Enhanced anticancer activity through bidirectional fermentation of Epimedium brevicornum Maxim. with Morchella esculenta

Epimedium brevicornum Maxim. (EB), a traditional Chinese herbal medicine, exhibits notable anticancer and anti-aging activities attributed primarily to flavonoids such as epimedin C and icariin. However, its effective application is limited by low extraction efficiency and incomplete mechanistic understanding. Bidirectional fermentation is an effective microbial bioconversion strategy to enrich bioactive constituents or generate novel metabolites. Herein, the medicinal fungus Morchella esculenta (ME) was used to conduct bidirectional fermentation of EB. Compared with single-fermentation EB extract, the ME/EB extract had 2.39-fold and 1.44-fold greater epimedin C and icariin content, respectively. The ME/EB extract exhibited significant antiproliferative activity against cancer cells, with IC₅₀ values of 215 μg/mL for human cervical cancer HeLa cells and 235 μg/mL for non-small-cell lung cancer A549 cells. Moreover, the ME/EB extract induced cell-cycle arrest at G1/G0 phase in HeLa cells and S phase in A549 cells, while markedly suppressing cell migration and invasion. The ME/EB extract also down-regulated Bcl-2 protein expression and up-regulated Bax, PI3K, and Akt protein expression, thereby promoting cancer cell apoptosis. Therefore, bidirectional fermentation significantly enhanced EB anticancer activity, potentially through modulation of the PI3K/Akt signalling pathway. Overall, ME-mediated bidirectional fermentation of EB shows promise as a novel strategy for developing anticancer agents. © 2026 Elsevier B.V.