JOURNAL OF CELLULAR AND MOLECULAR MEDICINE最新文献

Iloprost, a prostacyclin (PGI₂) analogue, stimulates the IP receptor (PTGIR) to interact with the Gsα β/γ complex, leading to the activation of adenylate cyclase, which enzyme produces the second messenger cAMP. Elevation in cAMP triggers intracellular signalling events and regulates a wide variety of cellular activities. Thus, we evaluated the effects of Iloprost on platelet function and apoptosis and in vivo haemostasis and thrombosis, as well as the underlying mechanisms. Firstly, we showed that Iloprost concentration-dependently inhibited agonist-induced P-selectin exposure, integrin αIIbβ3 activation, platelet aggregation, ATP release, platelet spreading, and clot retraction. Moreover, Iloprost dose-dependently inhibited FeCl₃-induced mouse mesenteric arteriole thrombosis and markedly prolonged the tail bleeding time. Iloprost also concentration-dependently inhibited mitochondrial membrane potential (ΔΨm) depolarisation and phosphatidylserine (PS) externalisation in platelets, thereby inhibiting platelet apoptosis, and Iloprost at concentrations lower than 2 nM inhibited only platelet apoptosis but not platelet function. Importantly, Iloprost at low doses markedly elevated peripheral platelet counts in GPIbα antibody-induced immune thrombocytopenia (ITP). Mechanistic studies showed that Iloprost concentration-dependently antagonised agonist-induced decline of protein kinase A (PKA) activity and elevation of cytoplasmic Ca²⁺ in platelets, thereby attenuating platelet activation and aggregation. Elevation in PKA activity inhibited dephosphorylation of proapoptotic protein BAD and reduced caspase-3 activity, thus retarding platelet apoptosis. These data demonstrate that Iloprost dose-dependently inhibits platelet function and apoptosis by elevating PKA activity. Moderate-dose Iloprost impairs haemostasis and thrombosis via suppression of platelet function, and low-dose Iloprost elevates peripheral platelet levels by inhibiting platelet apoptosis while having no effects on platelet function.

Ferroptosis plays a crucial role in the progression of abdominal aortic aneurysm (AAA). Cuproptosis, as a new mode of death, has some similarities with ferroptosis. The primary objective of this study was to develop the role of shared cuproptosis-related differentially expressed genes (CRDEGs) and ferroptosis-related differentially expressed genes (FRDEGs) in AAA. RNA sequencing and bioinformatic analyses of human AAA tissue were used to identify dihydrolipoamide dehydrogenase (DLD), which is involved in cuproptosis and ferroptosis. qRT-PCR and IHC assays further confirmed that the DLD level was substantially higher in the AAA group than in the control group. Finally, experimental verification was conducted to identify that DLD could promote the necrosis, apoptosis and mitophagy of SMCs. In summary, our research identified DLD, linked to cuproptosis and ferroptosis, as differentially expressed in AAA across human and murine samples. DLD's role in regulating SMC necrosis, apoptosis and mitophagy positions it as a potential AAA biomarker and therapeutic target, warranting further investigation for clinical applications.

Osteoporosis is a disease of bone metabolism caused by an imbalance between osteoclast-mediated bone destruction and osteoblast-mediated bone formation. Tumour necrosis factor α (TNFα) has been reported to promote osteoclast generation and inhibit osteoblast generation. Progranulin (PGRN), which has a strong anti-inflammatory effect, interacts with tumour necrosis factor receptor (TNFR). Serum and bone tissues from patients with or without osteoporosis were collected to analyse the relationship between PGRN content and bone metabolic markers. The role of TNFα and PGRN in osteoclast differentiation was explored by using RAW 264.7 cells and BMMs. MC3T3-E1 cells and BMSCs were used to observe the role of TNFα and PGRN in osteoblast differentiation. The PGRN content in the serum and bone tissues of osteoporosis patients was lower than that in the serum and bone tissues of nonosteoporosis patients. TNFα promoted osteoclast differentiation, while PGRN inhibited this effect by interacting with TNFR1. PGRN inhibited TNFα-mediated attenuation of osteoblast differentiation by interacting with TNFR1. Moreover, PGRN alone promoted osteoblast differentiation by interacting with TNFR2. Our findings reveal that PGRN can effectively inhibit TNFα-induced osteoporosis and has a certain osteogenic effect. This discovery might provide a potential target for osteoporosis treatment.

