Biochemical pharmacology最新文献

As innovative therapies for renal anemia, roxadustat and daprodustat are specifically indicated for populations with chronic kidney disease (CKD) who exhibit a significantly higher susceptibility to cardiovascular complications. Platelets are important participants in cardiovascular and cerebrovascular thrombotic diseases, and the effects of roxadustat and daprodustat on platelets have not been clarified. The study explored their modulatory effects on platelet functions (using human platelets) and antithrombotic activity in vivo (via mouse pulmonary embolism and mesenteric thrombus models). Mechanistically, thromboxane A2/cyclic adenosine monophosphate (TXA₂/cAMP) levels were measured by ELISA, and protein expression by immunoblotting. Results revealed that roxadustat and daprodustat treatment could effectively inhibit platelet functions. Roxadustat and daprodustat inhibited collagen-induced platelet aggregation ex vivo and FeCl₃-induced mesenteric arteriolar thrombosis in mice and were protective in pulmonary embolism models. Additionally, roxadustat and daprodustat caused a decrease in TXA₂ production and an increase in cAMP signaling. Western blot assay results displayed that roxadustat and daprodustat downregulated collagen-induced platelet PI3K/AKT/HIF-1α pathway. Our study revealed the pharmacological effects of roxadustat and daprodustat in inhibiting platelet activation and thrombosis. The effects may provide some insights into the physiological activity of HIF and clinical medication safety.

Dysregulation of apoptosis pathways is a defining characteristic of cancer, that has established pro-apoptotic activation as a fundamental therapeutic strategy in oncology. The B-cell lymphoma-2 (Bcl-2) family proteins serves as master regulators of mitochondrial apoptosis, with anti-apoptotic members constituting critical checkpoints in cancer cell survival. Across diverse cancer types, overexpression of anti-apoptotic Bcl-2 proteins is a universal mechanism driving cancer cells to evade apoptosis and acquire resistance to chemotherapy. However, relying solely on agents that induce the downregulation of anti-apoptotic Bcl-2 proteins has proven inefficient for cancer treatment. Notably, the observed synergy between Bcl-2 inhibitors and other anti-tumor agents supports the development of dual-targeting regimens as promising therapeutic approaches. Here, we comprehensively review the recent progress in dual-targeted apoptotic modulation, focusing on strategies that concurrently inhibit anti-apoptotic Bcl-2 family proteins and synergistic pathways. Emerging evidence demonstrates that Bcl-2/B cell lymphoma extra large (Bcl-xL)/myeloid cell leukemia 1 (Mcl-1) inhibitors (e.g., venetoclax and the derivatives) combined with p53/murine double minute 2 (MDM2) disruptors, epigenetic modifiers (e.g., histone deacetylase inhibitors), autophagy modulators, or kinases inhibitors, achieve synergistic potency. These rationally designed combination therapies effectively suppress compensatory upregulation of alternative anti-apoptotic proteins, overcome Bcl-2/Bcl-xL/Mcl-1-driven resistance and restore drug efficacy in apoptosis-deficient cancer subtypes. This paradigm shift offers substantial potential to advance precision oncology by establishing durable responses through simultaneous blockade of multiple survival axes, and carries tremendous promise in the next-generation evolution of cancer therapeutics.

Non-alcoholic fatty liver disease (NAFLD) has become the fastest-growing etiology of cirrhosis and hepatocellular carcinoma. No Food and Drug Administration (FDA)-approved pharmacotherapy currently exists, underscoring the urgent need for novel regulatory circuits that can be translated into druggable targets. Here we demonstrate that autophagic flux is severely impaired and lipid accumulation markedly exacerbated in livers of NAFLD mouse models and in hepatocytes challenged with free fatty acid (FFA). Knock-down of metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) elevated miR-690 abundance, restored autophagic flux, and attenuated intracellular lipid deposition. Consistently, silencing methyltransferase-like 3 (METTL3) decreased MALAT1, thereby increasing miR-690 and producing the same protective phenotype, whereas METTL3 over-expression elicited the opposite effects. Mechanistically, METTL3 directly bound MALAT1 and installed N6-methyladenosine (m6A) modifications that enhanced MALAT1 stability and expression. Up-regulated MALAT1 subsequently sponged miR-690, leading to its functional depletion, autophagosome-lysosome fusion blockade, and aggravated lipid retention. Collectively, the METTL3-m6A/MALAT1/miR-690 axis orchestrates autophagy and lipid homeostasis, operationalizing an "m6A-long non-coding RNA (lncRNA)-microRNA (miRNA)" regulatory paradigm in NAFLD and offering an epitranscriptomic perspective on disease pathogenesis.

