The transcriptional repressor forkhead box N3 (FOXN3) has been reported to suppress pulmonary fibrosis by inhibiting Smad transcriptional activity. However, FOXN3 becomes unstable in response to profibrotic stimuli. This study identifies poly(ADP-ribose) polymerase-1 (PARP1) as a stabilizing partner of FOXN3, preventing its degradation by blocking p38-mediated phosphorylation. Lung-specific knockout (KO) of PARP1 promotes the development of pulmonary fibrosis by reducing the abundance of FOXN3. Conditional overexpression of FOXN3 notably mitigates pulmonary fibrosis resulting from PARP1 KO by impeding Smad signaling, underscoring the critical role of the PARP1-FOXN3 axis in pulmonary fibrosis. Mechanistically, p38 is a Smad response gene that is transcriptionally repressed by the PARP1/FOXN3 complex. The disruption of PARP1 or FOXN3 increases p38 expression, which in turn facilitates FOXN3 degradation through a feedback mechanism. This cascade activates Smad signaling, leading to a profibrotic response and myofibroblast activation. Notably, levels of PARP1 and FOXN3 are significantly reduced in patients with pulmonary fibrosis, highlighting PARP1's crucial role in suppressing the disease by regulating FOXN3-mediated Smad signaling.

AIMS:To investigate whether curcumin (CUR) delivered by EGFR-targeted extracellular vesicles (EVs) alleviates asthma and to explore the underlying mechanism. MATERIALS AND METHODS:CUR was loaded into mouse bone marrow mesenchymal stem cell‑derived EVs. A bispecific fusion protein, ZEGFR‑CP05 (EGFR‑specific affibody linked to a CD63‑binding peptide), was used to modify EVs. Therapeutic efficacy and targeting were evaluated in an ovalbumin (OVA)‑induced acute asthma mouse model. RESULTS:ZEGFR‑CP05‑EVs showed enhanced lung accumulation. In asthmatic mice, ZEGFR‑CP05‑EVs‑CUR reduced airway inflammation, goblet cell hyperplasia, collagen deposition, and EGFR expression. Network pharmacology combined with experimental validation identified monoamine oxidase A (MAOA) as a novel target. Treatment suppressed MAOA expression and activity, restored serotonin (5‑HT) levels, and decreased reactive oxygen species (ROS) generation. CONCLUSIONS:ZEGFR‑CP05‑EVs mediated curcumin delivery alleviates asthma pathology by inhibiting MAOA, highlighting a promising therapeutic strategy.

Niemann-Pick disease type C1 is a rare and fatal neurodegenerative disorder caused by mutations in the NPC1 gene. Proper formation and function of vascular system are essential for maintaining homeostasis of the central nervous system. However, the vascular contribution to NPC1 pathogenesis remains largely unexplored. Here, we demonstrate that loss of Npc1, a transmembrane protein involved in intracellular cholesterol trafficking, results in vascular abnormalities in the mouse brain. Npc1-deficient mice exhibit distorted vascular architecture, increased blood-brain barrier (BBB) permeability, and enhanced microglia-endothelial cell contacts. Investigation of molecular perturbations upon endothelial Npc1 deficiency shows impairment of lipid metabolism and compromised Rap1 signaling and tight junctions in endothelial cells. These changes were in part reverted by release of lysosomal cholesterol. Collectively, these results suggest that Npc1 is involved in the regulation of brain vascular integrity and highlight brain endothelial cells as a potential therapeutic target for NPC1 disease.

