dysfunction-associated steatotic liver disease, eriodictyol, ubiquitin A 52

CLC Number:

R575

Figures/Tables 8

References 24

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组别	ALT/（U/L）		AST/（U/L）
Normal组	22.38±2.17		104.72±8.82
Model组	79.58±6.20^a		158.12±12.64^a
ERI-L组	60.26±5.54^b		134.49±10.43^b
ERI-H组	34.69±3.05^bc		109.28±7.65^bc
F	380.091^＊＊		72.496^＊＊
组别	LDL-C/（mmol/L）	TC/（mmol/L）	TG/（mmol/L）
Normal组	0.31±0.03	3.04±0.22	0.96±0.07
Model组	0.98±0.10^a	6.32±0.56^a	1.72±0.15^a
ERI-L组	0.73±0.07^b	5.10±0.43^b	1.48±0.15^b
ERI-H组	0.45±0.05^bc	3.35±0.31^bc	1.10±0.10^bc
F	233.158^＊＊	177.107^＊＊	97.095^＊＊

组别	ALT/（U/L）		AST/（U/L）
Normal组	22.38±2.17		104.72±8.82
Model组	79.58±6.20^a		158.12±12.64^a
ERI-L组	60.26±5.54^b		134.49±10.43^b
ERI-H组	34.69±3.05^bc		109.28±7.65^bc
F	380.091^＊＊		72.496^＊＊
组别	LDL-C/（mmol/L）	TC/（mmol/L）	TG/（mmol/L）
Normal组	0.31±0.03	3.04±0.22	0.96±0.07
Model组	0.98±0.10^a	6.32±0.56^a	1.72±0.15^a
ERI-L组	0.73±0.07^b	5.10±0.43^b	1.48±0.15^b
ERI-H组	0.45±0.05^bc	3.35±0.31^bc	1.10±0.10^bc
F	233.158^＊＊	177.107^＊＊	97.095^＊＊

组别	TG（mmol/g）	TC（mmol/g）	SOD（U/mg）	MDA（μmol/g）
Control组	0.12±0.01	0.18±0.02	52.65±4.27	1.02±0.08
油酸诱导组	0.40±0.04^a	0.43±0.04^a	16.15±1.86^a	2.84±0.20^a
低浓度ERI组	0.29±0.03^b	0.35±0.03^b	27.22±2.36^b	2.38±0.20^b
高浓度ERI组	0.19±0.01^bc	0.27±0.02^bc	38.49±3.50^bc	1.71±0.17^bc
高浓度ERI+ si-NC组	0.19±0.02	0.26±0.03	39.02±3.88	1.69±0.17
高浓度ERI+ si-UBA52组	0.13±0.01^de	0.19±0.02^de	50.31±4.75^de	1.24±0.12^de
F	128.700^＊＊	72.000^＊＊	89.447^＊＊	107.098^＊＊

组别	TG（mmol/g）	TC（mmol/g）	SOD（U/mg）	MDA（μmol/g）
Control组	0.12±0.01	0.18±0.02	52.65±4.27	1.02±0.08
油酸诱导组	0.40±0.04^a	0.43±0.04^a	16.15±1.86^a	2.84±0.20^a
低浓度ERI组	0.29±0.03^b	0.35±0.03^b	27.22±2.36^b	2.38±0.20^b
高浓度ERI组	0.19±0.01^bc	0.27±0.02^bc	38.49±3.50^bc	1.71±0.17^bc
高浓度ERI+ si-NC组	0.19±0.02	0.26±0.03	39.02±3.88	1.69±0.17
高浓度ERI+ si-UBA52组	0.13±0.01^de	0.19±0.02^de	50.31±4.75^de	1.24±0.12^de
F	128.700^＊＊	72.000^＊＊	89.447^＊＊	107.098^＊＊

组别	UBA52	p62	LC3Ⅱ/LC3Ⅰ
Control组	0.20±0.02	0.93±0.09	0.89±0.09
油酸诱导组	0.86±0.09^a	0.23±0.02^a	0.26±0.03^a
低浓度ERI组	0.70±0.07^b	0.42±0.04^b	0.46±0.05^b
高浓度ERI组	0.48±0.05^bc	0.63±0.06^bc	0.68±0.07^bc
高浓度ERI+si-NC组	0.47±0.05	0.62±0.06	0.70±0.07
高浓度ERI+si-UBA52组	0.25±0.03^de	0.82±0.08^de	0.86±0.09^de
F	120.547^＊＊	99.489^＊＊	71.727^＊＊