Page 137 - 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

PC-19

Sulfated polysaccharide from Antrodia cinnamomea mycelium cultured with

zinc sulfate stimulates M1 polarization of macrophages through AKT/mTOR

2
7
1,2
Zhi-Hu Lin, Mei-Kuang Lu, 2,4,5,6 Chia-Chuan Chang, Ai-Jung Tseng, Chi-Hsein
4
8,9
Chao, Chi-Hong Chao, Tung-Yi Lin* ,2,3,4

1 Department of ophthalmology, Taipei City Hospital, Taipei 103009, Taiwan
2 Institute of Traditional Medicine, National Yang Ming Chiao Tung University, Taipei
112304, Taiwan
3 Program in Molecular Medicine, National Yang-Ming University and Academia Sinica,
Taipei 112304, Taiwan
4 School of Chinese Medicine, National Yang Ming Chiao Tung University, Taipei 112304,
Taiwan
5 National Research Institute of Chinese Medicine, Ministry of Health and Welfare, Taipei
112108, Taiwan
6 Graduate Institute of Pharmacognosy, Taipei Medical University, Taipei 110497, Taiwan
7 School of Pharmacy, College of Medicine, National Taiwan University, Taipei 100025,
Taiwan
8 Department of Biological Science and Technology, National Yang Ming Chiao Tung
University, Hsinchu 300193, Taiwan
9 Center For Intelligent Drug Systems and Smart Bio-devices (IDS2B), National Yang Ming
Chiao Tung University, Hsinchu 300193, Taiwan
* E-mail: tylin99@nycu.edu.tw

Abstract
Sulfated polysaccharides (Ac-SPSs) from Antrodia cinnamomea are valued for their anti-
cancer and immune-modulating activities. However, their naturally low yield limits practical
application. Previous studies have suggested that trace minerals can influence fungal
metabolism. In this study, we examined whether zinc sulfate (ZnSO₄) could enhance Ac-SPS
production and investigated the most active fraction for its role in macrophage polarization and
suppression of lung cancer cell growth. When A. cinnamomea mycelia were treated with ZnSO₄
(1, 10, and 100 µM), Ac-SPS yield increased by approximately 20–30%. The 10 µM treatment
consistently produced a medium-molecular-weight fraction (~200 kDa, ASZ-10) with the most
pronounced bioactivity. ASZ-10 directly inhibited the proliferation of A549 lung cancer cells.
Although it did not reduce LPS-induced inflammation, it promoted M1 macrophage
polarization, enhancing the secretion of nitric oxide, TNF-α, IL-6, and M1 surface markers
(CD86, CD80, CD64, MHCII) without affecting the M2 marker CD206. In M-CSF-stimulated
bone marrow-derived macrophages, ASZ-10 also increased CD80⁺ and CD86⁺ expression. This
effect depended on AKT/mTOR signaling, as pathway inhibition with LY294002 or rapamycin
diminished M1 induction. Notably, co-culturing macrophages with lung cancer cells under
ASZ-10 stimulation further reduced tumor viability. These findings suggest that ZnSO₄
supplementation is a straightforward approach to increase Ac-SPS production. ASZ-10 exerts
dual anti-cancer effects, direct inhibition of cancer cells and macrophage-mediated cytotoxicity,
supporting its potential as a functional supplement or adjuvant in lung cancer therapy,
particularly in strategies utilizing immune activation to improve treatment outcomes.

Keywords: Sulfated polysaccharides; Antrodia cinnamomea; Zinc sulfate; Biochemical
characterization; Macrophage