Background Genetically modified mesenchymal stem cells (MSCs) have been shown to enhance their therapeutic properties, offering more effective treatment options for various diseases, including metabolic associated fatty liver disease (MASLD). The m7G methyltransferase METTL1 plays a critical role in regulating RNA splicing, stability, and translation. This study presents our findings on METTL1 modified human umbilical cord MSCs, emphasizing their therapeutic effects and the mechanisms involved in treating MASLD.Methods METTL1 knockdown MSCs were generated via lentiviral shRNA. Key characteristics, including senescence, proliferation, cell cycle, and apoptosis, were assessed in vitro. A high-fat diet (HFD)-induced MASLD mouse model was used to evaluate the effects of MSC transplantation through serological, biochemical, and pathological analyses. Molecular mechanisms were explored using immunofluorescence (IF), Western blotting (WB), and quantitative PCR (qPCR).Results Our results indicate that METTL1-deficient MSCs exhibit reduced proliferative capacity and increased susceptibility to senescence and apoptosis. Remarkably, these MSCs significantly decreased lipid accumulation in both in vitro and in vivo MASLD models. We found that METTL1-deficient MSCs secrete higher levels of NAMPT, which activates SIRT1, leading to the inhibition of SREBP1-mediated lipogenic genes. Inhibition of NAMPT reversed the protective effects of METTL1-deficient MSCs against MASLD-related lipid metabolic disorders. Furthermore, overexpression of METTL1 in MSCs exacerbated lipid metabolic disorders in MASLD mice by inhibiting the NAMPT/SIRT1/SREBP1 signaling pathway.Conclusion METTL1-deficient MSCs alleviate MASLD-associated lipid metabolic disorders via NAMPT secretion. This suggests that genetically modified MSCs targeting METTL1 may represent a promising therapeutic strategy for the treatment of MASLD.

Abstract The incidence of emotional disorders in patients with idiopathic pulmonary fibrosis (IPF) is substantially higher than that in the general population, severely compromising their quality of life. However, the underlying mechanisms remain poorly understood. In this study with multi-omics, we demonstrated that sphingosine-1-phosphate (S1P) derived from IPF lungs drive anxiety and depressive-like behaviors. Mechanistically, circulating S1P in the blood bound to hippocampal S1PR1 to regulate the PI3K/PKA/CREB signaling pathway, leading to synapse damage, the activation of microglia and astrocytes, neuroinflammation and ferroptosis in the hippocampus. Pharmacological inhibition of Sphk1, a key enzyme in S1P synthesis, reduced serum S1P levels and alleviated IPF-induced anxiety and depressive-like behaviors. Similarly, selective inhibition of hippocampal S1P receptor signaling using Fingolimod also attenuated neuroinflammation and ferroptosis and ameliorated mood disorders in IPF models. Collectively, these findings demonstrate that metabolite S1P from fibrotic lungs serves as a mediator of lung-to-brain functional influence, providing new insights into the IPF comorbid mood disorders and potential therapeutic targets.

BACKGROUND AND AIMS:RNF186, which encodes a ring-finger domain-containing E3 ubiquitin-protein ligase, has previously been implicated in the regulation of lipid metabolic disorders associated with metabolic dysfunction-related fatty liver disease (MAFLD). However, the precise mechanism by which RNF186 influences glucose metabolism in the context of MAFLD remains unclear. In this study, we aimed to elucidate the role of RNF186 in the regulation of glucose metabolism, with a particular focus on skeletal muscle. METHODS:In vitro, we treated skeletal myocytes and hepatocytes with high glucose concentrations to study the expression of RNF186 and its effects on glucose uptake and insulin signaling. In vivo, we developed a MAFLD model through long-term high-fat feeding and assessed the impact of RNF186 deficiency on glucose metabolism in skeletal muscle, liver and adipose tissue using Western blotting, quantitative PCR (qPCR), and immunofluorescence. RESULTS:Our findings demonstrate that RNF186 is regulated by glucose concentration in skeletal muscle cells and hepatocytes and is sensitive to insulin in a high-glucose environment. The deletion of RNF186 increases glucose metabolism and alleviates insulin signaling disruption in the MAFLD model, affecting skeletal muscle, liver, and adipose tissue. Furthermore, in skeletal muscle, RNF186 deficiency reduces the ER stress-mediated unfolded protein response (UPR) by preventing the ubiquitination of ATF6, leading to increased transcription of GLUT4. Additionally, RNF186 deficiency promotes the membrane translocation of GLUT4 via the AKT/TBC1D4 signaling pathway. In contrast, overexpression of RNF186 decreases AKT signaling and GLUT4 expression, resulting in exacerbated disruption of glucose metabolism in MAFLD. CONCLUSIONS:RNF186 regulates glucose metabolism across multiple tissues in MAFLD, notably by playing a dual role in modulating the transcription and translocation of GLUT4 in skeletal muscle. These findings suggest that targeting the expression of RNF186 could be a potential therapeutic strategy for treating MAFLD and related metabolic disorders.

