Qingjun Zhu

Macrophages play a crucial role in the inflammation and wound repair processes of radiation-induced lung injury (RILI). The mechanosensitive ion channel Piezo1 is upregulated during these inflammatory and wound repair processes. However, the involvement of macrophage Piezo1 in the pathogenesis of RILI remains unclear. This study aims to elucidate the regulatory role of Piezo1 in the injury and repair process in RILI and to investigate the underlying mechanisms. We established Myeloid-specific knockout of Piezo1 (Piezo1^{∆ LysM}) mice, and the mice were subjected to total-chest irradiation (15 Gy) to simulate the clinical situation. Additionally, LPS treatment was performed on bone marrow-derived macrophages in vitro. The expression of Piezo1 in RILI was analyzed using the GEO database. In Piezo1^fl/fl and Piezo1^∆LysM mice, Piezo1 and EMT markers expression were detected by immunofluorescence, RT-qPCR, and Western blotting, the concentration of inflammatory factors by ELISA, alveolar macrophages were sorted by flow cytometry. In vivo and in vitro experiments involving Myeloid Piezo1 knockout and activation of Piezo1 with the specific agonist Yoda1 were conducted to observe the effects on lung injury. Our findings revealed that Piezo1 is upregulated in lung macrophages in mice with RILI. Myeloid Piezo1 knockout provided protective effects in mice with RILI. Myeloid-specific Piezo1 deficiency alleviates inflammatory responses, manifested by the alleviation of inflammatory damage in lung tissue, changes in the concentrations of related inflammatory factors. Simultaneously, this deficiency reduces radiation-induced pulmonary fibrosis, with improved fibrosis indicators and decreased expression of EMT markers. Moreover, myeloid Piezo1 knockout inhibited paracrine-induced EMT of BEAS-2B cells by alveolar macrophages, and reduced macrophage recruitment. Mechanistically, the regulatory effects of Piezo1 on lung macrophages were activated Ca²⁺-dependent calpain signaling, which critically upregulated MCP-1/CCR2/NF-κB and endoplasmic reticulum (ER) stress-induced ATF6/CHOP signaling axis. Our findings revealed the important function of Piezo1 in RILI, knockout or pharmacological inhibition of Piezo1may serve as a promising strategy for treating RILI.

Idiopathic pulmonary fibrosis (IPF) represents a chronic, non-reversible, and irreversible interstitial lung disease with the lack of curative interventions and a poor prognosis; identifying safe and effective therapeutic agents is of paramount importance. The Ophiopogon-Ginseng herb pair, a classical traditional Chinese medicine (TCM), exerts Qi-replenishing and Yin-nourishing effects. Its active constituents Ophiopogonin D (OP-D), Ginsenoside Rg1 (Rg1), and Ginsenoside Rg3 (Rg3) have individual anti-fibrotic potential, while their synergistic effects in IPF remain to be elucidated. This study aimed to explore how OP-D-Rg1-Rg3 attenuates IPF and to clarify its possible molecular mechanisms. Bleomycin (BLM)-induced cellular senescence and transforming growth factor-β1 (TGF-β1) induce epithelial-mesenchymal transition (EMT) in A549 cells. MTT assay and RSM determined the optimal combination ratio. Cellular senescence and EMT were assessed by SA-β-Gal staining, RT-qPCR, WB, and ELISA. An IPF mouse model was established by intratracheal BLM administration, followed by treatment with the optimized OP-D-Rg1-Rg3 combination, pirfenidone (PFD), or saline for 21 days. Pulmonary structural alterations and molecular changes were then evaluated by micro-CT, HE, Masson, and molecular analyses. The results showed that the synergistic OP-D-Rg1-Rg3 combination markedly attenuated A549 cell senescence, as evidenced by reduced SA-β-Gal activity and decreased expression of p53, p21, p16, and TGF-β1-induced EMT (upregulated E-cadherin, downregulated vimentin, fibronectin, Col-I). In vivo, the combination alleviated AEC2s senescence and pulmonary EMT, improved mouse body weight and lung morphology, reduced histopathological damage, and attenuated IPF. In conclusion, the OP-D-Rg1-Rg3 combination ameliorates IPF by inhibiting AEC2s senescence and EMT, highlighting promising clinical application prospects for IPF treatment.

Background: N6-methyladenosine (m ⁶ A) is a dynamic mRNA modification influencing transcript fate and cellular identity, especially in cancer. While oncogenic roles of m ⁶ A regulators in AML cells are known, their impact on the leukemic immune microenvironment is unclear. Methods: In this study, we constructed a single-cell atlas of macrophages in AML by integrating publicly available scRNA-seq datasets from 129 patient cohorts. Data were batch-corrected using Seurat and Harmony. Macrophage subpopulations were identified, and the expression and activity of 29 m ⁶ A regulators were analyzed. Pseudotime analysis (Monocle3), cell–cell communication (CellChat), and pathway enrichment (Metascape) analyses were performed to explore m ⁶ A-related functional programs. Survival analysis was conducted using Kaplan–Meier curves. RT-qPCR was used to verify the correlation between m ⁶ A regulatory molecules and prognosis. Results: Our findings indicated that m ⁶ A regulators are associated with macrophage fate. Writer-high macrophages showed enhanced proliferation and differentiation, maintaining monocyte-like features. Eraser-high macrophages remodeled macrophage function toward an M1-like, pro-inflammatory and antigen-presenting state. Reader-high macrophages drove macrophages toward an immunosuppressive, M2-like phenotype, while m ⁶ A-deficient cells exhibit features of functional exhaustion. Survival analysis based on bulk RNA-seq data further revealed that m ⁶ A-regulated macrophage profiles were associated with distinct prognostic stratification in AML patients. RT-qPCR analysis of macrophages isolated from clinical AML samples further validated these findings, showing that patients with favorable prognosis exhibited significantly higher expression levels of erasers compared to those with poor prognosis. Conclusion: These results highlight m ⁶ A system’s role in macrophage reprogramming and suggest that targeting m ⁶ A regulators in macrophages may serve as a potential basis for prognostic stratification and a promising therapeutic strategy in AML.

Chalcone (1,3-diaryl-2-propen-1-one) is an α, β-unsaturated ketone that serves as an active constituent or precursor of numerous natural substances, exhibiting a broad spectrum of pharmacological effects. In this study, the classical Claisen–Schmidt condensation method was used to synthesize the chalcone derivative 2′,4′-dimethoxychalcone (DTC) and evaluate its pharmacological activity. By upregulating the expression of the epithelial cell marker E-cadherin and downregulating the expression of the mesenchymal cell marker vimentin, DTC was found to inhibit transforming growth factor-β1 (TGF-β1)-induced epithelial–mesenchymal transition (EMT) process in A549 cells, maintaining the cells’ epithelial-like morphology and reducing the ability of the cells to migrate. Additionally, DTC demonstrated the ability to decrease the expression levels of nitric oxide (NO), tumor necrosis factor (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in RAW264.7 cells, suggesting a possible anti-inflammatory effect. Furthermore, DTC was found to exhibit bacteriostatic activity against Staphylococcus aureus (S. aureus), Proteus vulgaris (P. vulgaris), methicillin-resistant Staphylococcus aureus (MRSA), and Candida albicans (C. albicans), indicating that this chemical may possess broad-spectrum antibacterial activity.