Research on emodin improving high glucose-induced mitophagy impairment in hippocampal neurons

doi:10.11958/20252781

Abstract

Abstract:

Objective To investigate the protective effect of emodin, an active component of polygonum multiflorum, against mitochondrial autophagy dysfunction in hippocampal neurons under high-glucose conditions by inhibiting histone acetyltransferase 2A (KAT2A), and its potential molecular mechanism. Methods An in vitro model was established by inducing hippocampal neurons with high glucose. Following intervention with emodin, cell viability, lactate dehydrogenase (LDH) content, histone acetyltransferase (HAT) content and reactive oxygen species (ROS) levels were measured, and mitochondrial membrane potential changes were also detected. Western blot assay and RT-qPCR were employed to assess the expression of mitochondrial autophagy-related proteins, including microtubule-associated protein 1 light chain 3-Ⅱ(LC3-Ⅱ), P62, PTEN-induced prokinase 1 (PINK1), KAT2A and their corresponding mRNAs. Molecular docking analysis was conducted to evaluate the binding affinity between emodin and KAT2A. Results Emodin treatment significantly enhanced neuronal viability under high-glucose conditions, while reducing LDH leakage, HAT and intracellular ROS levels, and effectively mitigated mitochondrial membrane potential decline. Additionally, Western blot assay and RT-qPCR results demonstrated that after emodin intervention, the expression levels of KAT2A, P62 and PINK1 proteins and mRNA were significantly downregulated, while LC3-Ⅱ protein and mRNA expression were upregulated. Molecular docking results further revealed that emodin exhibited good binding activity with KAT2A, with a binding energy of -32.6 kJ/mol. Conclusion This study demonstrates that emodin can effectively alleviate high glucose-induced mitophagy dysfunction in hippocampal neurons and reduce the generation of ROS and LDH. The underlying mechanism may be associated with the inhibition of KAT2A activity.

Key words: emodin, histone acetyltransferases, hippocampus, neurons, mitochondria, autophagy

CLC Number:

R587.25

Figures/Tables 7

References 30

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基因名称	引物序列（5′→3′）	产物大小/bp
KAT2A	上游：TAGCTGTGAGCACCCTTTGG	107
	下游：TTCCTCTTCTCCCCTGGCAT	107
LC3-Ⅱ	上游：GATGTCCGACTTATTCGAGAGC	82
	下游：TTGAGCTGTAAGCGCCTTCTA	82
P62	上游：GGACCCATCTACAGAGGCTG	96
	下游：TTCTGGACTGCTGGTGCAAT	96
PINK1	上游：GAGTGGGACTCAGATGGCTG	92
	下游：GCACATTTGCAGCTAAGCGT	92
β-actin	上游：GTGACGTTGACATCCGTAAAGA	114
	下游：GTAACAGTCCGCCTAGAAGCAC	114

基因名称	引物序列（5′→3′）	产物大小/bp
KAT2A	上游：TAGCTGTGAGCACCCTTTGG	107
	下游：TTCCTCTTCTCCCCTGGCAT	107
LC3-Ⅱ	上游：GATGTCCGACTTATTCGAGAGC	82
	下游：TTGAGCTGTAAGCGCCTTCTA	82
P62	上游：GGACCCATCTACAGAGGCTG	96
	下游：TTCTGGACTGCTGGTGCAAT	96
PINK1	上游：GAGTGGGACTCAGATGGCTG	92
	下游：GCACATTTGCAGCTAAGCGT	92
β-actin	上游：GTGACGTTGACATCCGTAAAGA	114
	下游：GTAACAGTCCGCCTAGAAGCAC	114

组别	细胞活力/%	LDH（U/L）
对照组	100.00±2.79	0.313 4±0.004 6
高糖组	67.23±3.99^a	0.353 8±0.004 8^a
高糖组+大黄素组	84.06±4.06^ab	0.339 8±0.008 8^ab
F	204.400^＊＊	112.300^＊＊
组别	ROS		HAT（μg/L）
对照组	444.33±34.67		0.48±0.47
高糖组	1 227.33±157.67^a		1.31±0.02^a
高糖组+大黄素组	840.67±76.67^ab		0.88±0.03^ab
F	151.400^＊＊		5 101.000^＊＊

组别	细胞活力/%	LDH（U/L）
对照组	100.00±2.79	0.313 4±0.004 6
高糖组	67.23±3.99^a	0.353 8±0.004 8^a
高糖组+大黄素组	84.06±4.06^ab	0.339 8±0.008 8^ab
F	204.400^＊＊	112.300^＊＊
组别	ROS		HAT（μg/L）
对照组	444.33±34.67		0.48±0.47
高糖组	1 227.33±157.67^a		1.31±0.02^a
高糖组+大黄素组	840.67±76.67^ab		0.88±0.03^ab
F	151.400^＊＊		5 101.000^＊＊

组别		线粒体膜电位		KAT2A
对照组		1.84±0.17		1.00±0.00
高糖组		0.55±0.11^a		1.47±0.15^a
高糖组+大黄素组		1.20±0.07^ab		0.76±0.11^ab
F		92.810^＊＊		44.920^＊＊
组别	LC3-Ⅱ		P62		PINK1
对照组	1.00±0.00		1.00±0.00		1.00±0.00
高糖组	0.75±0.25		1.38±0.15		1.53±0.13^a
高糖组+大黄素组	1.43±0.28^b		0.76±0.37^b		0.73±0.21^b
F	10.010^＊		7.129^＊		25.180^＊＊