Archives of Pharmacal Research最新文献

Silybin has been used as a therapeutic agent in treating liver fibrosis worldwide with unclear mechanisms. In this study, mice were fed a CDAHFD for six weeks to induce liver steatosis and mild liver fibrosis. Metabolomic analysis of mouse liver mitochondria revealed that silybin reversed metabolic abnormalities and diminished succinate accumulation within the mitochondria. Lipidomic profiling revealed marked decreases in mitochondrial membrane phospholipids (PE, PS, PC, and PI) in CDAHFD-fed mice, along with a substantial reduction in cardiolipin (CL)—a critical component for succinate dehydrogenase (SDH) complex assembly. Silybin restored mitochondrial membrane phospholipids, enhanced CRLS1 expression, facilitated the assembly of SDHA and SDHB, and rejuvenated SDH activity. CRLS1 knockdown via siRNA significantly impaired SDH function, leading to mitochondrial succinate accumulation. Moreover, silybin inhibited succinate efflux by downregulating the expression of the MCT1 transporter. Conditioned medium from palmitic acid/silybin-treated hepatocytes, containing reduced succinate levels, effectively suppressed LX-2 activation. This research indicates that silybin alleviates MASH-induced liver fibrosis by inhibiting succinate generation and its extracellular release, thereby inactivating hepatic stellate cells. These results suggest that targeting succinate production or secretion may represent a promising therapeutic strategy against liver fibrosis progression.

Collagenases (MMP-1, MMP-8, and MMP-13) play significant roles in the pathophysiology of osteoarthritis. Among these proteins, MMP-13 and MMP-8 are known for their catabolic roles in the degradation of the articular cartilage matrix. Using computational studies, we had previously observed that a metabolite of curcumin, Curcumin monoglucuronide (CMG), binds to MMPs involved in cartilage matrix destruction. The purpose of this study was to confirm the ability of CMG to protect cartilage by blocking the activity of these enzymes. The ability of CMG to bind and block the activities of MMP-13 and MMP-8 was established using several physicochemical methods. First, the protective effect of CMG on MMP-mediated cartilage destruction was demonstrated using cartilage explants in vitro. Second, the in vivo efficacy of CMG was tested by comparison with BI-4394, a specific MMP-13 inhibitor, using a rat anterior cruciate ligament transection (ACLT) model. These studies demonstrated that CMG was more effective than BI-4394 at preventing cartilage degeneration. In separate in vitro studies, CMG did not affect chondrocyte proliferation or the expression of NF-κB-mediated proinflammatory genes. Together, these findings demonstrate the therapeutic potential of CMG and emphasize the importance of inhibiting both MMP-13 and MMP-8 to achieve improved clinical outcomes.

The growing epidemic of diabetes mellitus and its associated complications presents a major global health challenge. Diabetic nephropathy (DN) is a critical microvascular complication of diabetes, accounts for approximately one-third of all related cases worldwide and frequently progresses to end-stage renal disease (ESRD) and premature mortality. Extensive experimental evidence underscores the pivotal role of chronic inflammation driven by the activation of the nuclear transcription factor NF-κB in the pathogenesis of DN. Triggered by various factors including hyperglycemia, NF-κB activation leads to the expression of numerous pro-inflammatory cytokines, chemokines and cell adhesion molecules, resulting in the pathological hallmarks of DN: podocyte injury, excessive extracellular matrix accumulation, glomerulosclerosis, epithelial-mesenchymal transition, renal tubular atrophy and increased proteinuria. Consequently, NF-κB emerges as a promising therapeutic target for DN. Naturally occurring phytoconstituents, as inhibitors of the NF-κB pathway and are gaining significant attention due to their lower toxicity, enhanced safety, greater efficacy and cost-effectiveness. This review summarizes the role of NF-κB in the pathophysiology of DN and examines recent research on medicinal plants and phytoconstituents that target the NF-κB signaling pathway in both in vitro and in vivo and in silico models. Furthermore, we elucidate their mechanisms of action and evaluate their potential as effective therapeutic agents for mitigating DN-related inflammation and complications. This provides a theoretical framework for the development of novel anti-nephropathic drugs that may overcome the limitations of current medications.