Aging-associated loss of chromatin compaction is linked to derepression of retrotransposable elements (RTEs) in mouse and human tissues. Whether such RTE transcription contributes to the microglia activation that is common in aged brains is unknown. Here, we show that DAXX, a histone chaperone and RTE repressor, is downregulated during aging, preserves microglia homeostasis and inhibits cellular senescence. Loss of Daxx in young-adult microglia drives a reactive phenotype marked by chromatin decompaction at RTEs, loss of homeostatic markers, cell cycle re-entry and behavioral changes. This state leads to DNA damage and microglial depletion, followed by replacement with DAXX-deficient/Apoehigh microglia displaying features of senescence. Sustained induction of senescence relies on promyelocytic leukemia protein, a DAXX-interacting factor and interferon target. Together, these findings highlight the importance of heterochromatin maintenance in preserving adult microglial identity and plasticity, with broader implications for brain homeostasis, healthy aging and behavior.

BACKGROUND:Niemann-Pick disease type C1 (NPC1) is caused by NPC1 gene mutations, resulting in Purkinje cell degeneration and death, glial cell activation, and progressive neurodegeneration. Menstrual blood-derived endometrial stem cells (MenSCs) have been explored as a promising tool for treating neurodegenerative diseases due to their wide range of sources, non-invasive nature, and regular collection methods. OBJECTIVES:This study aims to investigate whether MenSCs can improve neuroinflammation and apoptosis in NPC1 mutant cell (Npc1KO BV2 cell line) and mice (Npc1-/- mice), and explore their underlying mechanisms. METHODS:MenSCs were transplanted into the 4-week-old Npc1-/- mice cerebellum through stereotaxic injection, and their effects on weight, behavior, and survival were assessed. The activation of glial cells and the survival of neurons were detected by immunofluorescence technology, and the expression level of related inflammatory factors and apoptotic proteins was detected by western blotting. The transcriptome changes in cerebellum after MenSCs transplantation were analyzed by transcriptome sequencing. The mechanism by which MenSCs treat NPC1 was validated at the cellular level using the activator butyzamide. RESULTS:MenSCs transplantation could slow down the rate of weight loss and improve motor coordination in Npc1-/- mice, but had no significant improvement in lifespan. MenSCs could mitigate the activation of glial cells, alleviate neuroinflammation, reduce cell apoptosis, and increase the number of mature neurons and Purkinje cells in the cerebellum. Transcriptome analyses results indicated that the JAK/STAT signaling pathway changed across different groups. Compared with age-matched Npc1+/+ mice, the protein expression levels of P-JAK2 and P-STAT3, the ratios of P-JAK2/JAK2 and P-STAT3/STAT3 were increased in the cerebellum of 5-week-old Npc1-/- mice in the PBS group; compared with age-matched Npc1-/- mice in the PBS group, the protein expression levels of P-JAK2 and P-STAT3, the ratios of P-JAK2/JAK2 and P-STAT3/STAT3 were decreased after MenSCs transplantation. Finally, treating Npc1KO BV2 cells with butyzamide further confirmed that MenSCs relieve inflammation and apoptosis caused by Npc1 gene mutations through JAK2/STAT3 signaling pathway. CONCLUSION:Our study demonstrate that MenSCs ameliorate neuroinflammation and apoptosis in NPC1 mutant cell and mice through JAK2/STAT3 signaling pathway.

BACKGROUND:Chemoresistance remains a pivotal challenge in the clinical management of breast cancer. A systematic dissection of global research trends, evolutionary hotspots, and emerging frontiers in this field is imperative for advancing therapeutic strategies. METHODS:Relevant literature on breast cancer chemoresistance from 1994 to 2024 was retrieved from the Science Citation Index-Expanded (SCI-E) database of the Web of Science Core Collection (WoSCC). Bibliometric analysis was conducted using Co-Occurrence (COOC 14.5), CiteSpace (6.1.R6), and R software (v4.2.3; packages: bibliometrix, ggplot2, and tidyverse). RESULTS:A total of 1,929 publications involving 75 countries/regions, 2,355 institutions, and 12,046 authors were analyzed. The journal Cancers published the highest number of articles (87). China contributed the largest share (803 articles), with Nanjing Medical University (53 articles) and researchers Ma Xin and Wang Yan (11 articles each) identified as the most productive institution and authors, respectively. Keyword analysis indicated that "triple-negative breast cancer (TNBC)," "doxorubicin resistance," and "breast cancer stem cells" were long-standing core topics. In the context of the big-data era, bioinformatics-driven multi-omics analyses have become mainstream approaches for investigating chemoresistance mechanisms. Targeting ferroptosis-related pathways has emerged as a novel therapeutic strategy in this field. Burst detection revealed "prognostic biomarkers" and "liquid biopsy" as current research hotspots, while reference analysis further suggested "circular RNA" and "breast cancer stemness" as critical future research directions. CONCLUSIONS:Through multidimensional bibliometric analyses, this study elucidates the developmental trajectory and potential breakthroughs in breast cancer chemoresistance research, offering a theoretical framework and translational insights for deeper exploration of resistance mechanisms and the design of precision therapeutic strategies.

