In 2007, the Japanese Orthopaedic Association proposed the concept of locomotive syndrome, a comprehensive description of conditions involving the functional decline of the locomotor system. Locomotive syndrome includes bone-related diseases such as osteoporosis, joint cartilage and disc-related diseases such as osteoarthritis and lumbar spondylosis, and sarcopenia and locomotive syndrome-related diseases. If left untreated, these diseases are likely to reduce mobility, necessitating nursing care. To prevent the progression of locomotive syndrome, a daily exercise routine and well-balanced diet are important, in addition to recognizing one’s own decline in mobility. Therefore, research on the effectiveness of functional ingredients in the prevention and suppression of locomotive syndrome progression is ongoing. In this review, we summarize the latest reports on the effectiveness of five functional ingredients, namely, epigallocatechin gallate, resveratrol, curcumin, ellagic acid, and carnosic acid, in the treatment of osteoarthritis, osteoporosis, and rheumatoid arthritis, which are considered representative diseases of the locomotive syndrome.

Statins are cholesterol-lowering drugs often used for the treatment of dyslipidemia. Statins also exert anti-cancer effects by inhibiting hydroxymethylglutaryl-CoA reductase (HMGCR), a rate-limiting enzyme in cholesterol synthesis. We previously reported that the susceptibility to statin treatment differs among cancer cells and that functional E-cadherin expression on the plasma membrane could be a biomarker of statin sensitivity in cancer cells. However, the detailed qualitative and molecular differences between statin-sensitive and statin-resistant cancer cells remain unclear. Here, we explored novel parameters related to statin sensitivity by comparing gene expression profiles and metabolite contents between statin-sensitive and statin-resistant lung cancer cell lines. We found that the expression of most cholesterol synthesis genes was lower in the statin-sensitive cancer cell line, HOP-92, than in the statin-resistant cancer cell line, NCI-H322M. Moreover, HOP-92 cells originally exhibited lower levels of CoA and HMG-CoA. Additionally, atorvastatin decreased the mRNA expression of PANK2, a rate-limiting enzyme in CoA synthesis. Atorvastatin also reduced the mRNA levels of the cholesterol esterification enzyme SOAT1, which was consistent with a decrease in the ratio of cholesterol ester to total cholesterol in HOP-92 cells. Our data suggest that the cholesterol synthetic flow and CoA content may be limited in statin-sensitive cancer cells. We also suggest that CoA synthesis and cholesterol storage may fluctuate with atorvastatin treatment in statin-sensitive cancer cells.

Using 2-pyrazole oxazolidine-4-one derivative 1 as the lead compound, a series of novel 2-heteroaryl-oxazolidine-4-one derivatives were designed and synthesized by replacing pyrazole moiety with other heterocycles, including methyl pyrazole, oxazole, thiazole, triazole, and reverse pyrazoles, based on the principle of bioisosterism. The structures of target compounds were established by ¹H-NMR, ¹³C-NMR, and electrospray ionization (ESI)-MS. Majority of these compounds showed moderate to good inhibitory activities against hepatitis B virus (HBV) DNA proliferation with low cytotoxicities. Especially, compound 7g showed the most potent anti-HBV activity with EC₅₀ value of 0.059 µM, accompanied with better selectivity index (SI) value than that of lead compound. Our research revealed the structure–activity relationship (SAR) of these newly designed compounds and compound 7g was identified as the potential HBV capsid assembly modulator.

