Caesalpinia sappan L. has exhibited various pharmacological effects, yet its anticancer activities against colorectal cancer (CRC) and underlying molecular mechanisms remain unclear. This study investigated the anticancer properties of an ethanol extract of C. sappan L. (CSE) against CRC cells, focusing on the identification of bioactive compounds and their mechanisms of action. A network pharmacology analysis was conducted to identify potential CRC targets and bioactive compounds of CSE, using LC-MS for compound identification. The anticancer effects of CSE were then validated through in vitro and in vivo models of CRC. The network pharmacological approach identified 87 overlapping genes between CSE targets and CRC-related genes, with protein–protein interaction analysis highlighting 33 key target genes. CSE inhibited cell proliferation in human CRC cell lines, including HCT 116, KM12SM, HT-29, and COLO 205, and induced apoptosis via caspase 3/7 activation. Western blot analyses confirmed the modulation of critical signaling pathways, including STAT3, AKT, and mitogen-activated protein kinases. Furthermore, CSE significantly suppressed tumor growth in MC38 CRC-bearing mice. These findings suggest that CSE possesses substantial potential as a natural anticancer agent for CRC treatment, highlighting the need for further exploration in therapeutic development.

Bojungikki-Tang (BJIKT) is traditionally used to enhance digestive function and immunity. It has gained attention as a supplement to chemotherapy or targeted therapy owing to its immune-boosting properties. This study aimed to evaluate the synergistic anti-tumor effects of BJIKT in combination with pembrolizumab in a preclinical model. MHC I/II double knockout NSG mice were humanized with peripheral blood mononuclear cells (PBMCs) and injected subcutaneously with H460 lung tumor cells to establish a humanized tumor model. Both agents were administered to evaluate their impact on tumor growth and immune cell behavior. Immunohistochemistry showed decreased exhaustion markers in CD8(+) and CD4(+) T cells within the tumor, indicating enhanced T cell activity. Additionally, RNA sequencing, transcriptome analysis, and quantitative PCR analysis were performed on tumor tissues to investigate the molecular mechanisms underlying the observed effects. The results confirmed that BJIKT improved T cell function and tumor necrosis factor signaling while suppressing transforming growth factor-β signaling. This modulation led to cell cycle arrest and apoptosis. These findings demonstrate that BJIKT, when combined with pembrolizumab, produces significant anti-tumor effects by altering immune pathways and enhancing the anti-tumor immune response. This study provides valuable insights into the role of BJIKT in the tumor microenvironment and its potential to improve therapeutic outcomes.

Background: In the realm of cardiovascular and renal health, the modulation of oxidative stress and inflammation within the cardio-renal axis plays a pivotal role. This study investigates the therapeutic potential of Gunryeong-tang (GRT), a traditional Korean herbal formula combining Oryeongsan (Wulingsan) and Sagunza-tang (Sijunzi-tang), specifically in addressing cardio-renal axis dysfunction induced by pulmonary arterial hypertension (PAH-r). Methods: PAH-r models were induced through intraperitoneal injection of monocrotaline (MCT) at a daily dose of 50 mg/kg for 5 days to simulate pulmonary hypertension, a common cause of cardio-renal syndrome. The primary focus is on evaluating its anti-fibroinflammatory effects and its ability to modulate oxidative stress and inflammation. The treatment group received oral administration of GRT (100 mg/kg/day). Results: Various parameters were assessed, including right ventricular (RV) enlargement and fibrosis, lung weight, pulmonary fibrosis, expression of cardiac hypertrophy markers, and involvement of specific pathways (TGF-β/p-Smad2 and HMGB-1/TLR4/MyD88/NF-κB). Renal dysfunction markers, such as blood urea nitrogen (BUN), creatinine clearance rate (Ccr), glomerular dilatation, tubular fibrosis, and protein expression of kidney injury molecule-1 (KIM-1) and neutrophil gelatinase-associated lipocalin (NGAL), were evaluated to highlight the formula's anti-fibroinflammatory effects. Treatment resulted in significant improvements in the PAH-r group. It alleviated RV enlargement and fibrosis, suppressed lung weight increase, and mitigated pulmonary fibrosis. The administration downregulated cardiac hypertrophy markers (CTGF, TGF-β, and α-SMA mRNA levels) and inhibited the TGF-β/p-Smad2 and HMGB-1/TLR4/MyD88/NF-κB pathways, showcasing GRT's anti-fibroinflammatory properties. Additionally, the treatment ameliorated renal dysfunction, as evidenced by reduced BUN and Ccr levels, improved glomerular dilatation, and reduced protein expression of KIM-1 and NGAL. GRT demonstrates significant therapeutic potential in managing cardio-renal axis dysfunction induced by PAH-r, emphasizing its multifaceted benefits in modulating oxidative stress and inflammation within this axis. Conclusions: This study provides valuable insights into the underlying mechanisms, highlighting the modulation of cardiac hypertrophy markers and specific pathways. These findings underscore the promising role of GRT as a viable treatment option for cardio-renal axis dysfunction associated with PAH-r.

Immunogenic cell death (ICD) converts dying tumor cells into an anticancer vaccine, yet whether protection depends on how those cells are prepared and when they are given has rarely been examined. Using mitoxantrone (MTX), a clinically approved small molecule classified as an ICD inducer, we examined both variables in the murine CT-26 colon carcinoma model. MTX reduced viability in a concentration-dependent manner and induced extracellular ATP release. The total extent of cell death, however, was similar across concentrations, indicating that the concentration-dependent loss of viability was not accompanied by a proportional increase in cell death. What differed was the composition of the dying population, which shifted from early to late apoptotic cells as the concentration increased. Prophylactic vaccination with MTX-treated cells protected BALB/c mice, leaving 75.0–87.5% of animals tumor-free after vaccination at 1 μM and 37.5–62.5% at 3 μM across independent experiments. Protection increased with the interval between vaccination and challenge, with 0%, 50.0%, and 87.5% of animals tumor-free after simultaneous injection, a 3-day interval, and a 7-day interval, respectively. Cells killed completely by freeze–thaw were inactive as a vaccine, and cisplatin-treated cells conferred only a marginal delay, whereas responders to MTX-treated cells resisted rechallenge for the remainder of the observation period. Taken together, these results show that use of an ICD-inducing drug alone is not sufficient for effective dying-cell vaccination and that protective efficacy is influenced by vaccine preparation and by the interval between vaccination and tumor challenge, rather than by the extent of cell death alone.