Spinal cord injury (SCI) is a common neurological trauma that cannot be completely cured with surgical techniques and medications. In this study, we established a mouse SCI model and used an adeno-associated virus (AAV) to achieve the high expression of sonic hedgehog (Shh) at the injury site to further investigate the therapeutic effect and mechanism of Shh on SCI. The results of the present study show that Shh may promote motor function recovery. The present findings demonstrate the protective effect of Shh overexpression in SCI by regulating the proliferation and apoptosis of nerve cells at the site of SCI. Shh promotes the proliferation of early microglia, inhibits the proliferation of early astrocytes, and promotes the formation of neurons at the site of injury. In addition, Shh may inhibit apoptosis at the SCI site. The mechanism by which Shh regulates nerve cells at the site of SCI may involve glioma-associated oncogene 1 (Gli1). The present research indicates that Gli1 regulates the transforming growth factor-β (TGF-β) signaling pathway, inhibiting the classic TGF-β1/Smad signaling pathway and activating the TGF-β1/extracellular regulated protein kinase (ERK) signaling pathway. Collectively, these findings suggest that Shh is a regulatory molecule involved in nerve cell proliferation and apoptosis. High Shh expression can accelerate motor function recovery after SCI, indicating that it may be a promising therapeutic approach for SCI.

Proper formation of the complex neurovascular unit (NVU) along with the blood-brain barrier is critical for building and sustaining a healthy, functioning central nervous system. The RNA binding protein argonaute2 (Ago2) mediates microRNA (miRNA)-mediated gene silencing, which is critical for many facets of brain development, including NVU development. Here, we found that Ago2 in glutamatergic neurons was critical for NVU formation in the developing cortices of mice. Glutamatergic neuron-specific loss of Ago2 diminished synaptic formation, neuronal-to-endothelial cell contacts, and morphogenesis of the brain vasculature, ultimately compromising the integrity of the blood-brain barrier. Ago2 facilitated miRNA targeting of phosphatase and tensin homolog (Pten) mRNA, which encodes a phosphatase that modulates reelin-dependent phosphatidylinositol 3-kinase (PI3K)-Akt signaling within the glutamatergic subpopulation. Conditionally deleting Pten in Ago2-deficient neurons restored Akt2 phosphorylation as well as postnatal development and survival. Several mutations in AGO2 impair small RNA silencing and are associated with Lessel-Kreienkamp syndrome, a neurodevelopmental disorder. When expressed in a neuronal cell line, these human AGO2 loss-of-function variants failed to suppress PTEN, resulting in attenuated PI3K-Akt signaling, further indicating that dysregulation of Ago2 function may contribute to both impaired development and neurological disorders. Together, these results identify Ago2 as central to the engagement of neurons with blood vessels in the developing brain.

