Introduction Diabetic wound healing is a highly coordinated, multi-stage process that relies critically on the pro-regenerative microenvironment. Recently, stem cell-based therapies have emerged as a promising paradigm in regenerative medicine, largely attributable to their inherent self-renewal capacity, multilineage differentiation potential, and pro-repair secretome. Despite these advances, the therapeutic potential of neural stem cells (NSCs) in cutaneous wound repair remains largely unexplored. Methods We systematically evaluated the pro-angiogenic, antioxidant, and mitochondrial modulatory effects of NSCs on endothelial cells. A bio-inspired hydrogel was designed for NSC encapsulation with favorable preliminary biocompatibility. The in vivo pro-healing efficacy of NSCs-armed hydrogel was then assessed in diabetic mice. Results NSCs enhanced angiogenesis, antioxidant capacity, and mitochondrial function in endothelial cells. The engineered hydrogel exhibited favorable preliminary biocompatibility for the encapsulation of NSCs. This platform accelerated diabetic wound healing by regulating inflammation, promoting angiogenic, and exerting neural-supportive effects. Conclusions This NSC-loaded hydrogel platform offers a promising and clinically relevant strategy for diabetic wound repair.

Diabetic wound healing is a complex process that requires precise coordination among functional cells, with endothelial cells (ECs) playing a critical role in tissue vascularization. We begin by identifying neutrophil extracellular traps (NETs) as one of critical stressors that disrupts mitochondrial homeostasis in ECs. In addition, inadequate recruitment of ECs often leads to unsatisfactory regenerative outcomes. To address these issues, we developed a composite hydrogel formulation co-encapsulating C-X-C motif chemokine ligand 12 (CXCL12) mRNA-loaded exosomes to promote pro-regenerative endothelial cell homing, and leonurine (Leo) to regulate cellular functionalities, thereby preserving endothelial function essential for neovascularization. By pairing exosomal mRNA delivery with the MS2 coat protein (MCP)-MS2 tethering system for mRNA payload multiplication, we achieved sustained CXCL12 production and cascade-amplified recruitment of CXCR4-positive cells. Moreover, Leo released from the composite hydrogel protectively rescued mitochondrial dysfunction. Further, application of this hydrogel to full-thickness skin defects led to significantly improved wound regeneration in diabetic mice. In summary, this study establishes a therapeutic "recruit-reinforce" platform based on a dual-delivery hydrogel armed with CXCL12 mRNA-enriched exosomes and Leo, thereby precisely targeting mitochondrial homeostasis under NETs stress for efficient diabetic wound repair.

The divergent fates of neutrophils play a critical yet complex role in tissue repair. However, how to harness the heterogeneity of neutrophil phenotypes in specific clinical contexts to guide the design of therapeutic materials remains poorly explored. In this study, inspired by the accumulation of apoptotic neutrophils (aNs) and exaggerated formation of neutrophil extracellular traps (NETs) in diabetic wounds, we innovatively developed nano-platform based on Interleukin 4 (IL-4) mRNA-enriched exosomes locally delivered by bisphosphonate (BP)-magnesium (Mg²⁺) coordination hydrogels. Beyond classical anti-inflammatory and glycolysis-modulating effects, we found that diabetic wound-specific accumulation of aNs surprisingly augmented the ability of our hydrogel to upregulate pro-repair genes. Moreover, the hydrogel formulations effectively rescued defective efferocytosis caused by excessive NETs, potentially through upregulating growth arrest specific 6 (Gas6) and myeloid-epithelial-reproductive tyrosine kinase (MerTK) expression. In vivo implantation of the composite hydrogels significantly accelerated the healing rate and improved the quality of skin wounds in diabetic mice. In summary, motivated by the divergent fates of neutrophils in diabetic wounds, we successfully engineered a pro-efferocytic platform based on IL-4 mRNA-primed exosomes incorporated into a coordination hydrogel, tailored specifically to promote diabetic wound repair.