The cytotoxicity of Jaspine B was evaluated across a concentration range of 0.5–50 μM, yielding a half maximal inhibitory concentration (IC₅₀) of 2.6 μM for HepG2 human hepatocellular carcinoma cells. To explore the mechanisms of Jaspine B, we investigated the expression of sphingosine kinase 1 (SphK1), a rate-limiting enzyme involved in the production of sphingosine-1-phosphate (S1P) from sphingosine, and the modulation of sphingolipids in HepG2 cells. Jaspine B reduced SphK1 expression and S1P levels in a dose-dependent manner. Additionally, flow cytometry analysis indicated that Jaspine B induces apoptosis in HepG2. In HepG2-xenografted mice, Jaspine B treatment (20 mg/kg/every other day for four weeks) reduced tumor size without significant changes in either body or organ weight. The anti-cancer effect in these mice was linked to reduced SphK1 expression in tumor tissue and lower plasma S1P levels. The concentration of Jaspine B in tumor tissue was 4.64 ± 2.05 μM, which exceeded its IC₅₀ value (2.6 μM for cytotoxicity and 1.4 μM for SphK1 inhibition), further supporting its efficacy through SphK1inhibition. The anti-cancer effect and reduced plasma S1P levels induced by Jaspine B were comparable to PF543, a selective SphK1 inhibitor, in HepG2-xenografted mice. In conclusion, this study provides in vitro and in vivo evidence that Jaspine B is a promising anti-cancer agent for hepatocellular carcinoma, acting through SphK1 inhibition, with favorable pharmacokinetic and tumor distribution properties. This study also suggested that reduced plasma S1P levels may serve as a therapeutic biomarker for SphK1 inhibitors in hepatocellular carcinoma treatment.

Osteoarthritis (OA) is the most common joint disease worldwide. Despite significant efforts byresearchers, no disease-modifying osteoarthritis drugs (DMOADs) have been approved yet. This review compares preclinical and clinical studies of promising therapeutic approaches to gain insights into the potential reasons for their failure in clinical trials. For this purpose, prime examples of different therapeutic groups, including the antioxidant NAC, senotherapeutic UBX0101, anti-inflammatory drug Anakinra®, Wnt inhibitor Lorecevivint®, chondroanabolic growth factor Sprifermin™, and various protease inhibitors, are discussed in detail. The limitations of commonly used OA animal models are elaborated to understand this failure better. Moreover, this review addresses the challenges of patient stratification into different endotypes and phenotypes, the consideration of subgrouping in clinical trials, and the lack of suitable clinical outcome parameters. In summary, this review highlights potential reasons for the high failure rate of DMOADs in clinical trials and outlines key points for future improvement.

While studies have highlighted the negative effects of certain antidiabetic agents, similar evidence for other antidiabetic agents remains limited. In this study, we aimed to analyze bone fractures and bone mineral densities (BMDs) across patients receiving antidiabetic treatments, considering both overall and specific sites. We comprehensively searched PubMed, Embase, and ClinicalTrials.gov up to March 2024 to determine the effect of antidiabetic agent use on bone health in patients with type 2 diabetes mellitus. The primary outcome was to reveal variations in fractures across different anti-diabetic treatment modalities. The secondary outcome was the differences in BMDs based on treatment type. The fractures were grouped based on the division of specific sites. We also performed a subgroup analysis to identify differences between treatment types by dividing the study by treatment duration. The protocol is registered (CRD42024538789). A total of 234,759 individuals were enrolled in the 242 studies. We observed a trend wherein all anti-diabetic treatments were associated with decreased risk of fracture compared with placebo; however, this was not significant in direct analysis (OR 0.92, 95% CI 0.84–1.01, I² = 0, P = 0.07). For indirect analysis, glucagon-like peptide-1 receptor agonists (GLP1RAs) demonstrated a significant effect on preventing fractures in non-vertebral fracture, hip fracture, vertebral and hip fracture, and overall fracture (odds ratio [OR] 0.62, 95% confidence interval [CI] 0.43–0.90; OR 0.67, 95% CI 0.49–0.92; OR 0.64, 95% CI 0.47–0.87; OR 0.58, 95% CI 0.48–0.69, respectively). Additionally, dipeptidyl peptidase-4 inhibitors (DPP4i) significantly reduced incidences of non-vertebral fracture, vertebral and hip fracture, and overall fracture risk (OR 0.34, 95% CI 0.25–0.45; OR 0.72, 95% CI 0.55–0.95; OR 0.67, 95% CI 0.55–0.82, respectively). Meanwhile, metformin also reduces overall fracture risk (OR 0.60, 95% CI 0.42–0.88). We observed no significant differences between the antidiabetic agents according to the specific fracture site or study time point. Most antidiabetic treatments except thiazolidinediones did not increase the risk of fractures compared with the placebo. Incretin-based therapies (GLP1RA and DPP4i) exerted beneficial effects on fracture prevention compared with other treatments. These findings underscore the need for well-conducted RCTs to provide further evidence.