Inflammatory bowel diseases (IBDs), including Crohn's disease (CD) and ulcerative colitis (UC) are chronic recurrent nonspecific intestinal disease. Current IBD therapeutics cannot fundamentally change the natural course of IBD. Therefore, it is of great significance to find new treatment strategies for IBD. Preclinical and clinical studies have shown that mesenchymal stem cells (MSCs) are a promising therapeutic approach. However, the mechanism by which MSCs alleviate colitis and how MSCs affect intestinal mucosal barrier is still unclear. LPS-exposed human colonic epithelial cancer cell lines Caco2 and HT29. Dextran sulfate sodium (DSS)-induced IBD mouse were treated with MenSCs. We found that LPS downregulates intercellular junction proteins and induces the production of inflammatory cytokines in intestinal epithelial cells. MenSCs reduced paracellular permeability and restored barrier integrity in Caco2 cells. In Vivo, MenSCs mitigated DSS-induced colitis in mice by reducing body weight loss, colonic shortening, and disease activity index scores and by inhibiting the expressions of the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. MenSCs increased the expression of TJ proteins, improved the destruction of tight junction (TJ) structures, and reduced intestinal epithelial permeability. Furthermore, MenSCs could inhibit NF-κB p65 phosphorylation and the expression of Snail and prevent Snail nuclear localization, thereby maintaining tight and adherens junctions. Our findings demonstrate that MenSCs alleviate intestinal inflammation and enhance barrier function by suppressing the NF-κB/Snail signaling axis, offering a promising therapeutic strategy for inflammatory bowel diseases.

OBJECTIVES:Chloroquine (CQ) has been used to treat rheumatoid arthritis and systemic lupus erythematosus, but its use in multiple sclerosis (MS) is limited by side effects and insufficient efficacy. To enhance treatment outcomes, understanding CQ's therapeutic mechanisms in MS is crucial. Thus, we administered CQ to mice with experimental autoimmune encephalomyelitis (EAE) and investigated its disease-ameliorating effects and underlying cellular mechanisms. METHODS:CQ was applied intraperitoneally six days after EAE induction, immune responses, with a focus on inflammatory and regulatory T cells, as well as dendritic cells in blood, lymph nodes, spleen, and bone marrow were analyzed by flow cytometry. RESULTS:CQ treatment significantly reduced cumulative disease score and maximal disease score in CQ-treated group. Immunohistochemical analysis of the spinal cords confirmed the reduced demyelination after CQ treatment, which is accompanied by significantly decreased infiltration of T cells, B cells, and macrophages, and less activated microglia cells. Flow cytometry analysis of peripheral lymphoid organs revealed a significant decrease of inflammatory Th17 cells, which is associated with reduced pDC and their IFN-α expression, as well as Treg cells in CQ-treated mice. Indeed, depletion of pDC alone or simultaneously with CQ treatment significantly reduced EAE severity. CONCLUSION:Our results demonstrated that CQ treatment inhibits the development of EAE disease on one hand by enhancing the expansion of Treg in dLN and spleen, and on the other hand by inhibiting the accumulation of pDC and their IFN-α expression in the spleen and bone marrow. This joint effort restricts the level of inflammation in peripheral and later in CNS. Furthermore, developing a pDC-targeted CQ treatment will not only increase the treatment efficiency, but also largely decrease side effects.

Mesenchymal stem cells (MSCs) therapy is a potential treatment strategy for ulcerative colitis (UC). The expression of the Id2 (inhibitor of differentiation factor-2) gene is closely associated with the pathogenesis and prognosis of UC. However, the role of Id2 in the therapeutic efficacy of MSCs for UC remains unclear. In this study, the MSCs that either overexpressed or knocked down the Id2 gene were employed to alleviate dextrose sodium sulfate (DSS) induced UC in mice. The results indicated that MSCs overexpressing Id2 showed no significant therapeutic advantage in MSCs for UC treatment. In contrast, MSCs with Id2 knockdown demonstrated a marked reduction in therapeutic efficacy on UC, evidenced by decreased body weight, elevated disease activity index (DAI), shortened colon length, increased histopathological damage, disruption of the colonic mucosal barrier, elevated levels of pro-inflammatory cytokines, and reduced tuft cell densities in mice. Notably, Id2 knockdown in MSCs impaired their ability to regulate intestinal microbiota in UC mice, significantly promoting the growth of potentially pathogenic bacteria such as Oscillibacter and Escherichia-Shigella, while decreasing the abundance of anti-inflammatory bacteria like Dubosiella. Transcriptome sequencing analysis revealed altered gene expression involved in immune regulation and signaling pathways (Wnt and Notch), including downregulation of CD1D, CD83, SAMHD1, PRRX1, AXIN2, JAG1, and DLL1, alongside upregulation of SPHK1. Our findings underscore the pivotal role of Id2 in the therapeutic efficacy of MSC treatments for UC.