Hypertrophic scars (HSs) represent a fibroproliferative disorder characterized by excessive extracellular matrix deposition. However, the underlying regulatory mechanisms involving circular RNAs (circRNAs) have not yet been fully elucidated. This study aimed to delineate the circRNA expression landscape in HS and reveal the functional roles of a specific candidate circRNA through the competing endogenous RNA (ceRNA) network. Eighty-one differentially expressed circRNAs (DEcircRNAs) were identified in five paired HS and normal skin samples by high-throughput sequencing. Functional enrichment analysis of host genes highlighted pathways involving cell proliferation and ECM-receptor interactions. CircRNA-miRNA-mRNA network analysis revealed hsa_circ_0001946 as a significantly upregulated candidate circRNA. Subsequent RT-qPCR validated its elevation in human HS tissues and hypertrophic scar fibroblasts (HSFs). Subcellular fractionation and RNase R assays confirmed its cytoplasmic localization and circular stability. Functionally, knockdown of hsa_circ_0001946 significantly inhibited HSF proliferation, migration, and collagen gel contraction. Mechanistically, dual-luciferase reporter assays demonstrated that hsa_circ_0001946 acts as a sponge for miR-514a-3p; notably, silencing hsa_circ_0001946 attenuated the pro-fibrotic effects induced by miR-514a-3p inhibition. Furthermore, miR-514a-3p was shown to directly target and suppress COL5A2. Collectively, these findings establish that hsa_circ_0001946 promotes HS development by sequestering miR-514a-3p to derepress COL5A2 expression. This study identifies the hsa_circ_0001946/miR-514a-3p/COL5A2 axis as a novel pathogenic mechanism contributing to HS formation, suggesting hsa_circ_0001946 as a promising intervention target for mitigating pathological scarring.

Background Hypertrophic scar (HS) is a severe fibrotic disease characterized by excessive fibroblast activation and extracellular matrix deposition. While previous studies have revealed the involvement of Aortic Carboxypeptidase Like Protein (ACLP) in other fibrotic diseases, the role of ACLP in HS has not been investigated. Methods Quantitative real-time PCR (qRT-PCR), Western blotting, and immunofluorescence were applied to investigate the expression and subcellular location of ACLP. Wound healing and Transwell assays were employed to evaluate the impact of ACLP on HSF migration after ACLP siRNA transfection or recombinant human ACLP protein (rACLP) stimulation. The protein expression levels of VIM, MMP2, MMP9, α-SMA, COL I, and COL III in HSFs were also analyzed. A collagen gel contraction assay was harnessed to assess the contractile ability of HSFs after ACLP siRNA transfection or rACLP treatment. Lastly, RNA-Seq was utilized to reveal the gene expression profile of HSFs following ACLP knock-down. Results ACLP expression was increased in both HS tissues and human hypertrophic scar fibroblasts (HSFs). ACLP knock-down attenuated the horizontal and vertical migration of HSFs, collagen gel contraction activity, fibroblast to myofibroblast transition (FMT), and VIM, MMP2, MMP9, α-SMA, and COL III protein expression. Conversely, rACLP stimulation promoted HSF migration, gel contraction, FMT, and associated protein expression. Additionally, ACLP knock-down downregulated the expression of two cholesterol synthesis-related genes comprising HMGCS1 and HMGCR. Conclusions Here, for the first time, we reveal that ACLP expression is upregulated during HS and that it promotes HSF migration and myofibroblast activation. Hence, ACLP may serve as a candidate biomarker for HS pathogenesis and as an intervention target for HS prevention and treatment.

Inherited and age-related retinal degenerations are the commonest causes of blindness without effective treatments. Retinal progenitor cells (RPCs), which have the multipotency to differentiate into various retinal cell types, are regarded as a promising source of cell transplantation therapy for retinal degenerative diseases. However, the self-limited expansion of RPCs causes difficulty in cell source supply and restrict its clinical treatment. In this work, we found that inhibition of microRNA-449a (miR-449a) in RPCs can promote proliferation and inhibit apoptosis of RPCs, partially through upregulating Notch signaling. Further optimization of transduction miR-449a inhibitor into RPCs by endothelial cell-derived exosomes can promote the survival of RPCs transplanted in vivo and reduce cell apoptosis in retinal degeneration mouse models. In summary, these studies have shown that exosome-miR-449a inhibitor can effectively promote the expansion of RPCs in vitro and enhance transplanted RPCs survival in vivo, which might provide a novel intervention strategy for retinal degenerations in the future.