Extracellular vesicles (EVs) are micro-nanoscale biological particles encapsulated by phospholipid bilayers, which regulate cell migration, angiogenesis and tumour cell growth by transmitting various biomolecules such as nucleic acids and proteins. EVs are composed of exosomes, microparticles and apoptotic bodies. Its benefits pass through biofilms and are not degraded by various enzymes, so it can be used as a biomarker in potential diseases and has attracted much attention from researchers. Current studies have found that EVs are involved in the development of various cardiovascular diseases (CVD), such as heart failure and myocardial ischemia–reperfusion injury. In addition, stem cell-derived EVs play an important role in the diagnosis and treatment of a variety of CVD. In this review, we present the biological features of EVs, the role of EVs in various CVD, and the challenges they encounter in the treatment of CVD.

Improving gut dysbiosis and impaired gut–brain axis has been a potent therapeutic strategy for treating myocardial infarction (MI). Geniposide (GEN), a traditional Chinese medicine extract, has demonstrated substantial cardioprotective properties post-MI. Nevertheless, the effect of GEN on gut microbial, gut–brain communication, and its potential mechanism remains unclear. In this study, we initially found that GEN significantly alleviated MI-induced cardiac dysfunction from echocardiographic data and decreased myocardial fibrosis, inflammation, apoptosis and hypertrophy post-MI. Additionally, we investigated the effects of GEN on gut pathology, and observed that GEN led to a remarkable change in gut microbiota as evidenced by altering β-diversity and short-chain fatty acids (SCFAs) levels, and alleviated intestinal damage indicated by reduced inflammation and barrier permeability post-MI. Finally, our investigation into brain pathology revealed that GEN induced a remarkable inhibition in PVN inflammation and sympathetic activity following MI. Collectively, these findings imply that the cardioprotective effects of GEN against MI were mediated possibly via an improvement in the impaired gut–brain axis. Mechanically, GEN-induced increase of microbiota-derived SCFAs might be the critical factor linking gut microbiota and reduced neuroinflammation with PVN, which leads to the suppression of sympathetic activation, therefore protecting the myocardium against MI-induced damage.

Hepatocellular carcinoma (HCC) is a major contributor to cancer-related deaths globally. Although there have been improvements in identifying treating the disease, patient outcomes are still unfavourable because of the significant variation in HCC. Mitochondrial-related genes (MRGs) are crucial in tumour metabolism, cell death and immune response, emerging as potential therapeutic targets. We analysed 2030 MRGs using TCGA, GEO and HCCDB18 databases. Differentially expressed genes were identified using edgeR and limma, and enrichment analysis was performed via the clusterProfiler package. A prognostic model was built using machine learning algorithms and evaluated using LOOCV. Immune infiltration was assessed with CIBERSORT, EPIC, MCPCounter and TIMER algorithms, and drug sensitivity was analysed using the CTRP and PRISM datasets. MRG expression levels are significantly associated with worse outcomes in HCC patients outperformed conventional clinical indicators in immune response revealed that individuals at high risk exhibited weaker immune responses, characterised by reduced immune scores, and elevated levels of CD8+ T cells and macrophages. Notably, high-risk patients also displayed heightened susceptibility to chemotherapy agents such as paclitaxel and irinotecan. Abnormal MRG expression serves as a significant biomarker for HCC prognosis. The developed model accurately predicts disease progression and can guide personalised treatment, especially for immune and chemotherapeutic therapies. Further validation with broader clinical samples is needed.

Cervical ossification of the posterior longitudinal ligament (OPLL) is an ectopic ossification disorder characterised by endochondral ossification. Its aetiology remains to be fully elucidated. This study aimed to clarify its pathogenesis through RNA sequencing of primary cells cultured from patients without cervical OPLL (control, PLL) and patients with cervical OPLL (disease, OPLL). We revealed for the first time the role of GLI1 within OPLL cells. Functional experiments indicated that GLI1, acting as a pivotal mediator between the upstream Hedgehog pathway and downstream BMP pathway, influences the pathogenesis of OPLL. The positive/negative effects on osteogenic differentiation following activation/inhibition of the Hedgehog pathway can be rescued by manipulating GLI1 expression. Overexpression of GLI1 activates BMP signalling, enhancing osteogenic capacity in PLL cells, while GLI1 knockdown suppresses BMP signal transduction, attenuating osteogenic differentiation in OPLL cells. Our findings highlight the significant role of the canonical Hedgehog signalling pathway and its interaction with the BMP pathway in the pathogenesis of OPLL.

