Identification of anti-fibrotic and pro-apoptotic bioactive compounds from Ganoderma formosanum and their possible mechanisms in modulating TGF-β1-induced lung fibrosis

Kuan-Chen Cheng; Patrick Chun Theng Chong; Chen-Che Hsieh; Yu-Te Lin; Chih-Hung Ye; Darin Khumsupan; Jheng-Jhe Lu; Wei-Chieh Yu; Kai-Wen Cheng; Kah Yi Yap; Weng Si Kou; Meng-Tsung Cheng; Cheng-Chih Hsu; Lee-Yan Sheen; Shin-Ping Lin; An-Chi Wei; Shu-Han Yu

doi:10.1016/j.jep.2024.118008

Identification of anti-fibrotic and pro-apoptotic bioactive compounds from Ganoderma formosanum and their possible mechanisms in modulating TGF-β1-induced lung fibrosis

J Ethnopharmacol. 2024 Jun 12:327:118008. doi: 10.1016/j.jep.2024.118008. Epub 2024 Mar 6.

Authors

Kuan-Chen Cheng¹, Patrick Chun Theng Chong², Chen-Che Hsieh², Yu-Te Lin³, Chih-Hung Ye², Darin Khumsupan², Jheng-Jhe Lu², Wei-Chieh Yu², Kai-Wen Cheng⁴, Kah Yi Yap², Weng Si Kou², Meng-Tsung Cheng⁵, Cheng-Chih Hsu⁶, Lee-Yan Sheen⁷, Shin-Ping Lin⁸, An-Chi Wei³, Shu-Han Yu⁹

Affiliations

¹ Institute of Biotechnology, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C; Institute of Food Science Technology, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C; Department of Optometry, Asia University, No. 500, Lioufeng Rd., Wufeng, Taichung, Taiwan. R.O.C; Department of Medical Research, China Medical University Hospital, China Medical University, No. 91, Hsueh-Shih Rd., Taichung, Taiwan. R.O.C.
² Institute of Biotechnology, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C.
³ Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei 10617, Taiwan. R.O.C.
⁴ Department of Chemistry, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C.
⁵ School of Pharmacy, College of Medicine, National Taiwan University, No.33, Linsen S. Rd., Taipei, 100025, Taiwan. R.O.C.
⁶ Department of Chemistry, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C; Leeuwenhoek Laboratories Co. Ltd., No. 71, Fanglan Rd, Taipei, 106038, Taiwan. R.O.C.
⁷ Institute of Food Science Technology, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C.
⁸ School of Food Safety, Taipei Medical University, No. 250, Wu-Hsing Street, Taipei, Taiwan. R.O.C.
⁹ Institute of Biotechnology, National Taiwan University, No. 1, Sec. 4, Roosevelt Rd., Taipei, Taiwan. R.O.C. Electronic address: shuhanyu@ntu.edu.tw.

PMID: 38458343
DOI: 10.1016/j.jep.2024.118008

Abstract

Ethnopharmacological relevance: The Compendium of Materia Medica and the Classic of Materia Medica, the two most prominent records of traditional Chinese medicine, documented the therapeutic benefits of Ganoderma sinense particularly in addressing pulmonary-related ailments. Ganoderma formosanum, an indigenous subspecies of G. sinense from Taiwan, has demonstrated the same therapeutic properties.

Aim of the study: The aim of this study is to identify bioactive compounds and evaluate the potential of G. formosanum extracts as a novel treatment to alleviate pulmonary fibrosis (PF). Using an in-house drug screening platform, two-stage screening was performed to determine their anti-fibrotic efficacy.

Methods and materials: G. formosanum was fractionated into four partitions by solvents of different polarities. To determine their antifibrotic and pro-apoptotic properties, the fractions were analyzed using two TGF-β1-induced pulmonary fibrosis cell models (NIH-3T3) and human pulmonary fibroblast cell lines, immunoblot, qRT-PCR, and annexin V assays. Subsequently, transcriptomic analysis was conducted to validate the findings and explore possible molecular pathways. The identification of potential bioactive compounds was achieved through UHPLC-MS/MS analysis, while molecular interaction study was investigated by multiple ligands docking and molecular dynamic simulations.

Results: The ethyl acetate fraction (EAF) extracted from G. formosanum demonstrated substantial anti-fibrotic and pro-apoptotic effects on TGF-β1-induced fibrotic models. Moreover, the EAF exhibited no discernible cytotoxicity. Untargeted UHPLC-MS/MS analysis identified potential bioactive compounds in EAF, including stearic acid, palmitic acid, and pentadecanoic acid. Multiple ligands docking and molecular dynamic simulations further confirmed that those bioactive compounds possess the ability to inhibit TGF-β receptor 1.

Conclusion: Potential bioactive compounds in G. formosanum were successfully extracted and identified in the EAF, whose anti-fibrotic and pro-apoptotic properties could potentially modulate pulmonary fibrosis. This finding not only highlights the EAF's potential as a promising therapeutic candidate to treat pulmonary fibrosis, but it also elucidates how Ganoderma confers pulmonary health benefits as described in the ancient texts.

Keywords: Anti-fibrosis; Ganoderma formosanum; Lung Protection; Pro-apoptosis; Pulmonary fibrosis; Transforming growth Factor-β1.

MeSH terms

Fibrosis
Ganoderma*
Humans
Lung
Materia Medica* / pharmacology
Pulmonary Fibrosis* / chemically induced
Pulmonary Fibrosis* / drug therapy
Pulmonary Fibrosis* / metabolism
Tandem Mass Spectrometry
Transforming Growth Factor beta1 / metabolism

Substances

Transforming Growth Factor beta1
Materia Medica

Supplementary concepts

Ganoderma formosanum