Sudachitin, a polymethoxyflavone found in sudachi peel, has been reported to improve hyperlipidemia in humans, and is thus attracting research attention. However, its effect on cardiac function remains unclear. We investigated the mechanisms underlying the chronotropic and inotropic effects of sudachitin on rat atria. Sudachitin (0.3–30 µM) produced concentration-dependent positive chronotropic and inotropic effects. Other polymethoxyflavones, including demethoxysudachitin (0.3–30 µM) and nobiletin (0.3–30 µM), also produced positive chronotropic and inotropic effects; however, the maximum efficacy of all polymethoxyflavones, including sudachitin, was lower than that of isoproterenol. Propranolol (0.1 µM) did not affect the positive chronotropic and inotropic effects of sudachitin. The concentration–response curves for the chronotropic and inotropic effects of dibutyryl-cAMP (1–100 µM) were shifted to the left upon pretreatment with sudachitin (3, 10 µM). Phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine 1 µM or milrinone 10 µM) alone, sudachitin alone (10, 30 µM), and a combination of phosphodiesterase inhibitors and sudachitin exhibited positive chronotropic and inotropic effects, whereas the lack of any interaction between each phosphodiesterase inhibitor and sudachitin indicated an additive effect of the two substances. These results suggest that sudachitin-induced positive chronotropic and inotropic effects similar to those of other polymethoxyflavones, but its maximum efficacy was lower than that of isoproterenol. Both demethoxysudachitin and nobiletin exhibited similar positive chronotropic and inotropic effects, indicating that these effects are not specific to sudachitin, but are common to polymethoxyflavones. The mechanism of action of sudachitin was associated with the enhancement of cAMP-dependent pathways, without the involvement of β-adrenoceptors.

Effective blood sampling times, beyond trough and peak levels, have not been determined for estimating vancomycin’s area under the concentration–time curve (AUC) using the Bayesian software. The aim of this study was to evaluate the accuracy of AUC estimation at different blood sampling times during the same dosing interval at steady state utilizing data from a prior phase I trial of vancomycin. Six healthy adult participants were sampled following intravenous administration of 1 g vancomycin for 1.5 h every 12 h. The AUC was estimated using four software packages and four population pharmacokinetic models. Accuracy was assessed using bias (difference between the estimated and reference AUC) and imprecision (absolute percentage difference between the estimated and reference AUC). The accuracy varied with the sampling time. The optimal two-point sampling times were determined to be 2.5 and 5.5 h post-injection using software packages for EasyTDM, Practical AUC-guided therapeutic drug monitoring (TDM), and Anti-MRSA Agents TDM Analysis Software (incorporating Rodvold, Yamamoto, and Yasuhara models). In these estimations, the mean bias (range, −1.7 to 9.5 µg·h/mL) was unbiased and the mean imprecision (range, −3.0% to 5.0%) was precise. The optimal one-point sampling time was 5.5 h post-injection for Anti-MRSA Agents TDM Analysis Software, which incorporated the Yamamoto and Yasuhara models. In conclusion, optimal blood sampling times may vary depending on the software and model used. Our findings suggest that identifying specific sampling times could improve the efficacy of TDM in clinical practice.

We have developed a novel cross-immune antigen vaccine platform for influenza A virus. The vaccine antigen is a fusion protein of headless hemagglutinin (HA) and matrix protein 1 (M1), which possess B-cell and T-cell epitopes, respectively, that are conserved among subgroup A viruses. The single molecule of headless HA and M1 fusion protein forms an oligomer by self-assembly of M1. T-Helper 1 (Th1) and Th2 cells were activated in an antigen-specific manner in lymphocyte cultures of antigen-immunized mice. The antigen-immunized antiserum neutralized influenza virus A subtypes H1N1 and H3N2, and intranasal administration of the antigen reduced mortality to less than 30% in a protection assay.

Although medication oversupply results in waste of medications, triggers of medication oversupply remain unclear. This nationwide retrospective cohort study aimed to identify associated factors and causes of chronic disease medication oversupply in Japan by quantitative and qualitative analyses. Data of financial year 2019 from a large insurance claims database were used. Excess days, which represent medication oversupply in days, were calculated for each of the major five classes of chronic disease medications. For each class, the two cohorts were formed, consisting of individuals aged ≥55 years with either excessive oversupply or normal supply according to excess days. Oversupply-associated factors were subjected to quantitative analyses using logistic regression models, which included excessive oversupply vs. normal supply as an outcome variable. A qualitative analysis to identify causes of oversupply was performed by reviewing the medication history of 50 individuals randomly selected from each excessive oversupply cohort, and causes were classified into seven categories. Oversupply-associated factors in all classes were greater frequency of early supply (≥6 vs. <3 times/10 supplies, odds ratio (OR) 5–9 for all classes), inpatient prescription (included vs. not included, OR 3–5), and higher number of concomitant ingredients (≥16 vs. 1–5 ingredients, OR 3–5). The most common category of causes for oversupply in all classes was the “early supply of medications prescribed by a single facility.” The factors and causes of oversupply might reflect the unique features, rules, and customs of Japan’s healthcare system. This finding might help in developing measures for reducing medication oversupply.

