Combined Physicochemical and Transcriptomic Analyses Reveal the Effect of Black Wolfberry Ferment on Alopecia

Ping Yu¹, Xiangbo Min¹, Lina Li¹, Yu Wang² and Di Zhao^1,

¹Jiangxi Renren Health Microecological Technology Co.,Ltd, Yichun, Jiangxi, China

²College of Food Science and Technology, Bohai University, Jinzhou, Liaoning, China

Cite this paper:
Ping Yu, Xiangbo Min, Lina Li, Yu Wang and Di Zhao. Combined Physicochemical and Transcriptomic Analyses Reveal the Effect of Black Wolfberry Ferment on Alopecia. Journal of Food and Nutrition Research. 2025; 13(5):195-205. doi: 10.12691/jfnr-13-5-1

Abstract

This study aimed to evaluate the therapeutic effect of black wolfberry (Lycium ruthenicum Murr.) ferment on mice with androgenetic alopecia (AGA). Mice were arbitrarily clustered into four groups: control (C), model (M), low-dosage ferment (LF), and high-dosage ferment (HF) groups. Mice in the M, LF, and HF groups had AGA induced for 16 d. Then, the C group was treated with normal saline for 16 d; and LF and HF mice received daily doses of 2.5 and 25 mL/kg body weight of black wolfberry ferment, respectively. The results indicate that black wolfberry ferment increased the weight of newborn hair on mice with AGA. It also enhanced the number of hair follicles, terminal hairs, and vellus hairs, and increased the dermal and hypodermal thickness, as verified by the proliferation of hair follicles expressing Ki-67 and hair follicle stem cell marker Sox9. A total of 1,636 genes were selected as necessary genes using differential gene expression and weighted gene co-expression network analysis. The KEGG analysis showed that the promotion of hair regrowth was most likely associated with the PI3K/AKT, NF-kappa B, and MAPK signaling pathways. Black wolfberry ferment can be used as a therapeutic agent for AGA, and the potential genes and pathways involved in its action were identified using transcriptomics.

Keywords:
androgenic alopecia Lycium ruthenicum Murr. ferment transcriptomics enrichment analysis

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References:

[1]	Z. Liu, Q. Shu, L. Wang, M. Yu, Y. Hu, H. Zhang, Y. Tao, and Y. Shao. Genetic diversity of the endangered and medically important Lycium ruthenicum Murr. revealed by sequence-related amplified polymorphism (SRAP) markers. Biochemical Systematics and Ecology. 45, 86-97 (2012).
[2]	H. Cheng, W. Wu, J. Chen, H. Pan, E. Xu, S. Chen, X. Ye, and J. Chen. Establishment of anthocyanin fingerprint in black wolfberry fruit for quality and geographical origin identification. Lwt. 157, 113080 (2022).
[3]	Z. Liu, B. Liu, H. Wen, Y. Tao, and Y. Shao. Phytochemical profiles, nutritional constituents and antioxidant activity of black wolfberry (Lycium ruthenicum Murr.). Industrial Crops and Products. 154, 112692 (2020).
[4]	H. Wang, J. Li, W. Tao, X. Zhang, X. Gao, J. Yong, J. Zhao, L. Zhang, Y. Li, and J.-a. Duan. Lycium ruthenicum studies: Molecular biology, phytochemistry and pharmacology. Food Chemistry. 240, 759-766 (2018).
[5]	R. Di Cagno, R. Coda, M. De Angelis, and M. Gobbetti. Exploitation of vegetables and fruits through lactic acid fermentation. Food microbiology. 33, 1-10 (2013).
[6]	M. Ray, K. Ghosh, S. Singh, and K. C. Mondal. Folk to functional: an explorative overview of rice-based fermented foods and beverages in India. Journal of Ethnic Foods. 3, 5-18 (2016).
[7]	A. Septembre-Malaterre, F. Remize, and P. Poucheret. Fruits and vegetables, as a source of nutritional compounds and phytochemicals: Changes in bioactive compounds during lactic fermentation. Food research international. 104, 86-99 (2018).
[8]	B. N. Abdul Hakim, N. J. Xuan, and S. N. H. Oslan. A comprehensive review of bioactive compounds from lactic acid bacteria: Potential functions as functional food in dietetics and the food industry. Foods. 12, 2850 (2023).
[9]	T. Wang, C. Zhou, Y. Shen, X. Wang, X. Ding, S. Tian, Y. Liu, G. Peng, S. Xue, and J. Zhou. Prevalence of androgenetic alopecia in China: a community‐based study in six cities. British Journal of Dermatology. 162, 843-847 (2010).
[10]	N. Hibberts, A. Howell, and V. Randall. Balding hair follicle dermal papilla cells contain higher levels of androgen receptors than those from non-balding scalp. Journal of Endocrinology. 156, 59-65 (1998).
[11]	F. Lolli, F. Pallotti, A. Rossi, M. C. Fortuna, G. Caro, A. Lenzi, A. Sansone, and F. Lombardo. Androgenetic alopecia: A review. Endocrine. 57, 9-17 (2017).
[12]	L. Wang, S. Mo, G. Zhang, X. Yue, Y. Qu, X. Sun, and K. Wang. Natural phenylethanoid glycoside forsythoside A alleviates androgenetic alopecia by selectively inhibiting TRPV3 channels in mice. European Journal of Pharmacology. 990, 177264 (2025).
[13]	R. Rajput. A scientific hypothesis on the role of nutritional supplements for effective management of hair loss and promoting hair regrowth. J Nutr Health Food Sci. 6, 1-11 (2018).
[14]	A. Kamimura, T. Takahashi, and Y. Watanabe. Investigation of topical application of procyanidin B-2 from apple to identify its potential use as a hair growing agent. Phytomedicine. 7, 529-536 (2000).
[15]	T. Takahashi, T. Kamiya, and Y. Yokoo. Proanthocyanidins from grape seeds promote proliferation of mouse hair follicle cells in vitro and convert hair cycle in vivo. Acta dermato-venereologica. 78, (1998).
[16]	D.-W. Park, H. S. Lee, M.-S. Shim, K. J. Yum, and J. T. Seo. Do kimchi and Cheonggukjang probiotics as a functional food improve androgenetic alopecia? A clinical pilot study. The world journal of men's health. 38, 95 (2020).
[17]	E. Radulescu, A. E. Jaffe, R. E. Straub, Q. Chen, J. H. Shin, T. M. Hyde, J. E. Kleinman, and D. R. Weinberger. Identification and prioritization of gene sets associated with schizophrenia risk by co-expression network analysis in human brain. Molecular psychiatry. 25, 791-804 (2020).
