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Humulus lupulus L. extract and its active constituent xanthohumol attenuate oxidative stress and nerve injury induced by iron overload via activating AKT/GSK3β and Nrf2/NQO1 pathways

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Abstract

Hops, the dried female clusters from Humulus lupulus L., have traditionally been used as folk medicines for treating insomnia, neuralgia, and menopausal disorders. However, its pharmacological action on iron overload induced nerve damage has not been investigated. This study aims to evaluate the protective effects of hops extract (HLE) and its active constituent xanthohumol (XAN) on nerve injury induced by iron overload in vivo and in vitro, and to explore its underlying mechanism. The results showed that HLE and XAN significantly improved the memory impairment of iron overload mice, mainly manifested as shortened latency time, increased crossing platform times and spontaneous alternation ratio, and increased the expression of related proteins. Additionally, HLE and XAN significantly increased superoxide dismutase (SOD) and glutathione peroxidase (GSH-PX) activities, and remarkably decreased malondialdehyde (MDA) level in hippocampus. Also, HLE and XAN apparently reduced reactive oxygen species (ROS) content of PC12 cells induced by iron dextran (ID), and improved the oxidative stress level. Moreover, HLE and XAN significantly upregulated the expression of nuclear factor E2-related factor (Nrf2), NAD(P)H quinone oxidoreductase (NQO1), heme oxygenase-1 (HO-1), SOD, phosphorylated AKT (p-AKT), and phosphorylated GSK3β (p-GSK3β) both in hippocampus and PC12 cells. These findings demonstrated the protective effect of HLE and XAN against iron-induced memory impairment, which is attributed to its antioxidant profile by activation of AKT/GSK3β and Nrf2/NQO1 pathways. Also, it was suggested that hops could be a potential candidate for iron overload-related neurological diseases treatment.

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Abbreviations

HLE:

Hops extract

XAN:

Xanthohumol

ID:

Iron dextran

MWM:

Morris water maze

BDNF:

Brain-derived neurotrophic factor

ERK:

Extracellular regulated protein kinases

CREB:

cAMP-response element binding protein

AKT:

Protein kinase B

GSK-3β :

Glycogen synthase kinase-3β

Nrf2:

Nuclear factor E2-related factor

SOD:

Superoxide dismutase

GSH-PX:

Glutathione peroxidase

MDA:

Malondialdehyde

NQO1:

NAD(P)H:quinone oxidoreductase 1

HO-1:

Heme oxygenase-1

Aβ :

Amyloid β-protein

ROS:

Reactive oxygen species

H-A:

Huperzine-A

PRRSV:

Porcine reproductive and respiratory syndrome virus

SAMP8:

Senescence-accelerated prone mice

BCA:

Bicinchoninic acid

LDH:

Lactate dehydrogenase

SDS-PAGE:

Sodium dodecylsulfate polyacrylamide gel electrophoresis

PVDF:

Polyvinylidine difluoride

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

ECL:

Enhanced chemiluminescence

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (82174079, 82004015), and Project of Science and Technology Commission of Shanghai Municipality (21S21902600).

Funding

National Natural Science Foundation of China, 82174079, Xin Hailiang, 82004015, Xia Tian-Shuang, Project of Science and Technology Commission of Shanghai Municipality, 21S21902600, Xin Hailiang.

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Author notes

  1. Sun Xiao-Lei and Xia Tian-Shuang have contributed equally to this work and are considered to be co-first authors.

Authors and Affiliations

  1. Department of Pharmacognosy, School of Pharmacy, Naval Medical University, 325 Guohe Road, Shanghai, 200433, China

    Sun Xiao-Lei, Xia Tian-Shuang, Jiang Yi-Ping, Han Ting & Xin Hai-Liang

  2. School of Pharmacy, Shandong University of Traditional Chinese Medicine, 4655 Daxue Road, Jinan, 250355, China

    Sun Xiao-Lei & Xu Ling-Chuan

  3. Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou, 310007, Zhejiang, China

    Wang Na-Ni

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  1. Sun Xiao-Lei
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Contributions

XH-L, SX-L, and XT-S designed the experiments; SX-L performed the experiments; SX-L and XT-S analyzed the data, wrote and revised the manuscript; JY-P helped to organize figures, WN-N contributed to the documents download, XL-C, HT and XH-L reviewed the manuscript.

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Correspondence to Xu Ling-Chuan, Han Ting or Xin Hai-Liang.

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Appendix 1

Appendix 1

HPLC profile of Humulus lupulus L. (9-xanthohumol, 13-cohumulone, 14-humulone, 15-adhumulone, 17-colupulone, 18-lupulone, 19-adlupulone)

figure a

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Xiao-Lei, S., Tian-Shuang, X., Yi-Ping, J. et al. Humulus lupulus L. extract and its active constituent xanthohumol attenuate oxidative stress and nerve injury induced by iron overload via activating AKT/GSK3β and Nrf2/NQO1 pathways. J Nat Med 77, 12–27 (2023). https://doi.org/10.1007/s11418-022-01642-1

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