The antiseizure properties of ibogalogs, including ibogaminalog (DM506), ibogainalog (IBG), and nor-IBG, were assessed in rodents using the pentylenetetrazol (PTZ)-induced seizure test. The behavioral findings indicated that ibogalogs exhibited mild acute antiseizure effects in mice, with endpoint- and time window-dependent differences between the compounds. The antiseizure effect was suppressed by volinanserin and SB242084, consistent with the involvement of 5-HT_2A and 5-HT_2C receptors. The antiseizure activity after repeated administration (7 and 14 days) of subthreshold doses of nor-IBG (3 mg/kg) or DM506 (5 mg/kg) was higher than that after acute treatment, indicating augmented efficacy. Subthreshold doses of DM506 and nor-IBG restored the impact of PTZ on monoamine levels in hippocampal tissue following repeated administration, but not after a single dose. Additionally, the influence of ibogalogs was evaluated on epileptiform discharges induced by kainic acid (KA) in the CA3 region of the hippocampus. The results showed that nor-IBG and DM506 decreased epileptiform discharges in a concentration-dependent manner. Nor-IBG activity was inhibited by volinanserin, supporting a role for the 5-HT_2AR. Functional studies have shown that ibogalogs are more potent agonists at 5-HT_2A/2CRs than at 5-HT_1A/1BRs, supporting a role for 5-HT_2A/2CRs. In conclusion, repetitive treatment with ibogalogs induced antiseizure activity in mice through 5-HT_2A/2CR activation, accompanied by normalization of PTZ-induced alterations in hippocampal monoamines. In the hippocampal CA3 subfield, ibogalogs reduced KA-induced epileptiform discharges, where nor-IBG activity was mediated by 5-HT_2AR activation.

Loss of function of the Krüppel-like zinc finger transcription factor GLI-similar 3 (GLIS3) causes polycystic kidney disease (PKD) indicating that it plays a critical role in regulating normal kidney functions. The first postnatal month is accompanied with significant changes in gene expression and represents a key period in kidney development as well as the progression of PKD. In the current study, we examined the role of GLIS3 in the regulation of eicosanoid gene expression and metabolism during this period of kidney development. Transcriptome analysis showed that several eicosanoid metabolic genes are temporally regulated with the expression of some genes decreasing (lipoxygenase and cyclooxygenase arm) and others increasing (the cytochrome P450 pathways), suggesting that these changes are part of part of normal kidney maturation. Many of these temporal changes in eicosanoid gene expression are suppressed in GLIS3-deficient kidneys, consistent with our hypothesis that loss of GLIS3 function inhibits postnatal kidney maturation. Cistrome analyses revealed that several of these eicosanoid genes are directly regulated by GLIS3 and in coordination with hepatocyte nuclear factor 1 beta (HNF1B). Additionally, LC-MS-based eicosanoid metabolomics showed increased levels of PGD₂, PGE₂, TXB₂, and LTB₄ in PND28 GLIS3-deficient polycystic kidneys, consistent with elevated mRNA expression of Ptgs1/2, Alox5ap, Lta4h, and Tbxas1. The increased urinary excretion of PGE₂ and its metabolite PGEM in GLIS3-deficient mice correlates with increased renal production and metabolism of PGE₂ and higher Slco2a1 and Hpgd mRNA expression. Elevated levels of these eicosanoid metabolites might contribute to cystogenesis, altered osmoregulation, inflammation, and fibrosis in GLIS3-deficient kidneys.

Hepatic stellate cell (HSC) activation is central to liver fibrosis. Fibroblast activation protein α (FAPα) is highly expressed in activated HSCs, yet its regulatory role remains unclear. This study investigates the function and mechanism of FAPα in HSC activation and fibrosis progression. Using TGF-β1-induced LX2 cells and CCl₄-induced mouse models, along with small-molecule inhibitors and multi-omics analyses, we found that inhibiting FAPα suppressed HSC activation, proliferation, migration, and ameliorated fibrosis. Notably, FAPα formed a functional complex with Integrin αvβ1 in activated HSCs. Dual inhibition of FAPα/Integrin αvβ1 more effectively attenuated HSC activation and fibrosis than single-agent treatment. Transcriptomic and proteomic studies revealed that the complex acts through the GPC3/FGF21 axis. This study identifies the FAPα/Integrin αvβ1 complex as a key regulator of liver fibrosis and provides a novel combinatory therapeutic strategy for anti-fibrotic drug development.