[18]	G. Zhang, J. Xu, Y. Wang, X. Sun, S. Huang, L. Huang, Y. Liu, H. Liu, and J. Sun. Combined transcriptome and metabolome analyses reveal the mechanisms of ultrasonication improvement of brown rice germination. Ultrasonics sonochemistry. 91, 106239 (2022).
[19]	P.-J. Park, B.-S. Moon, S.-H. Lee, S.-N. Kim, A.-R. Kim, H.-J. Kim, W.-S. Park, K.-Y. Choi, E.-G. Cho, and T. R. Lee. Hair growth-promoting effect of Aconiti Ciliare Tuber extract mediated by the activation of Wnt/β-catenin signaling. Life Sciences. 91, 935-943 (2012).
[20]	S. Xiao, Y. Deng, X. Mo, Z. Liu, D. Wang, C. Deng, and Z. Wei. Promotion of hair growth by conditioned medium from extracellular matrix/stromal vascular fraction gel in C57BL/6 mice. Stem cells international. 2020, 9054514 (2020).
[21]	L. Subedi, P. Pandey, J.-H. Shim, K.-T. Kim, S.-S. Cho, K.-T. Koo, B. J. Kim, and J. W. Park. Preparation of topical bimatoprost with enhanced skin infiltration and in vivo hair regrowth efficacy in androgenic alopecia. Drug Delivery. 29, 328-341 (2022).
[22]	H.-C. Wu, X. Fan, C.-H. Hu, Y.-C. Chao, C.-S. Liu, J.-C. Chang, and Y. Sen. Comparison of mitochondrial transplantation by using a stamp-type multineedle injector and platelet-rich plasma therapy for hair aging in naturally aging mice. Biomedicine & Pharmacotherapy. 130, 110520 (2020).
[23]	S. Müller-Röver, K. Foitzik, R. Paus, B. Handjiski, C. van der Veen, S. Eichmüller, I. A. McKay, and K. S. Stenn. A comprehensive guide for the accurate classification of murine hair follicles in distinct hair cycle stages. Journal of investigative dermatology. 117, 3-15 (2001).
[24]	Z.-d. Wang, Y. Feng, L.-y. Ma, X. Li, W.-f. Ding, and X.-m. Chen. Hair growth promoting effect of white wax and policosanol from white wax on the mouse model of testosterone-induced hair loss. Biomedicine & Pharmacotherapy. 89, 438-446 (2017).
[25]	X. Hu, X. Li, S. Wu, X. Jiang, G. Chen, Y. Hu, J. Sun, and W. Bai. Cyanidin-3-O-glucoside and its derivative vitisin A alleviate androgenetic alopecia by exerting anti-androgen effect and inhibiting dermal papilla cell apoptosis. European Journal of Pharmacology. 963, 176237 (2024).
[26]	M. Han, C. Li, C. Zhang, C. Song, Q. Xu, Q. Liu, J. Guo, and Y. Sun. Single-cell transcriptomics reveals the natural product Shi-Bi-Man promotes hair regeneration by activating the FGF pathway in dermal papilla cells. Phytomedicine. 104, 154260 (2022).
[27]	G.-H. Park, K.-y. Park, H.-i. Cho, S.-M. Lee, J. S. Han, C. H. Won, S. E. Chang, M. W. Lee, J. H. Choi, and K. C. Moon. Red ginseng extract promotes the hair growth in cultured human hair follicles. Journal of Medicinal Food. 18, 354-362 (2015).
[28]	M. Kadaja, B. E. Keyes, M. Lin, H. A. Pasolli, M. Genander, L. Polak, N. Stokes, D. Zheng, and E. Fuchs. SOX9: a stem cell transcriptional regulator of secreted niche signaling factors. Genes & development. 28, 328-341 (2014).
[29]	C. Zhang, Y. Li, J. Qin, C. Yu, G. Ma, H. Chen, and X. Xu. TMT-based quantitative proteomic analysis reveals the effect of bone marrow derived mesenchymal stem cell on hair follicle regeneration. Frontiers in pharmacology. 12, 658040 (2021).
[30]	R. Stark, M. Grzelak, and J. Hadfield. RNA sequencing: the teenage years. Nature Reviews Genetics. 20, 631-656 (2019).
[31]	Y. Kong, Z.-C. Feng, Y.-L. Zhang, X.-F. Liu, Y. Ma, Z.-M. Zhao, B. Huang, A.-J. Chen, D. Zhang, and F. Thorsen. Identification of immune-related genes contributing to the development of glioblastoma using weighted gene co-expression network analysis. Frontiers in immunology. 11, 1281 (2020).
[32]	Y. Chen, Z. Fan, X. Wang, M. Mo, S. B. Zeng, R.-H. Xu, X. Wang, and Y. Wu. PI3K/Akt signaling pathway is essential for de novo hair follicle regeneration. Stem Cell Research & Therapy. 11, 1-10 (2020).