Royal jelly (RJ), a natural product secreted by honeybees, is widely used in topical skincare products to help maintain cutaneous homeostasis. Despite its popularity, the mechanism through which RJ exerts its effects on the skin has not been fully elucidated. This study aimed to explore the impact of RJ on the proliferative ability and senescence of human primary epidermal keratinocytes (HPEKs). Our data suggested that epidermal equivalents became thicker with more p63-expressing proliferative cells upon RJ addition to the culture medium. In a two-dimensional culture system, we evaluated the effect of RJ on the proliferation of HPEKs and observed only a slight increase in cell proliferation. This suggests that RJ does not significantly enhance the proliferation of HPEKs in the short term. However, long-term culture experiments demonstrated enhanced population doubling in the RJ-treated group, indicating that RJ inhibits senescence. RJ was found to suppress cellular senescence by modulating the expression levels of ΔNp63, p16, and p21. These results were further supported by the identification of major fatty acids, such as 10-hydroxy-2-decenoic acid, in RJ. Our findings indicate that RJ can maintain epidermal stem cell properties by repressing cellular senescence, providing insights into its mechanism of action in skincare applications.

Retinoid-related orphan receptor-γ (RORγ) is a nuclear receptor that plays important roles in the development and activation of T helper type-17 (Th17) cells. In this study, we characterized the pharmacological profile of JTE-151 ((4S)-6-[(2-chloro-4-methylphenyl)amino]-4-{4-cyclopropyl-5-[cis-3-(2,2-dimethylpropyl)cyclobutyl]isoxazol-3yl}-6-oxohexanoic acid), which is a novel RORγ antagonist identified by our group. JTE-151 dissociated co-activator peptide from the human RORγ-ligand binding domain (LBD) and recruited co-repressor peptide into human RORγ-LBD, and potently inhibited the transcriptional activity of RORγ of human, mouse and rat. JTE-151 also demonstrated high selectivity against other receptors in nuclear receptor family. JTE-151 suppressed the differentiation of mouse naïve CD4⁺ T cells into Th17 cells without affecting the differentiation of those cells into other CD4⁺ T cell subsets in vitro. In addition, JTE-151 inhibited the production of interleukin-17 (IL-17) but not interferon-γ (IFN-γ) and IL-4 from activated human helper T cells in vitro. Furthermore, treatment with JTE-151 suppressed the production of IL-17 in antigen-sensitized mice and ameliorated the severity of arthritis in mice with collagen-induced arthritis regardless of treatment start date. Based on these results, we reasoned that JTE-151 could serve as a novel therapeutic compound for various autoimmune diseases linked to Th17 cells, such as psoriasis and rheumatoid arthritis.

Cutaneous colonization with Staphylococcus aureus (SA) is frequently observed in patients with atopic dermatitis. SA produces a wide variety of bacterial toxins, among which δ-toxin was found to induce degranulation of mast cells. Degranulation of mast cells could enhance bacterial clearance and protection from future SA infection but lead to exacerbation of atopic dermatitis. Because it remains to be determined how δ-toxin triggers degranulation, we investigated δ-toxin-induced changes in murine bone marrow-derived cultured mast cells in this study. We found that δ-toxin-induced degranulation could be classified into two phases, an early Ca²⁺-independent and a late Ca²⁺-dependent phase. Recent studies suggest that NOD-like receptor family, pyrin domain containing 3 is involved in the degranulation of mast cells, raising a possibility that leakage of K⁺ induced by δ-toxin is involved in the Ca²⁺-independent phase. However, Ca²⁺-independent degranulation remains unchanged although Ca²⁺-influx and degranulation induced by δ-toxin were significantly suppressed in the presence of high concentrations of K⁺. Because actin depolymerization was reported to induce degranulation in the absence of Ca²⁺ in the permeabilized rat peritoneal mast cells, a slow but steady decrease in the amount of filamentous actin observed here may be involved in Ca²⁺-independent degranulation induced by δ-toxin. Although Mas-related G protein-coupled receptor (MRGPR) X2 in humans and Mrgprb2 in mice are regarded as the receptors responsible for immunoglobulin E-independent degranulation, δ-toxin-induced degranulation remained unchanged in Mrgprb2^−/− mast cells. Our findings pave the way for identification of the target receptors of δ-toxin.

