Haixia Li¹, Xiaolan Chen², Guangjie Zhang², Li Du², Yuxin Lu², Xiaochen Cheng², Shuchen Liu², Min Li², Qinglin Zhang^2,, Fengjun Xiao^2,

Orychophragmus violaceus (OV), also known as the February orchid, has been used in traditional Chinese medicine for thousands of years. It possesses various physiological activities. In this study, the major components of OV seed alkaloid extract (OVS-2) were identified through liquid chromatography-mass spectrometry (LC-MS). The protective effect of OVS-2 against radiation-induced oxidative stress in rat intestinal crypt epithelial IEC-6 cells was determined. C57 mice underwent abdominal irradiation with ⁶⁰Co gamma rays to induce radiation-induced intestinal injury (RIII), followed by OVS-2 treatment. The efficacy of OVS-2 against RIII was evaluated based on changes in survival rate, body weight, histopathological staining, and intestinal biomarkers. Serum inflammatory cytokine levels and mRNA expression levels in the tissues were also measured. Potential mechanisms were explored using transcriptomic sequencing. LC-MS analysis revealed that the top 20 monomeric components of OVS-2 were primarily alkaloids. We demonstrated that the expression levels of malondialdehyde (MDA) and lipid peroxidation markers in OVS-2-treated IEC6 cells were significantly reduced under radiation-induced oxidative damage, while maintaining mitochondrial integrity. OVS-2 treatment improved the survival rate of irradiated mice, as evidenced by increased villus length and crypt depth in the histopathological sections. Increased expressions of intestinal biomarkers Ki67, Muc2, and lysozyme were observed after OVS-2 treatment, along with decreased expressions of inflammatory cytokines IL-6, IL-1β, and TNF-α in serum and intestinal tissues. Transcriptomic sequencing of intestinal tissues revealed the enrichment of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways that were mainly related to immune regulation. Results suggest that the active components of OV mainly consist of alkaloids that exhibit antioxidant, anti-inflammatory, and intestinal mechanical barrier-protective effects.

February orchids, RIII, alkaloid monomer, oxidative stress, transcriptomic sequencing