D. Don (known as Chuan-Bei-Mu in Chinese) is a prominent medicinal plant utilized in traditional medicine for chronic respiratory ailments. It has garnered global acknowledgment because of its incorporation in many herbal preparations, resulting in a significant increase in demand and, consequently, leading to the decimation of wild populations. The study aimed to obtain regenerated plantlets of . using in vitro propagation techniques and evaluate the accumulation of active metabolites and anti-inflammatory properties from in vitro and natural plant bulbs. UHPLC-Q-TOF/MS analysis identified 267 metabolites. Notably, 118 metabolites showed significantly different intensities between the wild bulbs (WBs) and in vitro tissue culture-regenerated bulbs (RBs). Higher edpetiline amounts were obtained from the RBs, and 14 steroid-related metabolites were elevated in RBs. Both RB and WB extracts had comparable anti-inflammatory abilities and significantly inhibited TNF-α-induced epithelial cell TSLP release. Subsequent mechanistic studies revealed that the efficacy of WB and RB extracts depended on the regulation of the TRPV1/NFAT pathway. These findings highlight the viability of in vitro regeneration and medicinal part replacement as sustainable alternatives to the existing detrimental overharvesting of wild Chuan-Bei-Mu populations.