Cephaibol A was isolated from a freshwater fungus Acremonium tubakii BMC-58 extract which composed of 16 amino acids and featuring multiple α-aminoisobutyric acid. We investigated the cytotoxicity of cephaibol A on MDA-MB-231 cells to elucidate its potential antitumor activity and mechanism. The study found that cephaibol A concentration-dependently blocked the cell cycle in S phase and inhibited cell proliferation. Meanwhile, cephaibol A could reduce the migration and invasion abilities of MDA-MB-231 cells. Further studies proved that cephaibol A caused mitochondrial dysfunction and increased reactive oxygen species (ROS) accumulation. Mitochondrial membrane potential (ΔΨm) assay suggested that cephaibol A induced apoptosis by affecting Bcl-2, Bax and cytochrome c levels, thus decreasing ΔΨm and activating the caspase cascade reaction. Moreover, cephaibol A significantly inhibited tumor growth and improved survival rates in the MDA-MB-231 cell mice model. These findings established cephaibol A as a potential antitumor agent that inhibited tumor cell proliferation in vitro and in vivo by affecting mitochondrial dysfunction and inducing apoptosis in MDA-MB-231 cells through structural damage.

Anacardic acid (AA) was first detected in the shells of cashew nuts, Anacardium occidentale, and is known to possess inhibitory activity against acetyltransferases. Recently, several anacardic acid derivatives (AAds) were isolated from the wild fungus, Tyromyces fissilis, which has been reported as xanthine oxidase inhibitors. In the present study, we investigated whether nine AAds function as acetyltransferase inhibitors. Screening analyses were performed by incubating the enzyme protein (P300/CBP-associated factor; PCAF) and the substrate protein (histone H1) with radioisotope-marked acetyl-CoA, showing that two of the nine derivatives, namely, AAd7 and AAd11, inhibited the acetyltransferase activity of PCAF at concentrations of 50 and 100 µM, respectively. The inhibition intensities were similar to those of the original compound, AA, and the inhibitory effects of these derivatives increased in a concentration-dependent manner. Docking simulations suggested the possibility that AA, AAd7, and AAd11 might bind the same active pocket of PCAF. These results suggest that the AAds can be used as acetyltransferase inhibitors. In contrast, there were no significant differences in the molecular structure of AA and its derivatives; however, these small differences in the functional groups on the alkyl side chain on salicylic acid reduced the acetyltransferase inhibitor activity or newly produced proteolytic activity.

In this study, we attempted to enhance the delivery of minoxidil (MXD) nanocrystals into hair follicles for efficacious hair growth treatment. We applied a bead milling method and designed an MXD nanocrystal dispersion containing methylcellulose (MC) and gum arabic (GA), termed MG-MXD@NP, with a particle size of 110 nm. In vivo studies in C57BL/6 mice showed that MG-MXD@NP improved MXD delivery to the skin tissue, hair bulges, and hair bulbs, resulting in earlier and superior hair growth compared with a commercially available MXD lotion (Riup 5%, CA-MXD). These findings were consistent with the increased MXD contents observed in the hair bulge and hair bulb regions in the MG-MXD@NP-treated mice, and suggested a correlation between the efficiency of MXD delivery and efficacy of promotion of hair growth by products containing MXD. Furthermore, delivery of MXD using MG-MXD@NP was associated with elevated expression levels of CD34 and CD200, markers of hair follicle epithelial stem cells, which are crucial for promoting hair growth. Thus, it is possible that the upregulation of CD34 and CD200 in the MG-MXD@NP-treated mice reflects activation of the papilla cells and hair follicle stem cells known to be closely associated with hair growth enhancement. In conclusion, MG-MXD@NP, containing MXD nanocrystals in combination with MC and GA, exhibited a superior hair growth effect as compared with conventional MXD formulations. These findings suggest that this novel delivery method for MXD could represent a promising treatment approach for hair loss.

