Biological & pharmaceutical bulletin最新文献

Lung cancer metastasis is often associated with poor prognosis for patients. Epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET) are important processes in cancer metastasis and are also associated with multidrug resistance (MDR). P-Glycoprotein (P-gp) is one of the efflux transporters responsible for MDR and is anchored to the cell membrane by a family of proteins called ERM (ezrin, radixin, and moesin). The aim of this study was to elucidate the functional changes in P-gp and their mechanisms by continuously inducing EMT and MET in lung cancer cells. EMT was induced by transfecting HCC827 lung cancer cells with the Snail gene, followed by MET induction via dexamethasone. The results suggested that P-gp membrane expression in lung cancer cells increased due to elevated moesin mRNA expression during EMT, followed by increased ezrin expression during MET. The increase in Msn expression did not change during subsequent MET induction, but that in ezrin expression was thought to be further enhanced by the preceding EMT. Furthermore, P-gp membrane expression increased synergistically with the continuous induction of EMT/MET. This study demonstrated that P-gp expression and activity increase at both the EMT and MET stages in cancer metastasis, suggesting that different mechanisms may be involved at each stage.

Chronic liver disease (CLD) poses a significant global health challenge, and liver fibrosis is a crucial process in the pathogenesis of CLD. However, effective interventions to halt and reverse the progression of liver fibrosis remain elusive. This study investigated the potential of the nicotinamide phosphoribosyltransferase (NAMPT) inhibitor FK866 in treating diethylnitrosamine (DEN)-induced liver fibrosis in mice. We first demonstrated that DEN-induced hepatic fibrosis in mice was accompanied by upregulation of hepatic NAMPT and poly (ADP-ribose) polymerase 1 (PARP1) expression. Administration of FK866 inhibited the increase in alanine aminotransferase and aspartate aminotransferase levels and reversed the histopathological changes associated with DEN-induced liver fibrosis. It also suppressed the elevated expression of fibrotic markers, such as fibronectin, collagen IV, laminin, and α-smooth muscle actin. Further studies revealed that this therapeutic effect was achieved by inhibiting the NAD⁺ level, as well as the protein expression of NAMPT, PARP1, and inflammatory factors, including interleukin-1β (IL-1β), IL-6, tumor necrosis factor-α, and P65. In conclusion, FK866 exhibits therapeutic potential for the treatment of liver fibrosis.

In this study, we aimed to develop a clinically applicable risk score using demographic and clinical data available at hospital admission for decompensated heart failure (HF) to identify patients at high risk of medication non-adherence after discharge. In total, 795 patients with HF were enrolled from a prospective, multicenter cohort study. A logistic regression model identified independent predictors of medication non-adherence leading to hospitalization, and we derived a risk score from the partial regression coefficients of these factors. Hospitalization due to medication non-adherence occurred in 6.4% of patients. Independent predictors included dementia (odds ratio [OR]: 2.79; 95% confidence interval [CI]: 1.38-5.65), male sex (OR: 2.50; 95% CI: 1.30-4.79), current smoking (OR: 2.17; 95% CI: 1.07-4.41), non-assisted living (OR: 2.14; 95% CI: 1.17-3.94), and prior HF hospitalization (OR: 1.91; 95% CI: 1.05-3.46). The predictive model showed good discrimination (area under the receiver operating characteristic curve = 0.704). Patients with higher risk scores (4-6 points) had significantly higher rates of all-cause and cardiovascular mortality over 2 years. In conclusion, this pragmatic, admission-based risk score enables early identification of patients at risk for medication non-adherence and may support targeted interventions to improve long-term HF outcomes. Incorporating this score into routine admission processes may help prioritize adherence support for high-risk patients, guide multidisciplinary care planning, and reduce post-discharge complications in HF management.