B cell development in bone marrow (BM) is a multi-staged process involving pro-B, pre-B, immature B, and mature B cells, among which pre-B cells undergo vigorous proliferation, differentiation, apoptosis, and gene rearrangement. While several signaling pathways participate in pre-B cell development have been clarified, detailed intrinsic mechanisms regulating pre-B cell proliferation and survival have not been fully understood. In the current study, we report that miR-582 regulates pre-B cell proliferation and survival. miR-582 is enriched in pre-B cells. Deletion of miR-582 in mice expanded the BM pre-B cell population in a cell-autonomous manner as shown by competitive BM transplantation. We show that forced miR-582 overexpression inhibited pre-B cell proliferation and survival, whereas downregulation of miR-582 by siRNA significantly promoted pre-B cell proliferation and survival in vitro. We identified that Hif1α and Rictor are authentic targets of miR-582 in pre-B cells as shown by reporter assays. Moreover, miR-582 overexpression reduced the expression of Hif1α and its downstream molecule Glut1, as well as Rictor and mTORC2 activity as shown by attenuated AKT and FoxO1 phosphorylation, while miR-582 knockdown showed opposite effects. miR-582 knockdown-induced increases in pre-B proliferation and survival was abrogated by Hif1α and Rictor inhibitors. Together, miR-582 functions as a negative regulator of pre-B cell proliferation and survival by simultaneously targeting Hif1α and mTORC2 signaling that regulates metabolism in early B cell development.

目的 研究血影蛋白βⅡ（Spectrin βⅡ）在棕榈酸诱导的小鼠原代心肌细胞DNA损伤和细胞凋亡中的作用。方法 （1）Western blot检测棕榈酸处理后小鼠原代心肌细胞Spectrin βⅡ表达量变化。（2）分别用Ad-shscramble（对照组）和Ad-sh-Spectrin βⅡ（Spectrin βⅡ干涉组）的腺病毒感染小鼠原代心肌细胞，荧光显微镜下观察病毒的感染效率。利用Western blot和qPCR检测Spectrin βⅡ的干涉效率。（3）Western blot检测棕榈酸处理后Ad-sh-scramble组和Spectrin βⅡ干涉组小鼠原代心肌细胞DNA损伤指标γ-H2AX及凋亡蛋白剪切型半胱天冬酶3（cleaved-caspase 3）的表达，采用流式细胞术检测各组细胞凋亡水平。结果 （1）给予棕榈酸处理的小鼠原代心肌细胞Spectrin βⅡ蛋白水平显著降低（P＜0.05）。（2）与Ad-sh-scramble组相比，Ad-sh-Spectrin βⅡ组Spectrin βⅡ的蛋白和mRNA水平均显著降低（P＜0.05）（3）给予棕榈酸处理后，与Ad-sh-scramble组相比，Ad-sh-Spectrin βⅡ组的γ-H2AX2和cleaved-caspase 3蛋白表达显著升高（P＜0.05），并且细胞凋亡率显著升高（P＜0.05）。结论 敲低Spectrin βⅡ加重棕榈酸诱导的小鼠原代心肌细胞DNA损伤和细胞凋亡。

Epigenetic regulations on the maintenance of neural stem cells (NSCs) are complicated and far from been fully understood. Our previous findings have shown that after blocking Notch signaling in NSCs in vivo, the stemness of NSCs decreases, accompanied by the downregulated expression of miR-582-5p. In the current study, we further investigated the function and mechanism of miR-582-5p in the maintenance of NSCs in vitro and in vivo. After transfecting a mimic of miR-582-5p, the formation of neurospheres and proliferation of NSCs and intermediate progenitor cells (NS/PCs) were enhanced, and the expression of stemness markers such as Sox2, Nestin, and Pax6 also increased. The results were reversed after transfection of an inhibitor of miR-582-5p. We further generated miR-582 knock-out (KO) mice to investigate its function in vivo, and we found that the number of NSCs in the subventricular zone (SVZ) region decreased and the number of neuroblasts increased in miR-582 deficient mice, indicating reduced stemness and enhanced neurogenesis of NSCs. Moreover, RNA-sequencing and molecular biological analysis revealed that miR-582-5p regulates the stemness and proliferation of NSCs by inhibiting secretory protein FAM19A1. In summary, our research uncovered a new epigenetic mechanism that regulates the maintenance of NSCs, therefore providing novel targets to amplify NSCs in vitro and to promote neurogenesis in vivo during brain pathology and aging.