With the increase in life expectancy in Japan, the proportion of older adults requiring medical assistance continues to increase. Hence, the Japanese government proposed the establishment of a community-based integrated care system, aimed at ensuring housing, medical care, long-term care, prevention, and lifestyle support in a comprehensive manner by 2025. To achieve this paradigm shift, pharmacies must collaborate with their respective medical providers. Therefore, we conducted a questionnaire survey to investigate the characteristics of pharmacies with successful regional medical collaborations. An original questionnaire was sent to 1200 randomly sampled pharmacies in Tokyo, of which 350 (29.2%) responded. These were evaluated based on the implementation of regional medical collaboration. The adequacy of a regional medical collaboration system was positively related to the operational structure (e.g., staffing and information sharing within the pharmacy) and the number of hospitals (including clinics) issuing prescriptions. In addition, insufficient personnel and a lack of interaction with other medical providers were identified as major obstacles to implementing regional collaboration. Notably, the geographic distance from medical providers and the type of division of labor with medical providers were not associated with the adequacy of engagement in a regional medical collaboration system. The results of this study suggest that it may be possible to move closer to the realization of a community-based integrated care system through communication with other healthcare professionals and appropriate staffing regardless of the pharmacy’s business form.

Excessive inflammatory responses to viral infections, known as cytokine storms, are caused by overactivation of endolysosomal Toll-like receptors (TLRs) (TLR3, TLR7, TLR8, and TLR9) and can be lethal, but no specific treatment is available. Some quinoline derivatives with antiviral activity were tried during the recent coronavirus disease 2019 (COVID-19) pandemic, but showed serious toxicity, and their efficacy for treating viral cytokine storms was not established. Here, in order to discover a low-toxicity quinoline derivative as a candidate for controlling virally induced inflammation, we synthesized a series of derivatives of amodiaquine (ADQ), a quinoline approved as an antimalarial, and tested their effects on TLRs-mediated production of inflammatory cytokines and cell viability in vitro. In J774.1 murine macrophages, ADQ inhibited interleukin-6 (IL-6) production induced by TLR3 agonist poly(I:C), TLR7 agonist imiquimod, and TLR9 agonist cytosine-phosphate-guanosine oligodeoxynucleotide (CpG ODN) with IC₅₀ values of 2.43, 3.48, and 0.0359 µM, respectively, indicating that ADQ has a high inhibitory selectivity for TLR9 signaling. A structure–activity relationship study revealed that an appropriately substituted amino group on the phenol moiety and a halogen substituent on quinoline are important for potent anti-inflammatory activity and low cytotoxicity. ADQ analogs bearing N-butylethyl, N-3-fluoropiperidinyl, and N-4-fluoropiperidinyl groups in place of the N-diethyl group exhibited more potent activity and lower cytotoxicity than ADQ. ADQ and its analogs appear to inhibit the activity of TLRs recognizing pathogen nucleic acids via alkalinization of endolysosomes. Our results suggest that ADQ analogs are promising candidates as therapeutic agents for cytokine storms mediated by TLRs recognizing pathogen nucleic acid with reduced side effects.

Mucociliary clearance (MCC) is a host defense mechanism of the respiratory system. Beating cilia plays a crucial role in the MCC process and ciliary beat frequency (CBF) is activated by several factors including elevations of the intracellular cAMP concentration ([cAMP]_i), intracellular Ca²⁺ concentration ([Ca²⁺]_i), and intracellular pH (pH_i). In this study, we investigated whether an artichoke-extracted component cynaropicrin could be a beneficial compound for improving MCC. We found that cynaropicrin increased [cAMP]_i using A549 cells bearing Pink Flamindo. Then, we also confirmed that cynaropicrin elevates CBF using airway epithelial ciliated cells (AECCs). We next investigated the effects of cynaropicrin on the alternation of [Ca²⁺]_i, and pH_i. Cynaropicrin increased [Ca²⁺]_i, but not pH_i. Further experiments also found that cynaropicrin increased [cAMP]_i primarily by raising [Ca²⁺]_i. To elucidate the mechanisms of cynaropicrin to increase [Ca²⁺]_i, we investigated the alternation of the effects of cynaropicrin on [Ca²⁺]_i using several compounds. BTP-2 and ruthenium red (RuR) inhibited cynaropicrin-induced [Ca²⁺]_i increase and RuR reduced also [cAMP]_i. These results suggest that cynaropicrion increased [Ca²⁺]_i by augmenting the Ca²⁺ influx and that the increase of [cAMP]_i by cynaropicrin was induced by [Ca²⁺]_i elevation. Interestingly, cynaropicrin decreased the Ca²⁺ concentration in the endoplasmic reticulum following inhibition of sarco-endoplasmic reticulum Ca²⁺-ATPase (SERCA). SERCA activator CDN1163 abolished this effect. Furthermore, RuR and Ca²⁺-free conditions suppressed the increase of CBF. In conclusion, cynaropicrin inhibits SERCA, induces store-operated calcium entry, and thereby increases CBF.