[33]	Y. Li, J. Wu, H. Yu, X. Lu, and Y. Ni. Formononetin ameliorates cisplatin-induced hair cell death via activation of the PI3K/AKT-Nrf2 signaling pathway. Heliyon. 10, (2024).
[34]	N. Tia, A. K. Singh, P. Pandey, C. S. Azad, P. Chaudhary, and I. S. Gambhir. Role of Forkhead Box O (FOXO) transcription factor in aging and diseases. Gene. 648, 97-105 (2018).
[35]	X. Qin, X. Wang, K. Xu, X. Yang, Q. Wang, C. Liu, X. Wang, X. Guo, J. Sun, and L. Li. Synergistic antitumor effects of polysaccharides and anthocyanins from Lycium ruthenicum Murr. on human colorectal carcinoma LoVo cells and the molecular mechanism. Food Science & Nutrition. 10, 2956-2968 (2022).
[36]	S. Mishra, A. Tripathi, B. P. Chaudhari, P. D. Dwivedi, H. P. Pandey, and M. Das. Deoxynivalenol induced mouse skin cell proliferation and inflammation via MAPK pathway. Toxicology and applied pharmacology. 279, 186-197 (2014).
[37]	Y. J. Nam, E. Y. Lee, E. J. Choi, S. Kang, J. Kim, Y. S. Choi, D. H. Kim, J. H. An, I. Han, and S. Lee. CRH receptor antagonists from Pulsatilla chinensis prevent CRH‐induced premature catagen transition in human hair follicles. Journal of Cosmetic Dermatology. 19, 3058-3066 (2020).
[38]	Ö. A. Öztürk, H. Pakula, J. Chmielowiec, J. Qi, S. Stein, L. Lan, Y. Sasaki, K. Rajewsky, and W. Birchmeier. Gab1 and Mapk signaling are essential in the hair cycle and hair follicle stem cell quiescence. Cell reports. 13, 561-572 (2015).
[39]	Q. Chen, Z. Wang, Y. Xiong, X. Zou, and Z. Liu. Comparative study of p38 MAPK signal transduction pathway of peripheral blood mononuclear cells from patients with coal-combustion-type fluorosis with and without high hair selenium levels. International journal of hygiene and environmental health. 213, 381-386 (2010).
[40]	H.-C. Huang, H. Lin, and M.-C. Huang. Lactoferrin promotes hair growth in mice and increases dermal papilla cell proliferation through Erk/Akt and Wnt signaling pathways. Archives of dermatological research. 311, 411-420 (2019).
[41]	V. V. Inamdar, H. Reddy, C. Dangelmaier, J. C. Kostyak, and S. P. Kunapuli. The protein tyrosine phosphatase PTPN7 is a negative regulator of ERK activation and thromboxane generation in platelets. Journal of Biological Chemistry. 294, 12547-12554 (2019).
[42]	J. Zhou, X. Kang, H. An, Y. Lv, and X. Liu. The function and pathogenic mechanism of filamin A. Gene. 784, 145575 (2021).
[43]	J.-I. Kang, H.-S. Yoon, S. M. Kim, J. E. Park, Y. J. Hyun, A. Ko, Y.-S. Ahn, Y. S. Koh, J. W. Hyun, and E.-S. Yoo. Mackerel-derived fermented fish oil promotes hair growth by anagen-stimulating pathways. International journal of molecular sciences. 19, 2770 (2018).
[44]	Y. Liang, Y. Zhou, and P. Shen. NF-kappaB and its regulation on the immune system. Cell Mol Immunol. 1, 343-350 (2004).
[45]	X. Lai, T. Liu, Z. Guo, Y. Wang, J. Xiao, Q. Xia, X. Liu, H. Jiang, and X. Wang. In situ formed fluorescent gold nanoclusters inhibit hair follicle regeneration in oxidative stress microenvironment via suppressing NFκB signal pathway. Chinese Chemical Letters. 36, 109762 (2025).
[46]	T. Ito, R. Kageyama, S. Nakazawa, and T. Honda. Understanding the significance of cytokines and chemokines in the pathogenesis of alopecia areata. Experimental Dermatology. 29, 726-732 (2020).
[47]	K. Krieger, S. E. Millar, N. Mikuda, I. Krahn, J. E. Kloepper, M. Bertolini, C. Scheidereit, R. Paus, and R. Schmidt-Ullrich. NF-κB participates in mouse hair cycle control and plays distinct roles in the various pelage hair follicle types. Journal of investigative dermatology. 138, 256-264 (2018).
[48]	F. He, S. Zhang, Y. Li, X. Chen, Z. Du, C. Shao, and K. Ding. The structure elucidation of novel arabinogalactan LRP1-S2 against pancreatic cancer cells growth in vitro and in vivo. Carbohydrate Polymers. 267, 118172 (2021).