When seeded in small numbers in medium containing 10−6 M aminopterin and fetal calf serum, V79 Chinese hamster cells required dialyzable components from the serum for growth. However, the cells grew in medium containing 10−6 M aminopterin and dialyzed serum, provided that the medium was supplemented with 10−5 M hypoxanthine and sufficient 5·10−6 M) thymidine. A growth-inhibitory property of some batches of dialyzed serum was abolished on heating the serum for 30 min at 56°. Three lines of V79 cells which lacked detectable hypoxanthine-guanine phosphoribosyl transferase (HGPRT) activity were seleccted in medium containing 8-azaguanine (8-AzG). In two of these, no spontaneous reversion to the HGPRT+ phenotype was detectable, and these cells did not cooperate metabolically with HGPRT+ cells to prevent the growth of the latter in HAT medium. One of the HGPRT− lines showed a high rate of spontaneous reversion (118/105 cells) in medium containing undialyzed serum. However, in medium containing dialyzed serum the spontaneous reversion rate fell to 4105 cells, suggesting that the revertants arising in medium containing undialyzed serum were biochemically heterogeneous.

Formation of glioma stem cells (GSCs) is considered as one of the main reasons of temozolomide (TMZ) resistance in glioma patients. Recent studies have shown that tumor microenvironment-derived signals could promote GSCs formation. But the critical molecule and underlying mechanism for GSCs formation after TMZ treatment is not entirely identified. Our study showed that TMZ treatment promoted GSCs formation by glioma cells; TMZ treatment of biopsy-derived glioblastoma multiforme cells upregulated HMGB1; HMGB1 altered gene expression profile of glioma cells with respect to mRNA, lncRNA and miRNA. Furthermore, our results showed that TMZ-induced HMGB1 increased the formation of GSCs and when HMGB1 was downregulated, TMZ-mediated GSCs formation was attenuated. Finally, we showed that the effect of HMGB1 on glioma cells was mediated by TLR2, which activated Wnt/β-catenin signaling to promote GSCs. Mechanistically, we found that HMGB1 upregulated NEAT1, which was responsible for Wnt/β-catenin activation. In conclusion, TMZ treatment upregulates HMGB1, which promotes the formation of GSCs via the TLR2/NEAT1/Wnt pathway. Blocking HMGB1-mediated GSCs formation could serve as a potential therapeutic target for preventing TMZ resistance in GBM patients.

Objective:To explore the effects of laser irradiation parameters (irradiation frequency and single duration) on tear secretion, lens and retina.Methods:Thirty-six healthy guinea pigs were randomly divided into 6 groups with random number table method according to different frequency and single exposure duration of laser to the eye, namely, high frequency short time (HFST) group, high frequency long time (HFLT) group, medium frequency short time (MFST) group, medium frequency long time (MFLT) group, low frequency short time (LFST) group and low frequency long time (LFLT) group, 6 for each group.The right eyes were irradiated with 500 lx laser as experimental eyes, and the left eyes of the guinea pigs served as the control eyes.The high, medium and low irradiation frequencies were defined as 15 times, 10 times and 5 times, respectively, and the short and long period was defined as 30 seconds and 60 seconds each time, respectively.The right eyes were irradiated based on the grouping at a 10-minute interval.The tear secretion was detected by SchirmerⅠtest; lens opacity was assessed under the slit-lamp microscope; fundus photography was performed to evaluate the general morphology of retina; retinal function was evaluated by electroretinogram (ERG) record and the thickness of retinal outer nuclear layer was measured by histopathology examination.This study protocol was approved by the Medical Ethics Committee of Air Force Military Medical University (No.20181203), and the use and care of the experimental animals complied with the ARVO statement.Results:The tear secretion was 8.00(7.37, 9.00), 8.75(8.25, 9.00), 8.50(7.75, 9.50), 9.00(8.50, 9.50), 8.00(7.37, 8.75) and 8.25(7.75, 8.75) mm/5 min in the HFST group, HFLT group, MFST group, MFLT group, LFST group and LFLT group, respectively, without significant difference among the groups(χ 2=5.502, P=0.240); after laser irradiation, there were no statistically significant differences in tear secretion between the control eyes and laser-irradiated eyes in all the groups (all at P>0.05). The lenses were clear and the fundus was normal through the experimental duration in all the groups.The amplitude of ERG a-wave was significantly reduced in the HFST group in comparison with the LFST group (P<0.05), and there was no significant difference in the b-wave amplitude among the six groups (F=1.358, P=0.268). The ERG a-, b-wave amplitudes were not significantly different between the control eyes and laser-irradiated eyes in various groups (both at P>0.05). There was no significant difference in the thickness of the outer nuclear layer of retina among the HFST group, HFLT group, MFST group, MFLT group, LFST group and LFLT group (F=0.952, P=0.463). Conclusions:The 500 lx laser irradiation is safe to ocular surface and lens, but there are some injuries to retinal function, and the injury degree is related to laser irradiation frequency.