This article has been deleted at the request of the authors from this journal.
Editorial Committee of the Pharmaceutical Society of Japan (February 17, 2025)

The effect of a citrus-derived flavonoid, hesperetin, on the automaticity and contraction of isolated guinea pig myocardium was examined. Hesperetin inhibited the rate of ectopic action potential firing of the pulmonary vein myocardium; the slope of the diastolic depolarization was decreased with minimum change in the action potential waveform. The effect was dependent on the concentration; the EC₅₀ value for firing rate was 56.2 ± 14.2 µM and that for the diastolic depolarization slope was 41.6 ± 13.4 µM. In the right atria, hesperetin had no effect on the beating rate at concentrations up to 30 µM, but showed a negative chronotropic effect at 100 µM. In the ventricular papillary muscles, hesperetin had no effect on the contractile force at concentrations up to 30 µM, but showed a slight positive inotropic effect at 100 µM. In isolated pulmonary vein cardiomyocytes, 30 µM hesperetin decreased the firing rate of spontaneous Ca²⁺ transients. These results showed that hesperetin has a selective inhibitory action on the pulmonary vein automaticity with no inhibitory effect on the ventricular contractile force. Clarification of the mechanism of action of hesperetin as well as further exploration of flavonoids would provide clues for the development of pulmonary vein-selective therapeutic agents.

We aimed to investigate the mechanism of high mobility group box 1 (HMGB1) in the accelerated fracture healing process during Traumatic brain injury (TBI). The lateral ventricles of mice in the TBI model group were injected with adenovirus-packaged short hairpin RNA (shRNA)-HMGB1 or overexpressing (ov)-HMGB1 vector. We found HMGB1 levels were higher in bone tissue at the fracture end of TBI combined with fracture model mice. Compared with the TBI combined with fracture model mice, the mice in the ov-HMGB1 group healed faster and the expression levels of mitochondrial autophagy-related proteins were higher. Compared to the ov-HMGB1 group, mice in the ov-HMGB1 + autophagy inhibitor cyclosporin A (CsA) and ov-HMGB1 + shRNA-phosphatase and tensin homolog-induced kinase 1 (PINK1) groups showed slower healing and lower expression of mitochondrial autophagy-associated proteins. The expression of osteocalcin (OCN), SOX9, and bone morphogenetic protein (BMP)-2 in bone tissue at the fracture end of the ov-HMGB1 + shRNA-PINK1 group was lower than that in the ov-HMGB1 group. The mRNA expression levels of chondrogenic differentiation markers in bone tissue at the fracture end of the ov-HMGB1 + shRNA-PINK1 group were lower than those in the ov-HMGB1 group. Fracture healing was accelerated during TBI, especially when HMGB1 was highly expressed, and HMGB1 promote accelerated fracture healing during TBI through PINK1/Parkin-mediated mitochondrial autophagy.

