iTRAQ-based quantitative proteomics revealing the therapeutic mechanism of a medicinal and edible formula YH0618 in reducing doxorubicin-induced alopecia by targeting keratins and TGF-β/Smad3 pathway

Renkai Li; Mingxia Chen; Danxi Yan; Liang Chen; Mandi Lin; Bohui Deng; Likai Zhuang; Fei Gao; George Pak-Heng Leung; Jieshu You

doi:10.1016/j.heliyon.2024.e33051

iTRAQ-based quantitative proteomics revealing the therapeutic mechanism of a medicinal and edible formula YH0618 in reducing doxorubicin-induced alopecia by targeting keratins and TGF-β/Smad3 pathway

Heliyon. 2024 Jun 18;10(12):e33051. doi: 10.1016/j.heliyon.2024.e33051. eCollection 2024 Jun 30.

Authors

Renkai Li^{1

2}, Mingxia Chen¹, Danxi Yan¹, Liang Chen³, Mandi Lin⁴, Bohui Deng¹, Likai Zhuang¹, Fei Gao⁵, George Pak-Heng Leung², Jieshu You¹

Affiliations

¹ College of Pharmacy, Shenzhen Technology University, Room 704, Block A2, 3002 Lantian Road, Pingshan District, Shenzhen, Guangdong Province, China.
² Department of Pharmacology and Pharmacy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
³ School of Exercise and Health, Shanghai University of Sport, Shanghai, China.
⁴ Department of Radiotherapy, The First Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong Province, China.
⁵ College of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu, Sichuan Province, China.

Abstract

YH0618, a medicinal and edible formulation, has demonstrated the potential to alleviate doxorubicin-induced alopecia in animal studies and clinical trials. However, the mechanisms underlying its therapeutic effects remain unexplored. The objective of this study was to ascertain possible therapeutic targets of YH0618 in the treatment of doxorubicin-induced alopecia. The assessment of hair loss was conducted through the measurement of the proportion of the affected area and the examination of skin histology. Isobaric tags for relative and absolute quantification (iTRAQ) in quantitative proteomics was employed to discern proteins that exhibited variable expressions. The major proteins associated with doxorubicin-induced alopecia were identified using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis. The interaction network of the differentially expressed proteins was constructed using the STRING database and the Python software. The study analyzed a total of 3894 proteins extracted from the skin tissue of mice. Doxorubicin treatment resulted in the upregulation of 18 distinct proteins, whereas one differential protein was found to be downregulated. The above effects were reinstated after the administration of the YH0618 therapy. The bioinformatic study revealed that the identified proteins exhibited enrichment in many biological processes, including staphylococcus aureus infection, estrogen signaling route, pyruvate metabolism, chemical carcinogenesis, and PPAR signaling pathway. The results of Western blot revealed that the levels of keratin 81 (Krt81), keratin 34 (Krt34), keratin 33a (Krt33a), and Sma and MAD-related protein 3 (Smad3) were upregulated in response to doxorubicin treatment, and were attenuated by the administration of YH0618. These four proteins are likely to correlate with DOX-induced alopecia and serve as promising therapeutic targets for YH0618. This work presents significant insights and empirical evidence for comprehending the process underlying chemotherapy-induced alopecia, paving the way for exploring innovative therapeutic or preventive strategies employing herbal items.

Keywords: Alopecia; Doxorubicin; Herbal formula; Network pharmacology; Proteomics.