Treatments to improve the prognosis of heart failure with preserved ejection fraction (HFpEF) are urgently needed. Tanshinone VI (TanVI) is a compound extracted from the roots of Salvia miltiorrhiza Bunge (Lamiaceae) that exerts inhibitory effects on the development of cardiac remodeling under experimental conditions. However, the therapeutic effects of TanVI on HFpEF remain unclear. The present study aims to investigate the therapeutic effects of TanVI in a mouse model of HFpEF. HFpEF mice were prepared by feeding C57BL/6N mice a high-fat diet and providing water containing N^[ω]-nitro-L-arginine methyl ester hydrochloride (L-NAME) for 15 weeks, and TanVI (3 mg/kg/d) or vehicle was administered intraperitoneally using an osmotic pump for 5 weeks until the end of the experiment. The administration of a high-fat diet and L-NAME resulted in cardiac diastolic dysfunction with cardiac hypertrophy and fibrosis. In contrast, treatment with TanVI markedly attenuated cardiac hypertrophy and fibrosis and prevented cardiac diastolic dysfunction. In HFpEF mouse hearts, the phosphorylation levels of mitogen-activated protein (MAP) kinases such as c-Raf, MEK1/2, and extracellular signal-regulated kinase 1/2 (ERK1/2), which regulate cardiac hypertrophy and fibrosis, were elevated. In contrast, treatment with TanVI reversed the phosphorylation levels of c-Raf, MEK1/2, and ERK1/2. The present study showed that TanVI prevented the development of HFpEF by reducing cardiac remodeling. Furthermore, our findings suggest, at least in part, that TanVI attenuates the development of HFpEF by inhibiting the MAP kinase signaling pathway.

Extracellular vesicles (EVs), including exosomes, mediate intercellular communication by transferring lipids, proteins, and nucleic acids. However, the mechanisms determining selective cargo loading into EVs remain poorly understood. Here, we identify the cardiac glycoside ouabain as a selective inhibitor of CD63 loading into EVs. Using a luciferase-based high-throughput assay with CD9- and CD63-tagged reporter cells, ouabain was found to specifically suppress CD63 loading into EVs without affecting CD9 loading into EVs. Ouabain, Na⁺/K⁺-ATPase inhibitor, did not suppress EV secretion but markedly decreased CD63 incorporation. Other cardiac glycoside with strong Na⁺/K⁺-ATPase inhibitory activity, such as bufalin, exhibited similar effects, whereas weak inhibitors did not. Ouabain induced the internalization of Na⁺/K⁺-ATPase (ATP1A1) with CD63, resulting in the disappearance of CD63 from the plasma membrane. Furthermore, ouabain activated autophagy and promoted the colocalization of CD63 with autophagosomes, thereby selectively inhibiting the loading of CD63 into EVs. These effects required both Na⁺/K⁺-ATPase-dependent endocytosis and autophagy, as rapamycin-induced autophagy alone did not remove surface CD63. Our findings reveal a previously unrecognized mechanism in which cardiac glycoside regulates EV cargo composition by coupling Na⁺/K⁺-ATPase-mediated endocytosis with autophagy. Given that endogenous and therapeutic cardiac glycosides are implicated in cardiovascular and cancer biology, this mechanism may broadly influence EV-mediated intercellular communication and represent a potential target for modulating EV functions.

Disruption of osteoblast differentiation can lead to severe bone diseases, such as osteoporosis and osteosclerosis. Our previous study showed that a reduced 4S/6S ratio promotes osteoblast differentiation, linking this specific chondroitin sulfate (CS) modification to bone pathology. This study investigated the effect of forced elevation of the 4S/6S ratio on differentiation. C4ST-1 was found to exhibit overexpression, increasing the 4S/6S ratio, significantly suppressing osteoblast differentiation, as evidenced by reduced Akp2 gene expression and ALP activity. This inhibition was far more potent than that caused by the enzymatic removal of CS. Notably, treating C4ST-1-overexpressing cells with chondroitinase reduced differentiation inhibition to a level similar to that in mock cells treated with chondroitinase. These results suggest that the strong inhibition was due to the excessive 4-sulfated CS produced by C4ST-1, implying a mechanism distinct from the inhibition caused by a lack of CS. To elucidate this mechanism, we investigated the potential feedback loop. The increase in 4-sulfated CS from C4ST-1 overexpression enhanced Wnt3a expression, which upregulated p53 expression. Pharmacological inhibition of β-catenin and p53 partially restored Akp2 expression, confirming their roles as key mediators. We propose that a high 4S/6S ratio activates a Wnt/β-catenin-p53 axis, where p53 acts as a brake on differentiation. Our findings highlight the critical role of CS sulfation patterns in fine-tuning the osteoblast fate.