OBJECTIVE:Emerging evidence suggests that dysregulated neuroimmune pathways are implicated in schizophrenia (SCZ); however, the mechanisms connecting early-life inflammation to adult psychiatric outcomes remain inadequately understood. This study examines the role of the pro-inflammatory alarmin S100A8, a calcium- and zinc-binding protein that significantly influences the regulation of inflammatory processes and immune responses, as a potential convergent hub in pediatric infections and SCZ. Furthermore, the study characterizes the behavioral effects of S100A8 using a novel ELNI model. METHODS:We analyzed gene expression datasets from blood and brain of SCZ patients and pediatric infections (GEO accessions: GSE53987, GSE73464, GSE38484) using limma. Genetic regulation of S100A8 was examined by integrating GWAS (PGC-SCZ3) and TWAS (PsychENCODE) data. An ELNI mouse model was established via LPS injections on postnatal days 24-30. Behavioral tests, qRT-PCR, and immunofluorescence were used to assess neuroinflammation and behavioral phenotypes. RESULTS:S100A8 was upregulated in the blood and postmortem brain tissues of SCZ patients, as well as in the blood of children with bacterial infections. The rs10908557 risk allele was associated with increased S100A8 expression. LPS-induced early-life inflammation in mice led to transient growth impairment, prepulse inhibition deficits, and depressive-like behaviors. S100A8 overexpression in the hippocampus correlated with microglial activation and impaired sensorimotor gating. CONCLUSION:S100A8 serves as a shared genetic and transcriptional biomarker between pediatric infections and SCZ. Early-life inflammation induces persistent SCZ-relevant behaviors through S100A8-mediated neuroinflammation. The ELNI model offers a translational platform for studying neurodevelopmental origins of psychiatric disorders, highlighting S100A8 as a potential biomarker and therapeutic target.

The paired box 6 (Pax6) gene encodes a highly conserved transcription factor, involved in the development of eyes, brain, and endocrine glands. Homozygous loss of Pax6 resulted in neonatal death in mice, plus loss of eyes and malformation of cerebral cortex. In patients with heterozygous Pax6 mutations, a reduction in thickness of the frontoparietal cortex was detected, which was also observed in small eye mice. In this study, we found that Pax6 overexpression increased the cortical thickness, especially in the intermediate zone of the cortex, which conflicts with the report of Manuel et al. Pax6 overexpression appears to detain neurons in the intermediate zone while promoting cell proliferation. It is worth noting that the impact of Pax6 overexpression on cortical thickness and neuronal migration was temporal, explaining the differences with other reports. We postulated that the alteration of Pax6 isoform ratio by autoregulation might be responsible for this. JASPAR analysis together with the results of qPCR, Western blot, CUT&Tag, and rescue experiments revealed that Pax6 regulates neuronal migration and cell proliferation by indirectly mediating Wnt3a expression. Therefore, we propose that Pax6 participates in corticogenesis via interaction with Wnt3a in regulating neuronal migration and cell proliferation.

Neurodegenerative diseases, such as Alzheimer's disease, Parkinson's disease, and Huntington's disease, are characterized by the progressive loss of neuronal function and structure, leading to severe morbidity and mortality. Current therapeutic approaches are ineffective at stopping or reversing disease progression. Stem cell therapy has emerged as a promising candidate in research and treatment. Mesenchymal stem cells (MSCs) are considered ideal candidates for regenerative medicine because of their high proliferation rate and multi-differentiation potential. MSCs can differentiate into neurons and glial cells, modulate immune responses, and reduce inflammation, and their exosomes can promote neural repair and regulate neuronal function; thus, MSCs offer unique advantages for treating neurodegenerative diseases. However, challenges remain in optimizing cell delivery methods, ensuring the long-term survival and integration of transplanted cells, and fully understanding their therapeutic effects. This article primarily outlines the functions of MSCs in neurodegenerative diseases, with the intention that further research will fully harness their potential and translate these findings into clinical applications, offering new hope for patients suffering from neurodegenerative diseases.

