Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Jul 7;26(2):405.
doi: 10.3892/etm.2023.12104. eCollection 2023 Aug.

Ethyl acetate extract of Gastrodia elata protects Caenorhabditis elegans from oxidative stress and amyloid β peptide toxicity

Xiongfei Shi  1 , Xingzhi Yu  1 , Liping Yang  1 , Xiaohua Duan  1
Affiliations

Ethyl acetate extract of Gastrodia elata protects Caenorhabditis elegans from oxidative stress and amyloid β peptide toxicity

Xiongfei Shi et al. Exp Ther Med. .

Abstract

Gastrodia elata Blume is a traditional Chinese medicine with a long history, which has numerous pharmacological activities, such as anti-inflammation, anti-oxidation and protection of nerves. The present study investigated the regulatory effect of ethyl acetate extract of Gastrodia elata (EEGE) on the β-amyloid (Aβ) toxicity of Caenorhabditis elegans (C. elegans). First, the main components of EEGE were analyzed using high-performance liquid chromatography, and the total phenols, total flavonoids and total antioxidant capacity of EEGE were determined. Next, the regulation effect of EEGE on Aβ-induced toxicity of C. elegans was evaluated through experiments on nematode paralysis, lifespan, oxidative and heat stress, locomotor ability, reproductive ability, reactive oxygen species (ROS) level, Aβ aggregation test, malondialdehyde (MDA) level, catalase (CAT) activity and superoxide dismutase (SOD) activity. Finally, the mechanism of EEGE was elucidated using RNA sequencing (RNA-Seq) and the expression levels of related genes were verified using quantitative PCR. The present study revealed that the main components of EEGE included phosphorylated (p)-hydroxybenzyl alcohol, p-hydroxybenzaldehyde and 4,4'-dihydroxydiphenylmethane, possessing strong in vitro free radical scavenging and reducing abilities. In addition, after the intervention of EEGE, the paralysis of nematodes could be delayed, the survival time of the nematodes was prolonged, the survival rate of the nematodes under stress (high temperature and oxidation) conditions was improved, the activity capacity and reproductive capacity of the nematodes were improved, the activities of SOD and CAT were improved and the levels of ROS and MDA were reduced. Notably, EEGE directly inhibited Aβ plaque aggregation in nematodes. RNA-Seq analysis showed that EEGE regulated metabolism and longevity-related genes, and these genes were regulated by the insulin/IGF-1 signaling (IIS) pathway. Therefore, the present study hypothesized that the regulatory mechanism of EEGE was significantly related to the IIS pathway. The present research results demonstrated that the protective effect of EEGE on transgenic C. elegans was to reduce Aβ protein aggregation, improve the in vivo antioxidant level, effectively remove free radicals and to regulate the expression of genes related to IIS pathway, thereby reducing Aβ-induced toxicity and delaying nematode paralysis.

Keywords: Alzheimer's disease; Gastrodia elata Blume; RNA sequencing; oxidative stress; β-amyloid.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Composition analysis of EEGE. (A) Chromatogram of EEGE. (B) Chromatogram of standard. (C) Chemical structure of p-hydroxybenzyl alcohol, p-hydroxybenzaldehyde and 4,4'-dihydroxydiphenylmethane. EEGE, ethyl acetate extract of Gastrodia elata; p-, phosphorylated.
Figure 2
Figure 2
Effect of EEGE on C. elegans. Effects of EEGE on (A) paralysis, (B) life span, (C) heat stress, (D) Juglone-induced oxidative stress, (E) locomotor ability and (F) reproductive capacity of C. elegans. *P<0.05 and **P<0.01 vs. the Control. EEGE, Ethyl acetate extract of Gastrodia elata.
Figure 3
Figure 3
EEGE decreases ROS and Aβ accumulation in C. elegans. (A) ROS accumulation in C. elegans. (B) ROS fluorescence value per unit area. (C) Effect of EEGE on Aβ aggregation in N2 and CL2006 C. elegans. (D) Deposition of Aβ in C. elegans. The white light and fluorescence images of ROS in N2 C. elegans were observed under a fluorescence microscope (magnification, x100). The fluorescence images were observed in N2 and CL2006 C. elegans after Thioflavin S staining by a laser scanning confocal microscope (magnification, x200). **P<0.01 vs. the Control. EEGE, Ethyl acetate extract of Gastrodia elata; Aβ, β-amyloid; ROS, reactive oxygen species.
Figure 4
Figure 4
Effects of EEGE on antioxidant enzymes and ROS levels in C. elegans. (A) CAT activity. (B) SOD activity. (C) MDA content. (D) Continuous changes of ROS fluorescence in C. elegans. **P<0.01 vs. the Control. ROS, reactive oxygen species; SOD, superoxide dismutase; MDA, malondialdehyde; CAT, catalase; EEGE, Ethyl acetate extract of Gastrodia elata.
Figure 5
Figure 5
Distribution and enrichment analysis of DEGs of RNA-sequencing. (A) Volcano map of DEGs. Red dots represent upregulated genes, green dots downregulated genes and gray dots represent genes with no significant differences. (B) GO analysis for DEGs. (C) KEGG analysis for DEGs. DEGs, differentially expressed genes; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.
Figure 6
Figure 6
The result verification of RNA-Seq. (A) Validation of up-regulated genes in DEGs. (B) Validation of down-regulated genes in DEGs. **P<0.0 vs. the Control. RNA-Seq, RNA Sequencing; EEGE, Ethyl acetate extract of Gastrodia elata.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Lane CA, Hardy J, Schott JM. Alzheimer's disease. Eur J Neurol. 2018;25:59–70. doi: 10.1111/ene.13439. - DOI - PubMed
    1. De Strooper B, Karran E. The cellular phase of Alzheimer's disease. Cell. 2016;164:603–615. doi: 10.1016/j.cell.2015.12.056. - DOI - PubMed
    1. Mohsenzadegan M, Mirshafiey A. The immunopathogenic role of reactive oxygen species in Alzheimer disease. Iran J Allergy Asthma Immunol. 2012;11:203–216. - PubMed
    1. Alafuzoff I, Pikkarainen M, Arzberger T, Thal DR, Al-Sarraj S, Bell J, Bodi I, Budka H, Capetillo-Zarate E, Ferrer I, et al. Inter-laboratory comparison of neuropathological assessments of beta-amyloid protein: A study of the BrainNet Europe consortium. Acta Neuropathol. 2008;115:533–546. doi: 10.1007/s00401-008-0358-2. - DOI - PubMed
    1. Zhang X, Kang X, Du L, Zhang L, Huang Y, Wang J, Wang S, Chang Y, Liu Y, Zhao Y. Tanshinone IIA loaded chitosan nanoparticles decrease toxicity of β-amyloid peptide in a Caenorhabditis elegans model of Alzheimer's disease. Free Radical BiolMed. 2022;193:81–94. doi: 10.1016/j.freeradbiomed.2022.09.030. - DOI - PubMed

Grants and funding

Funding: The present study was supported by the National Natural Science Foundation of China (grant no. 81960733), the Open Project of Yunnan Key Laboratory of Dai and Yi Medicines (grant no. 202210ZD2206), the Xingdian Talent Support Program - Special for Young Talent (grant no. XDYC-QNRC-2022-0284), the National Administration of Traditional Chinese Medicine High-level Key Discipline Construction Project ‘Dai Medicine’ and ‘Dai Pharmacy’.