Anticancer Activity of Mineral-Supplemented Organically Cultivated Carrot on HT-29 Cells and Its Anti-Inflammatory Effect on Mice Splenocytes

Abstract

Carrot (Daucus carota) is one of the world’s most significant root vegetables, with various bioactivities. This study aimed to investigate the anticancer activity and anti-inflammatory effects of natural dream cultivation carrot (NC). Natural dream cultivation is a cultivation method based on organic farming incorporating minerals. An MTT assay was used to evaluate the inhibitory rate of carrot samples on HT-29 human colon cancer cells, and qPCR was used to assess the mRNA expression of the cell cycle and apoptosis-related genes in the cancer cells. The nitrite oxide (NO) concentration was determined using the Griess method. The levels of inflammatory cytokines in LPS-induced mouse splenocytes were determined using an enzyme-linked immunosorbent assay, and the activity of NK cells was determined using LDH analysis. The results revealed that NC effectively inhibited cancer cell growth rate. Moreover, NC upregulated the mRNA expression of cell-cycle-arrest-related genes (p53 and p21) and apoptosis-related genes (Bim, Bad, Bax, Bak, caspase-9, and caspase-3) in cancer cells while downregulating the expression of anti-apoptotic genes, Bcl-2 and Bcl-xL. NC inhibited NO production and the release of inflammatory cytokines (TNF-α, IL-6, IL-1β, IFN-γ, and IL-12) in LPS-induced mouse splenocytes. NC also demonstrated the ability to stimulate NK cell activation. This study explored the potential mechanisms underlying carrots’ anticancer and anti-inflammatory properties by investigating their inhibitory effects on cancer cells and regulating the inflammatory response. The innovative mineral-supplemented organic cultivation method, as explored in this study, opens new avenues for harnessing the potential of carrots as a functional food source with promising applications in cancer and inflammation management. This research not only provides insights into the bioactive potential of carrots but also contributes to the future development of novel dietary interventions and therapeutics.

Anti-Inflammatory Effects and Immunomodulatory Efficacy of Unitein (Fermented Glycine max, Panax ginseng, and Chenpi Mixture) in RAW 264.7 Macrophages and Mice Splenocytes along with Component Analysis

“…Although moderate NO production assists in pathogen clearance, excessive NO production might induce tissue damage and exacerbate infammatory reactions [26]. Research fndings suggest that under immune stimulation, such as exposure to LPS, the expression and activation of iNOS in RAW 264.7 cells and mice splenocytes increase, resulting in elevated NO production that participates in immune and infammatory regulation [4,5]. Our evaluation of NO generation after LPS treatment revealed signifcantly lower NO production in the sample-treated groups than in the LPS-treated group.…”

“…During infection, trauma, and tissue damage, IL-6 production increases, promoting immune cell activation and infammatory responses [32]. Based on multiple research fndings, it has been shown that LPS has an impact on RAW 264.7 cells and splenocytes by increasing the levels of infammatory cytokines, including TNF-α, IL-1β and IL-6 [4,5,[33][34][35].…”

“…Te immune system safeguards the body against infections by identifying and eliminating pathogens [3]. RAW 264.7 cells and splenocytes are cellular models commonly utilized for investigating the cellular and molecular mechanisms associated with immunity and infammation [4,5]. RAW 264.7 cells are a widely employed murine macrophage cell line, are derived from peritoneal macrophages of BALB/ c mice, and are favored for their ease of cultivation and maintenance of stable immunological characteristics, which has been extensively utilized to explore immune and infammatory responses [6].…”