Hepatocyte-like cells (HLCs) derived from pluripotent stem cells (PSCs) or direct reprogramming are an unlimited source of human hepatocytes for biomedical applications. HLCs are used to model human diseases, develop precise drugs and establish groundbreaking regenerative cell-based therapies. Primary human hepatocytes are the gold standard for studying human liver biology and pathology. However, their widespread use is limited by their rapid dedifferentiation in vitro, reliance on transplant-rejected donor organs, poor scalability and significant batch-to-batch variations. Therefore, high-quality ‘off-the-shelf’ HLCs are needed to overcome those limitations. Basic stepwise differentiation protocols have been developed to generate HLCs from PSCs. To evaluate the quality of the in vitro generated products, HLCs have been phenotyped using various methods. This review discusses various biological assays and methods available for the robust evaluation of HLC quality, emphasising the importance of using 24-h cultured primary human hepatocytes (PHHs) as a reference standard for comparison.

Oesophageal stricture, especially circumferential lesions, is a common complication of endoscopic submucosal dissection (ESD). However, the exact mechanisms underlying its development remain unclear. Consequently, understanding tissue microenvironment changes is crucial for identifying therapeutic targets. To address this, single-cell RNA sequencing (scRNA-seq) was performed on oesophageal stricture samples and normal controls. Alterations in cellular composition were observed, particularly in epithelial, endothelial, fibroblast and immune cells. A notable increase was observed in the number of differentiating suprabasal cell_2 (DFSC_2), which displayed pro-keratinizing traits. Detailed investigations revealed augmentation in a subset of these cells, characterised by elevated FTH1 and ECM1 expression, indicating their role in epithelial remodelling. Furthermore, fibroblast heterogeneity was demonstrated, with significant activation of myofibroblasts within stricture tissues. MDK–NCL, CXCL5/6-CXCR2, and TGFA–EGFR ligand–receptor pairs were enhanced in stricture tissues, mediating epithelial–stromal interactions. This study dissected the transcriptional landscape of postoperative oesophageal stricture tissues, providing valuable insights into stricture mechanisms and potential preventive strategies.

Chemoresistance represents a major threat to the treatment of human cancers, including cholangiocarcinoma (CHOL). Aberrant epigenetic events contribute most to the progression of CHOL and chemotherapy efficacy. PHF10, one subunit of SWI/SNF complex, expressed highly in tumours that correlated with tumorigenesis. However, the roles of PHF10 in CHOL remains unclear. Here, we utilised the bioinformatic analysis to reveal that PHF10 expressed lowly in CHOL samples relative to normal tissues. Functionally, we demonstrated that PHF10 deficiency enhanced cell proliferation, migration and self-renewal capacities of CHOL cells. PHF10 ablation further enhanced the chemoresistance of CHOL cells. The transcriptome analysis revealed that PHF10-KO could notably alter several oncogenic crosstalk, including the NF-kB signalling. As the top hit, HMGB1 mRNA expressions had the sharpest increase upon PHF10 deficiency. PHF10 coordinated with Setdb1 to mediate the H3K9me3 modifications on the HMGB1 promoter to suppress its expressions. Low PHF10 relied on HMGB1 to promote the progression of CHOL cells in vitro and in vivo. Furthermore, EZH2 mediated the H3K27me3 enrichment on the PHF10 promoter region that contributes to its low expressions. Lastly, the HMGB1 inhibitor (Glycyrrhizin) decreased proliferation rate of PHF10-deleted cells in vitro and in vivo. Targeting HMGB1 rendered PHF10^low CHOL re-sensitive to chemotherapy. Collectively, this study demonstrated that PHF10 functions as a tumour suppressor in CHOL, and is a novel target to predict and overcome chemoresistance.