Nitric oxide (NO) plays diverse roles in tumor biology, including modulation of blood flow and induction of cell death at high concentrations. In this study, we synthesized a novel NO donor derived from valproic acid (NVA) and investigated its cytotoxic mechanism in human pancreatic cancer cells. NVA released approximately 40% of its total nitrate/nitrite (NO_x) immediately after dissolution in phosphate-buffered saline and then remained almost unchanged for 72 h, indicating a rapid initial NO release followed by stabilization. NVA significantly decreased the viability of BxPC-3 cells, whereas valproic acid (VA) alone had little effect. Flow cytometric analysis using Annexin V revealed that NVA-induced cell death was not inhibited by the pan-caspase inhibitor Z-VAD-FMK. Furthermore, Western blotting showed no cleavage of caspase-3 or poly(ADP-ribose) polymerase (PARP) following NVA exposure, suggesting that apoptosis was not the major pathway. These findings indicate that NVA induces NO-dependent, caspase-independent cell death, distinct from classical apoptosis. The present study provides fundamental insights into the potential use of VA-based NO donors as antitumor agents against pancreatic cancer.

A combination of anti-programmed death-1 (anti-PD-1) immunotherapy and radiation therapy (RT) has shown good clinical outcomes. Previously, we had reported that enhanced anticancer immune response can relieve the hypoxia in tumors after treatment with anti-PD-1 agents. In this study, we investigated the relationship between tumor hypoxia and anticancer immunity in the combination therapy. An anti-PD-1 antibody was administered to mice with CT26 tumor on days 0 and 3. Tumors were irradiated with X-rays on days 0, 1, 2, and 3. Hypoxia in tumors was non-invasively examined with a hypoxia positron emission tomography (PET) probe, [¹⁸F]fluoromisonidazole ([¹⁸F]FMISO). The percentages of immune and hypoxic cells were analyzed using flow cytometry with pimonidazole. One day after four cycles of irradiation (day 4), no change was observed in the numbers of CD8⁺ T cells and hypoxic cells within the tumor in the combination therapy group. On day 10, a later timepoint, the numbers of CD8⁺ T cells was increased. The number of hypoxic cells was decreased, but [¹⁸F]FMISO accumulation in the tumors was unchanged. Previous studies had shown that CD8⁺ T cells remodel tumor vasculature in anti-PD-1 therapy, leading to the reduction in hypoxia. Our results also suggest that the recruited CD8⁺ T cells reduced hypoxia in tumors irradiated with X-rays, although [¹⁸F]FMISO-PET did not indicate these changes. In conclusion, the combination therapy with anti-PD-1 antibody and RT reduced the hypoxic levels in tumors in association with the increased infiltration of CD8⁺ T cells.

Antimicrobial resistance (AMR) has become a major global health threat, prompting the implementation of antimicrobial stewardship (AS) programs to optimize antimicrobial use. Pharmacists play a vital role in these efforts. In Japan, a series of national policy measures has been launched as part of the 2016 National Action Plan on AMR, and systems have been established in parallel to support the active involvement of pharmacists with advanced expertise in infectious diseases. This narrative review provides an overview of the expanding contributions of pharmacists to AS across diverse healthcare settings in Japan. In large hospitals with relatively abundant human resources, pharmacists have promoted post-prescription review, dose optimization, and de-escalation, resulting in reduced use of broad-spectrum antibiotics, shortened treatment durations, cost savings, and even lower resistance rates. Further, in small- and medium-sized hospitals, where infectious disease specialists are often unavailable, pharmacist-led stewardship efforts are gradually emerging. Community pharmacists have also expanded AS efforts outside hospital settings by surveilling outpatient antibiotic prescriptions and identifying leftover antibiotics during home visits. In addition, multicenter collaborative studies and analyses using large-scale administrative claims databases are contributing to the generation of robust real-world evidence, facilitating sustained surveillance and policy evaluation. To maintain and further enhance these efforts, fostering the next generation of AS professionals and developing novel indicators and predictive models are essential. Pharmacist-led AS in Japan is expected to continue progressing, with sustainable contributions needed across healthcare situations of all field.