Page 221 - 2025中醫藥與天然藥物聯合學術研討會-中醫藥與天然藥物的挑戰Ｘ機遇與未來大會手冊

PP-39

Helminthostachys zeylanica extracts mitigate inflammation and senescence

via the regulation of p53-p21/CDKN1A axis in skin keratinocytes

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Pei-Xuan Wu, Hui-Ching Tseng, Yin-Chen Chen, Hsi-Lung Hsieh* ,1,2,3

1 Graduate Institute of Health Industry Technology, College of Human Ecology, Chang Gung
University of Science and Technology, Taoyuan, Taiwan
2 Center for Drug Research and Development, College of Human Ecology, Chang Gung
University of Science and Technology, Taoyuan 33303, Taiwan
3 Department of Neurology, Chang Gung Memorial Hospital, Taoyuan, Taiwan
* E-mail: hlhsieh@mail.cgust.edu.tw

Abstract
Skin aging is closely associated with chronic inflammation and cellular senescence, during
which skin cells undergo cell cycle arrest through activation of the p53-p21/CDKN1A pathway
and acquire a senescence-associated secretory phenotype (SASP). Previous studies have
demonstrated that Helminthostachys zeylanica (L.) possesses anti-inflammatory properties,
reduces oxidative stress, and alleviates liver and lung injuries. Ugonins and ugonstibens are
major compounds enriched in H. zeylanica extracts. Therefore, we aimed to investigate whether
H. zeylanica (HZ) extracts and their index constituents can mitigate skin inflammation and
aging. Our previous findings revealed that D-galactose (D-gal) induces senescence in skin
keratinocytes (HaCaT cells), characterized by increased expression of SASP factors (IL-1β,
TNF-α, and IL-6), upregulation of COX-2, and elevated senescence-associated β-galactosidase
(SA-β-gal) activity. Here, we further demonstrate that D-gal regulates the senescence marker
p21/CDKN1A, providing a basis to evaluate the effects of HZ extracts and their active
compounds on skin inflammation and aging. D-gal induced senescence in HaCaT cells,
evidenced by a concentration-dependent increase in p21/CDKN1A protein, accompanied by
upregulation of CDKN1A mRNA. Intriguingly, total p53 protein decreased in a concentration-
dependent manner, whereas p53 phosphorylation increased, a pattern consistent with activation-
associated turnover of p53 that drives p21 transcription. Mechanistically, treatment with the
ethyl acetate extract of H. zeylanica (HZ-EA) and its key constituents significantly suppressed
D-gal-induced p53 phosphorylation and reduced both p21 protein and CDKN1A transcript
levels, and diminished SA-β-gal activity. HZ-EA and its constituents attenuated the D-gal-
induced inflammatory response, lowering IL-1β, IL-6, and COX-2 expression. Collectively, HZ
extracts and their active constituents reduce skin inflammation and cellular aging through
suppression of the COX-2/p53/p21 axis and relief of cell-cycle arrest, supporting their promise
as natural skincare candidates.

Keywords: Helminthostachys zeylanica; Cellular senescence; Skin inflammation;
p21/CDKN1A