Parkinson’s disease (PD) is a prevalent neurodegenerative disorder marked by mitochondrial dysfunction and oxidative stress. Although levodopa remains the gold standard for managing PD motor symptoms, it lacks neuroprotective and disease-modifying effects, highlighting the need for new neuroprotective therapies. Mitophagy, the selective mitochondrial degradation by autophagy, is critical for neuronal health. Oleanolic acid, a natural hepatoprotective compound, shows uncertain efficacy in PD treatment. This study investigated the neuroprotective effects and underlying mechanisms of oleanolic acid using the 1-methyl-4-phenylpyridinium (MPP⁺)-induced cellular model and the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced mouse model of PD. In vitro, oleanolic acid demonstrated dopaminergic neuroprotection by reducing mitochondrial dysfunction and reactive oxygen species accumulation in PD cells. It upregulated the mitophagic protein DJ-1, enhancing the sequestration of damaged mitochondria into autophagosomes by mitophagy. DJ-1 knockdown attenuated oleanolic acid’s neuroprotection, confirming DJ-1’s role in oleanolic acid’s action. In vivo, pre-treatment with oleanolic acid in MPTP-induced PD mice prevented PD-like motor symptoms, reduced neuronal death in the substantia nigra, and mitigated striatal neurodegeneration. Post-treatment with oleanolic acid not only reduced these effects but also increased Bcl-2 and DJ-1 levels in the substantia nigra and striatum. In vitro, oleanolic acid activated JNK for Sp1 upregulation and nuclear translocation, which induced DJ-1 expression. Computational modeling predicted that oleanolic acid likely interacts with JNK, suggesting this binding might be necessary for JNK-Sp1-DJ-1 axis activation for mitophagy-driven neuroprotection. These results highlight oleanolic acid’s potential as a therapeutic agent in PD prevention and treatment via the JNK-Sp1-DJ-1 pathway. Further studies are required to validate its efficacy.

Saccharopolyspora spinosa, a member of the Pseudonocardiaceae family, was originally isolated from soil in the Virgin Islands and is renowned for producing spinosad, a broad-spectrum insecticidal secondary metabolite. While research on S. spinosa has historically focused on spinosad production, little is known about the broader spectrum of secondary metabolites encoded by its genome. Like Streptomyces, S. spinosa harbors numerous biosynthetic gene clusters (BGCs), many of which remain cryptic under standard laboratory conditions. In this study, the spinosyn gene cluster was deleted using the heat-sensitive vector pKC1139, generating the mutant strain S. spinosaΔSPN. The fermentation products of both the wild-type S. spinosa (B1) and S. spinosaΔSPN (B2) were analyzed through HPLC coupled with high-resolution tandem mass spectrometry (HRMS/MS). Data analysis was conducted using GNPS-based molecular networking and MestReNova. A total of seven metabolites were putatively annotated in the wild-type strain (B1), with spinosyns being the predominant compounds. In contrast, the mutant strain (B2) produced putatively linear and cyclic lipopeptides, including gageostatins and gageopeptins as the major metabolites. Additionally, the crude extract from S. spinosaΔSPN (B2) exhibited antibacterial activity, likely due to the production of lipopeptides, which are known for their antimicrobial properties. These findings indicate that deletion of the spinosyn gene cluster can activate cryptic biosynthetic pathways, leading to the discovery of novel bioactive compounds with potential applications in medicine.