Puerarin (PU), a bioactive constituent reported to possess therapeutic effectiveness, but it suffers a drawback of poor bioavailability. In the present study, the PU nanoparticles (PU-NPs) were prepared using solvent-diffusion-evaporation method and optimized using Box–Behnken design (BBD), a response surface methodology for obtaining the optimal material ratio of PU-NPs. Further, PU and PU-NPs were evaluated to assess their cytotoxic effect and in vitro efficiency of inflammatory responses using lipopolysaccharide-sensitive macrophage cell line (RAW264.7). Also, PU-NPs were assessed for, in vivo anti-inflammatory activity using a carrageenan-induced rat paw edema model and an oral pharmacokinetic release study. PU-NPs formulation exhibited smaller particle sizes, an increase in the amorphous structure stability, and a higher dissolution rate, as compared to PU. The relative bioavailability of PU-NPs increased up to five-fold compared to PU suspension, as demonstrated by the parameters like the area under the curve (AUC), t_1/2, and the mean residence time (MRT). It mitigates enhanced cell viability and lowers the production of pro-inflammatory mediators [nitric oxide (NO), tumour necrosis factor-α (TNF-α), and interleukin-6 (IL-6)]. Moreover, PU-NPs showed a marked reduction in the development of paw edema at low doses compared to PU in an in vivo carrageenan-induced rat paw edema model. The results of the study affirm the potential of PU-NPs compared to PU in enhancing in vitro and in vivo anti-inflammatory responses by prolonging release and enhancing relative bioavailability.

Idiopathic pulmonary fibrosis (PF) is an irreversible and chronic inflammatory condition with limited therapeutic options and a high mortality rate. We aimed to determine the possible role and mechanisms of wogonin (WGN) on PF. A rat model of PF was established with intratracheally administrated with bleomycin (BLM), followed by intravenously injecting with WGN and weekly body weight measurements for four weeks. Hematoxylin–eosin (H&E) and Masson’s trichrome staining were implemented for histopathological analysis. In addition, the levels of fibrotic proteins and indicators of the mitogen-activated protein kinase (MAPK) pathway were assessed with Western blot. RT-quantitative (q)PCR experiment was conducted to investigate the fibrotic proteins’ mRNA expression. Ultimately, the concentrations of glutathione peroxidase (GSH-PX), malonaldehyde (MDA), and superoxide dismutase (SOD) were ascertained with appropriate kits. The results showed that WGN administration significantly reversed BLM-induced body weight reduction, alleviated pathological fibrosis, and reduced the Ashcroft score and the lung wet-to-dry weight ratio. Additionally, WGN suppressed the rise of fibrotic protein levels in BLM-treated rat’s lung tissues. Furthermore, WGN attenuated BLM-stimulated oxidative stress, as evidenced by the increased GSH-PX and SOD levels and decreased MDA levels in vivo. Finally, wogonin supplements significantly lowered the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK phosphorylation levels in the BLM-treated rat’s lung tissues. In conclusion, our study proved that PF induced by BLM administration can be mitigated by WGN treatment via suppressing the MAPK pathway, indicating that WGN is a candidate therapeutic agent for managing PF.

Ceramide (Cer) is synthesized in the endoplasmic reticulum (ER) using sphinganine as the common backbone and is then transported to the Golgi apparatus to synthesize two complex sphingolipids, sphingomyelin (SM) and glucosylceramide (GlcCer). Brefeldin A (BFA) affects the structure of the Golgi apparatus, resulting in the redistribution of the Golgi proteins into the ER. Therefore, BFA has been used to examine the ER-to-Golgi trafficking of lipids, but the detailed lipid changes in cells upon BFA treatment are not fully understood. Here, we examined the metabolism of deuterium-labeled sphinganine in HEK293T cells treated with BFA using liquid chromatography–tandem mass spectrometry. The BFA-pretreated cells were incubated in the presence of deuterium-labeled sphinganine and BFA for 5 min to 8 h, and the levels of dihydroceramide, Cer, dihydrosphingomyelin, SM, GlcCer, and phosphatidylcholine (PC) were investigated. The levels of Cer species in BFA-treated cells were lower than those in untreated cells within 2 h of incubation, but following >4 h of incubation, the levels of C14–20 Cer, encompassing C14–20 acyl chains, were similar in BFA-treated and untreated cells. Furthermore, BFA treatment increased the levels of C14–20 GlcCer but had little effect on those of C22–24 GlcCer. Moreover, after incubation for >4 h, BFA treatment decreased the levels of saturated C30–32 PC but had almost little effect on those of PC containing a monounsaturated C32–34 acyl chain. Our findings showed that BFA affected the metabolism of various lipids depending on the length and saturation of the fatty acid chain.