Excessive oxidative stress and inflammatory responses are associated with the development of various diseases, including cancer. Glucosinolates (GSLs) are phytochemicals known for their antioxidant properties, and doubled haploid lines (DHLs) of Brassica rapa with high GSL contents (HGSL) were intentionally developed from two edible subspecies of Brassica rapa: B. rapa subsp. trilocularis and B. rapa subsp. chinensis. The purpose of the present study is to assess the capacity of HGSL DHLs to mitigate oxidative stress and inflammation in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, compared to pak choi as a parental control. Our findings demonstrate that HGSL DH lines effectively suppressed the expression of inducible nitric oxide synthase, leading to the reduced levels of nitric oxide at non-toxic concentrations. Additionally, these lines exhibited scavenging activity against reactive oxygen species and free radicals. The enhanced antioxidant capacity of HGSL DHLs was mechanistically attributed to the upregulation of antioxidant enzymes, such as NADPH quinone oxidoreductase 1 (NQO1), the glutamate–cysteine ligase catalytic subunit (GCLC), and heme oxygenase-1 (HMOX1). Furthermore, we confirmed that these effects were mediated through the nuclear factor erythroid 2-related factor 2 (NRF2) signaling pathway via p38 phosphorylation. Moreover, HGSL DHLs demonstrated inhibitory effects on pro-inflammatory cytokines and signal transducers and activators of transcription 3 (STAT3) phosphorylation. Collectively, our results indicate that HGSL DHLs possess better antioxidant and anti-inflammatory properties compared to the parental control pak choi in LPS-stimulated RAW264.7 cells, suggesting that HGSL DHLs of Brassica rapa could be considered as a beneficial daily vegetable for reducing the risk of inflammation-associated diseases.

Chemoprevention is a method of health control in modern industrialized societies. Traditional breeding (hybridization) has been widely used to produce new (sub)species with beneficial phenotypes. Previously, we produced a number of doubled haploid (DH) lines of Brassica rapa with a high glucosinolate (GSL) content. In this study, we evaluated the anticancer activities of extracts from three selected high-GSL (HGSL)-containing DH lines (DHLs) of Brassica rapa in human colorectal cancer (CRC) cells. The three HGSL DHL extracts showed anti-proliferative activities in the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium assay and pro-apoptotic activities in the cell cycle or annexin V analysis with the induction of pro-apoptotic protein expression in CRC cells. Mechanistically, HGSL DHL extracts inhibited the NF-κB and ERK pathways, leading to a reduction in the nuclear localization of NF-κB p65. In addition, reactive oxygen species were induced by HGSL DHL extract treatment in CRC cells. In conclusion, our data suggest that the newly developed HGSL DHLs possess enhanced anticancer activities and are potentially helpful as a daily vegetable supplement with chemopreventive activities.

PSY is a three-herb formula comprising Patriniae Radix, Mori Cortex Radicis, and Coix Seed that was previously shown to inhibit STAT3 signaling in colorectal cancer (CRC) cells. Here, we investigated molecular and metabolic responses to PSY, its antitumor activity in an HCT116 xenograft model, and its interactions with chemotherapy in vitro. Representative chemical markers were qualitatively verified by UPLC-ESI-QTOF MS/MS. Exploratory RNA sequencing with two biological replicates per group highlighted stress-associated and metabolic processes for further investigation. PSY increased BiP, phosphorylated eIF2α, ATF4, and CHOP levels, intracellular reactive oxygen species (ROS) levels, and mitochondrial membrane depolarization, while reducing the phosphorylation of PI3K/AKT/mTOR- and ERK-associated proteins. PSY increased the p-AMPK/AMPK ratio and decreased the p-ACC/ACC ratio. PSY reduced several lipid-biosynthetic transcripts and decreased the relative abundance of selected fatty acids in xenograft tumors. Oral administration of PSY suppressed HCT116 xenograft growth without significant body-weight loss. Combination-index (CI) analysis indicated CI-defined in vitro synergistic interactions with 5-fluorouracil (5-FU) at multiple concentration pairs, with interaction patterns varying across HCT116, HT-29, and SW480 cells. Direct cell counting supported the combination response in HCT116 and SW480 cells. The combination increased apoptosis and reduced STAT3 and NF-κB p65 phosphorylation relative to single treatments. These findings characterize concurrent, partly discordant signaling and metabolic responses to PSY and support further preclinical evaluation of its in vitro interaction with 5-FU.