The use of tacrolimus (TAC), a critical immunosuppressant post transplantation, is complicated by its high pharmacokinetic variability. While the gut microbiota has gained attention as a potential contributor, few studies have assessed its role in TAC metabolism variability. This study investigated the associations between the gut microbiota and TAC metabolism rates in kidney transplant recipients during the first month post transplantation—a crucial period for adjusting TAC to achieve therapeutic levels. We recruited 20 kidney transplant recipients and profiled their gut microbiota diversity and composition from stool samples collected before transplantation and at weeks 1 and 4 post transplantation via 16S rRNA sequencing. The TAC pharmacokinetic parameters were also collected. Associations between TAC metabolism status or pharmacokinetic parameters and gut microbiota diversity and composition were evaluated. Recipients with a fast TAC metabolism rate (C₀/D ratio < 1.05 ng/mL × 1/mg) presented significantly greater changes in both bacterial alpha and beta diversity metrics at 1 week post transplantation than did those with a slow metabolism rate (C₀/D ratio ≥ 1.05 ng/mL × 1/mg). Compared with slow metabolizers, fast metabolizers were associated with a significant increase in the abundance of three bacterial genera (Faecalibacterium, Clostridia vadinBB60, and Ruminococcus) and a significant decrease in the abundance of two bacterial species (Bacteroides plebeius and Parabacteroides goldsteinii). This study revealed links between gut microbiota diversity and composition and TAC metabolism rates in kidney transplant recipients during the early posttransplant period, underscoring the importance of investigating the gut microbiota as a contributor to TAC pharmacokinetic variability. Clarifying this causal relationship could better predict inter- and intraindividual TAC pharmacokinetic variability.

Chitinase 3-like 1 (CHI3L1) has been implicated in the pathogenesis of various diseases, including cancer. In our previous study, we found that anti-CHIL1 antibody inhibited lung tumorigenesis. It has been reported that CHI3L1 is highly overexpressed in colon cancer tissue compared with normal tissue, and high levels of serum CHI3L1 have been associated with worse colon cancer prognosis. We investigated the anticancer effect of an anti-CHI3L1 antibody on colon cancer cells. The anti-CHI3L1 antibody inhibited the cell growth of colon cancer cells in a concentration-dependent manner. The anti-CHI3L1 antibody also reduced the migration but increased apoptotic cell death in colon cancer cells. Using STRING (Search Tool for the Retrieval of Interacting Genes/Proteins), we identified an association between VEGFA and CHI3L1 in colon cancer. We confirmed interaction between VEGFA and CHI3L1 through immunoprecipitation. Furthermore, the combination treatment of the anti-CHI3L1 antibody and VEGFA siRNA inhibited cell growth but increased apoptotic cell death. Additionally, using the Human Base database, we found that CHI3L1 and VEGFA are associated with nicotinamide phosphoribosyltransferase (NAMPT). Furthermore, combining the anti-CHI3L1 antibody and NAMPT siRNA more effectively reduced cell growth and the expression of CHI3L1, VEGFA, and cell growth-related proteins, but significantly increased apoptosis-related proteins. The combination of VEGFA siRNA and NAMPT siRNA more effectively inhibited cell growth. Anti-CHI3L1 antibody inhibited the production of ATP and NADH in colon cancer and had a higher inhibitory effect on these levels when combined with NAMPT siRNA These data demonstrated that anti-CHI3L1 antibody is useful as a potential therapy for colon cancer by inhibiting NAMPT-dependent VEGFA expression and ATP and NADH levels.