Hepatic steatosis is a frequent complication of diabetes. To investigate the role of PACS2 in glucose and lipid metabolism in diabetic mice and to determine the underlying mechanisms. PACS2^-/- mice were used to establish diabetic models by a high-fat diet combined with intraperitoneal injection of STZ(HFD/STZ). Body weight, blood glucose, serum indexes, glucose tolerance test and insulin tolerance test were assessed. HepG2 cells were treated with high glucose and palmitic acid (HGPA). To investigate the specific mechanism of PACS2 silencing, HepG2 cells were transfected with the small interfering RNA (siRNA) of PACS2 or treated with an inhibitor of the JNK signaling (SP600125). Western blot was conducted to measure the JNK signaling pathway-related proteins. Our research findings indicate an upregulation of PACS2 expression in the liver tissues of diabetic mice. HFD/STZ induced dramatic exacerbation of higher blood glucose levels, insulin resistance, hepatic steatosis, MAMs formation, mitochondrial dysfunction and overactivation of the JNK signaling pathway in liver tissues, which was reversed by downregulation of PACS2. HepG2 cells treated with siRNA-PACS2 or SP600125 resulted in the inhibition of HGPA-mediated enhancement of JNK activation and the lipogenic enzymes expression (acetyl-CoA carboxylase 1 and fatty acid synthase), as well as the promotion of PPARα and fatty acid β-oxidation (FAO)-associated CPT1A expression. Collectively, our findings demonstrate that PACS2 deficiency alleviates hepatic steatosis and insulin resistance in diabetic mice by inhibition of JNK signaling pathway in hepatocytes.

Acute lung injury (ALI) represents the most frequent complication of sepsis; however, effective drug-based interventions are still unavailable. β-sitosterol (BS) has demonstrated anti-inflammatory effects and protective properties on alveolar epithelial barriers. This study investigated the mechanism by which BS targets alveolar macrophages to attenuate sepsis-associated acute lung injury (SALI) via in vivo and in vitro experiments. Sepsis was induced in mice through cecal ligation and puncture (CLP), and BS was administered orally. An in vitro model of lipopolysaccharide (LPS)-induced MH-S cell infection validated the proposed mechanism. Macrophage polarization and mitochondrial function were assessed using flow cytometry, electron microscopy, and Western blot analysis. Results showed that BS suppressed reactive oxygen species (ROS) production and M1 macrophage polarization in LPS-stimulated MH-S cells. Mechanistically, BS promoted lysosomal degradation of dynamin-related protein 1 (DRP1) via SUMO2/3-mediated SUMOylation, preserving mitochondrial integrity and function. Transfection of MH-S cells with DRP1 plasmid abolished the BS-mediated mitochondrial protection mechanism, reducing inhibition of oxidative stress and M1 polarization. In summary, BS inhibits M1 polarization of alveolar macrophages by promoting DRP1 SUMOylation, effectively alleviating SALI in mice. These findings support BS as a potential therapeutic agent for SALI, providing a theoretical basis for clinical application.

Myocardial fibrosis (MF) is a central feature in the terminal stages of many cardiovascular diseases. Findings from a previous study that we conducted show that when exposed to perinatal nonylphenol (NP), adult male rats developed MF. However, the underlying pathogenic mechanisms that drive its development have not been well established. This study aims to determine whether exposure to NP during the pregnancy and lactation periods causes MF in adult male offspring rats, and also to investigate the role of the TGF-β1/Smads signaling pathway in NP-induced fibrosis in cardiac fibroblasts (CFs). On postnatal day 21 (PND21), we observed a dose-dependent increase in NP levels in both the hearts and serum of the offspring rats. In comparison to the blank group, the fiber structure in the NP group was notably disorganized, and there was an occurrence of collagen deposition. Furthermore, expression levels of Collagen I/III, α-SMA, TGF-β1, MMP2, and MMP9 also increased in a dose-dependent manner in the NP group and were positively correlated with the levels of NP in the heart. In comparison to the blank control group, the levels of myocardial enzyme indicators (CK, CK-MB, LDH, and α-HBDH) as well as the levels of hydroxyproline (HYP) in both the NP and model groups showed an increase at postnatal day 90 (PND90). Furthermore, there were significant variations in the homeostasis of the outer matrix and functions related to TGF-β and Smads genes. Expression levels of Collagen I/III, TGF-β1, Smad2/3/4, α-SMA, CTGF, and MMP1 were significantly upregulated, while expression levels of Smad7 and TIMP1 were significantly downregulated. In vitro, exposure to NP significantly inhibited the activity of CFs in a dose-dependent manner and promoted abnormal expression of fibrosis-related factors and proteins related to the TGF-β1/Smads signaling pathway. TGF-β receptor type I/II inhibitors (LY2109761) reversed increased levels of NP-induced HYP and partially inhibited the expression of the TGF-β1/Smads signaling pathway and its downstream factors. This study demonstrates that exposure to NP during the pregnancy and lactation periods activates the TGF-β1/Smads signaling pathway and its downstream factors, thereby initiating the development of MF in their offspring. Moreover, the TGF-β1/Smads signaling pathway was determined to be involved in NP-induced CFs fibrosis, highlighting TGF-β1 as a critical target of NP-induced MF.