To investigate the effect of the Npc1 gene on the biological activity of Telocytes (TCs) in the liver and to provide theoretical support for further research on the biological activity of TCs. Primary liver tissue cultures (TCs) from neonatal Npc1+/+ and Npc1−/− mice were extracted and cultured using an optimized type II collagenase-digestion protocol, and subsequently purified through a differential adhesion method. The growth state of TCs in both Npc1+/+ and Npc1−/− groups was regularly observed under an inverted microscope, and the morphology of TCs under normal growth conditions was documented. The TCs were identified using scanning electron microscopy and immunofluorescence staining. To investigate the impact of the Npc1 gene on the multilineage differentiation potential of TCs, liver TCs from Npc1+/+ and Npc1−/− groups were induced with adipogenic, osteogenic, and cardiomyoblastic differentiation solutions, respectively. TCs cell surface markers such as co-expression of vimentin/CD34, vimentin/PDGF-α, and vimentin/c-Kit in Npc1+/+ and Npc1−/− groups. "Combined light and scanning electron microscopy revealed that the cellular structure of TCs from Npc1+/+ and Npc1−/− groups was mainly composed of cell bodies and Telopodes (Tps). TCs exhibited small somata with fusiform, stellate, or spindle-shaped nuclei, depending on the number of Tps. The surface of TCs cell membrane was uneven, and there was no difference in morphology between the two groups. TCs had multilineage differentiation potential, and the positive rate of TCs induced in Npc1−/− group was significantly lower than that in the Npc1+/+ group. Our findings demonstrate that NPC1 deficiency markedly attenuates hepatic TCs' multipotency of liver TCs to differentiate into adipocytes, osteoblasts, and cardiocytes, suggesting that NPC1 protein might affect the pluripotency of TCs by regulating the lipid transport pathway. This finding provides novel insights into TC-mediated mechanisms in NPC pathology and lays a theoretical foundation for regenerative medicine strategies targeting TCs.

Alveolar‒capillary barrier disruption is a hallmark of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). The contribution of necroptosis to the compromised alveolar-barrier in ALI remains unclear. Mesenchymal stem cells (MSCs) may contribute to tissue repair in ALI and ARDS. Here we evaluated the efficacy and explored the molecular mechanisms of menstrual blood-derived endometrial stem cells (MenSCs) and MenSC-derived extracellular vesicles (MenSC-EVs) in ALI-induced alveolar epithelial barrier dysfunction. Human lung epithelial cells were stimulated with endotoxin and treated with MenSCs or MenSC-EVs, and their barrier properties were evaluated. Lipopolysaccharide (LPS)-injured mice were treated with MenSCs or MSC-EVs, and the degree of lung injury and the alveolar epithelial barrier of the lung tissue were assessed. We found that MenSCs reduced lung injury and restored alveolar-barrier integrity in lung tissue. In vitro, MenSCs reduced paracellular permeability and restored barrier integrity in human lung epithelial cells. MenSC-EVs replicated all these MenSC-mediated changes. Mechanistic research revealed that MenSCs inhibited MAPK signaling and necroptosis. JNK inhibition SP600125, and ERK inhibition U0126 or inhibition of necroptosis with Nec-1 or GSK872 diminished the beneficial anti-epithelial barrier dysfunction effects of MenSCs or MenSC-EVs. Our results suggest that human menstrual blood-derived endometrial stem cells mitigate lung injury and improve alveolar barrier properties by inhibiting MAPK-mediated necroptosis through extracellular vesicles, supporting the application of MenSCs or MenSC-derived extracellular vesicles to treat ALI or ARDS.

Pulmonary fibrosis (PF) is a chronic and progressive interstitial lung disease characterized by abnormal activation of myofibroblasts and pathological remodeling of the extracellular matrix, with a poor prognosis and limited treatment options. Lung transplantation is currently the only approach that can extend the life expectancy of patients; however, its applicability is severely restricted due to donor shortages and patient-specific limitations. Therefore, the search for novel therapeutic strategies is imperative. In recent years, stem cells have shown great promise in the field of regenerative medicine due to their self-renewal capacity and multidirectional differentiation potential, and a growing body of literature supports the efficacy of stem cell therapy in PF treatment. This paper systematically summarizes the research progress of various stem cell types in the treatment of PF. Furthermore, it discusses the primary methods and clinical outcomes of stem cell therapy in PF, based on both preclinical and clinical data. Finally, the current challenges and key factors to consider in stem cell therapy for PF are objectively analyzed, and future directions for improving this therapy are proposed, providing new insights and references for the clinical treatment of PF patients.