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
. 2021 Oct;11(10):2705-2714.
doi: 10.1002/2211-5463.13263. Epub 2021 Aug 27.

Cordycepin inhibits cell senescence by ameliorating lysosomal dysfunction and inducing autophagy through the AMPK and mTOR-p70S6K pathway

Affiliations

Cordycepin inhibits cell senescence by ameliorating lysosomal dysfunction and inducing autophagy through the AMPK and mTOR-p70S6K pathway

Shi Qi Zuo et al. FEBS Open Bio. 2021 Oct.

Abstract

Cell senescence is closely related to autophagy. In this article, we identified a natural nucleoside analogue, cordycepin, that has the ability to significantly improve lysosomal function, enhance the activity of the lysosomal representative protease cathepsin B (CTSB), and promote the expression of the functional protein lysosomal-associated membrane protein 2 (LAMP2) on the lysosomal membrane. Cordycepin then restores the damaged autophagy level of aging cells by activating the classic AMPK and mTOR-p70S6K signaling pathways, thus inhibiting cell senescence in an H2 O2 -induced stress-induced premature senescence (SIPS) cell model. This study provides new theoretical support for the further development of cordycepin and clinical antiaging drugs to inhibit cell senescence and suggests that the regulatory mechanisms of lysosomes in senescent cells should be considered when treating age-related diseases.

Keywords: AMPK signaling pathways; autophagy; cell senescence; cordycepin; lysosomal function; lysosomal protease.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Cordycepin delays cell senescence. (A) NIH3T3 cells were stained using a SA‐β‐gal assay kit after treatment with 400 μm H2O2 for 72 h, and representative images were selected from three independent tests. Scale bars = 20 μm. (B) Western blot (WB) image of aging marker proteins P53, P21, and P16 in NIH3T3 cells after hydrogen peroxide treatment at different concentrations. (C) DCFH‐DA fluorescence was used to measure the amounts of intracellular reactive oxygen species (ROS). Scale bars = 100 μm. (D) ELISA was used to detect different levels of pro‐inflammatory factors in the control group, senescence group, and cordycepin treatment group. (E) The immunofluorescence images show the changes in the senescence marker protein P53 in the control group, the senescence group, and the cordycepin treatment group. Scale bars = 50 μm. Representative images of three independent experiments are presented. Data were analyzed using Student’s t‐test and are presented as the mean ± SD. *P < 0.05, **P < 0.01. The following concentrations were used in the treatments depicted in C, D, and E: H2O2 (400 μm) and cordycepin (40 μm).
Fig. 2
Fig. 2
Cordycepin promotes autophagy in senescent cells. (A) Western blot analysis of autophagy‐related protein levels in cordycepin‐ and H2O2‐treated NIH3T3 cells. (B) Immunofluorescence images show the changes in LC3‐positive staining distribution in the control group, the senescent group, and the cordycepin group. The representative images of three independent experiments are presented. Scale bars = 20 μm.
Fig. 3
Fig. 3
Cordycepin affects AMPK and mTOR–p70S6K autophagy access to delay cellular senescence. (A) WB analysis of mammalian target of rapamycin (mTOR), p70S6 kinase (p70S6K), p‐p70S6K, adenosine 5′‐monophosphate‐activated protein kinase (AMPK), and p‐AMPK expression in control and senescent fibroblasts after treatment with 40 μm cordycepin for 48 h. (B) WB images of autophagy, mTOR, AMPK, and p70S6K protein levels in senescent cells treated with cordycepin and compound C (C.C). (C) WB images of autophagy and aging protein levels in senescent cells treated with cordycepin and C.C. (D) ELISA image of expression of different senescence‐associated secretory phenotypes (SASP) under treatment with 400 μm hydrogen peroxide. Data were analyzed using Student’s t‐test and are presented as the mean ± SD. *P < 0.05, **P < 0.01. (E) Immunofluorescence shows the changes in LC3 fluorescence staining in the senescent group and after the addition of cordycepin and C.C, and the representative images of three independent experiments are shown. Scale bars = 20 μm.
Fig. 4
Fig. 4
Cordycepin ameliorated lysosomal dysfunction in senescent cells. (A) Acid phosphatase activity was detected after H2O2 and cordycepin treatment. The data were analyzed using Student’s t‐test and are shown as the mean ± SD. *P < 0.05. (B) The control group and senescent fibroblasts were treated with 40 μm cordycepin for 48 h, and the WB indicates the expression of lysosomal‐associated membrane protein 2 (LAMP2) and representative enzyme cathepsin B (CTSB) on the lysosomal membrane. (C) The lysosomal status was probed with a LysoTracker Red DND‐99 kit after senescent cells were treated with cordycepin, and representative images of three independent replicates are shown. Scale bars = 10 μm. (D) WB detected the expression of senescence‐related protein as well as LAMP2 and CTSB after treatment with cordycepin alone and with NH4Cl. Data were analyzed using Student’s t‐test and are presented as the means ± SD. *P < 0.05, **P < 0.01.

Similar articles

Cited by

References

    1. Kuilman T, Michaloglou C, Mooi WJ and Peeper DS (2010) The essence of senescence. Genes Dev 24, 2463–2479. - PMC - PubMed
    1. Hernandez‐Segura A, Nehme J and Demaria M (2018) Hallmarks of cellular senescence. Trends Cell Biol 28, 436–453. - PubMed
    1. Manosroi J, Chankhampan C, Kumguan K, Manosroi W and Manosroi A (2014) In vitroanti‐aging activities of extracts from leaves of Ma Kiang (Cleistocalyx nervosum var. paniala). Pharm Biol 53, 862–869. - PubMed
    1. Gewirtz DA (2013) Autophagy and senescence: a partnership in search of definition. Autophagy 9, 808–812. - PMC - PubMed
    1. Tai H, Wang Z, Gong H, Han X, Zhou J, Wang X, Wei X, Ding Y, Huang N, Qin Jet al. (2017) Autophagy impairment with lysosomal and mitochondrial dysfunction is an important characteristic of oxidative stress‐induced senescence. Autophagy 13, 99–113. - PMC - PubMed

Publication types

MeSH terms