肝豆灵片通过调控PKCβⅡ/ACSL4/ALOX5信号通路改善肝豆状核变性铁死亡的机制

doi:10.3976/j.issn.1002-4026.20230170

Abstract

Abstract:

This study investigates the effects of Gandouling (GDL) tablets on ferroptosis in hepatolenticular degeneration in TX mice and their mechanism of action on the ferroptosis of HT22 cells induced by CuCl₂, based on the PKCβII/ACSL4/ ALOX5 signaling pathway. TX mice were divided into five groups: control, model, GDL tablet, Fer-1, and Glutathione. HT22 cells were also divided into five groups: control, model, GDL tablet, Fer-1, and GDL tablet + Fer-1. Hematoxylin and eosin staining was used to detect the pathological changes in the hippocampus tissues of the mice. Western blotting was used to detect the expression of PKCβⅡ, ACSL4, and ALOX5 in the hippocampus tissues and HT22 cells of the mice, as well as the expression of SLC7A11 and GPX4 in HT22 cells. The content of Fe²⁺ in the hippocampus tissues of the mice was detected via microassay. The levels of SOD, MDA, and GSH-Px in HT22 cells were detected by microplate assay. Finally, the expression of PKCβⅡ, ACSL4, and ALOX5 mRNA in HT22 cells was detected by quantitative real-time polymerase chain reaction. Compared with the control group, the hippocampus tissues of mice in the model group showed clear damage; the protein expression of PKCβⅡ, ACSL4, and ALOX5 showed a clear increase; the protein expression of SLC7A11 and GPX4 decreased significantly; and Fe²⁺content increased significantly (P<0.05). Compared with the model group, the pathological damage to hippocampus tissues showed improvements in the GDL tablet, Fer-1, and Glutathione group with the effects being noticeable in the GDL tablet group. It was possible to inhibit ferroptosis of HT22 cells in the GDL tablet and Fer-1 group and significantly lower their expression of PKCβⅡ, ACSL4, and ALOX5 protein and mRNA in comparison to the model group(P<0.05). The MDA contentalso decreased significantly (P<0.05) while the SOD activity and the GSH-Px content increased significantly(P<0.05). Thus, GDL tablets can inhibit ferroptosis in hippocampus tissues of TX mice andinhibit ferroptosis induced by CuCl₂ in HT22 cells. Moreover, the ferroptosis mechanism may be related to the down-regulation of the PKCβⅡ, ACSL4, and ALOX5 signaling pathway and the attenuation of intracellular lipid peroxidation.

Key words: Gandouling tablets, hepatolenticular degeneration, PKCβⅡ/ACSL4/ALOX5 signaling pathway, ferroptosis, mechanism of action

CLC Number:

R285

WU Bojin, DONG Ting, WEN Yuya, TIAN Liwei, ZHAO Chenling. The mechanism by which Gandouling tablets improve ferroptosis in hepatolenticular degeneration through PKCβⅡ/ACSL4/ALOX5 signaling pathway regulation[J].Shandong Science, 2024, 37(4): 34-44.

Figures/Tables 10

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References 18

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基因名称	引物序列	产物长度/bp
PKCβⅡ	上游5'-ATCGCTGCTGTATGGACTTATTCAC-3'	130
	下游5'-AACACTTGATGGTCTTGGTCTTCTG-3'
ACSL4	上游5'-CCTGAGGGGCTTGAAATTCAC-3'	89
	下游5'-GTTGGTCTACTTGGAGGAACG-3'
ALOX5	上游5'-ACGCATCTGGTGTCTGAGGTG-3'	318
	下游5'-GAGCAGTCCATCATCACGATAGAAG-3'
GPX4	上游5'TGTGCATCCCGCGATGATT-3'	97
	下游5'CCCTGTACTTATCCAGGCAGA-3'
SLC7A11	上游5'GCATTCCCAGGGGCTAACAT-3'	282
	下游5'AATTTCTCCCATGCGGGTGT-3'
GADPH	上游5'GTGTTCCTACCCCCAATGTG-3'	215
	下游5'GTCATTGAGAGCAATGCCAG-3'

The mechanism by which Gandouling tablets improve ferroptosis in hepatolenticular degeneration through PKCβⅡ/ACSL4/ALOX5 signaling pathway regulation

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