The quiescence, activation, and subsequent neurogenesis of neural stem cells (NSCs) play essential roles in the physiological homeostasis and pathological repair of the central nervous system. Previous studies indicate that transmembrane protein Ttyh1 is required for the stemness of NSCs, whereas the exact functions in vivo and precise mechanisms are still waiting to be elucidated. By constructing Ttyh1-promoter driven reporter mice, we determined the specific expression of Ttyh1 in quiescent NSCs and niche astrocytes. Further evaluations on Ttyh1 knockout mice revealed that Ttyh1 ablation leads to activated neurogenesis and enhanced spatial learning and memory in adult mice (6–8 weeks). Correspondingly, Ttyh1 deficiency results in accelerated exhaustion of NSC pool and impaired neurogenesis in aged mice (12 months). By RNA-sequencing, bioinformatics and molecular biological analysis, we found that Ttyh1 is involved in the regulation of calcium signaling in NSCs, and transcription factor NFATc3 is a critical effector in quiescence versus cell cycle entry regulated by Ttyh1. Our research uncovered new endogenous mechanisms that regulate quiescence versus activation of NSCs, therefore provide novel targets for the intervention to activate quiescent NSCs to participate in injury repair during pathology and aging.

Background Glioma stem cells (GSCs) are glioma cells with stemness and are responsible for a variety of malignant behaviors of glioma. Evidence has shown that signals from tumor microenvironment (TME) enhance stemness of glioma cells. However, identification of the signaling molecules and underlying mechanisms has not been completely elucidated. Methods Human samples and glioma cell lines were cultured in vitro to determine the effects of adenovirus (ADV) infection by sphere formation, RT-qPCR, western blotting, FACS and immunofluorescence. For in vivo analysis, mouse intracranial tumor model was applied. Bioinformatics analysis, gene knockdown by siRNA, RT-qPCR and western blotting were applied for further mechanistic studies. Results Infection of patient-derived glioma cells with ADV increases the formation of tumor spheres. ADV infection upregulated stem cell markers and in turn promoted the capacities of self-renewal and multi-lineage differentiation of the infected tumor spheres. These ADV infected tumor spheres had stronger potential to form xenograft tumors in immune-compromised mice. GSCs formation could be promoted by ADV infection via TLR9, because TLR9 was upregulated after ADV infection, and knockdown of TLR9 reduced ADV-induced GSCs. Consistently, MYD88, as well as total STAT3 and phosphorylated (p-)STAT3, were also upregulated in ADV-induced GSCs. Knockdown of MYD88 or pharmaceutical inhibition of STAT3 attenuated stemness of ADV-induced GSCs. Moreover, we found that ADV infection upregulated lncRNA NEAT1. Knockdown of NEAT1 impaired stemness of ADV-induced GSCs. Lastly, HMGB1, a damage associated molecular pattern (DAMP) that triggers TLR signaling, also upregulated stemness markers in glioma cells. Conclusion ADV, which has been developed as vectors for gene therapy and oncolytic virus, promotes the formation of GSCs via TLR9/NEAT1/STAT3 signaling.

Malformation of blood vessels represents a hallmark of cancers, but the role and regulation of vascular mural cells (vMCs), including vascular smooth muscle cells (vSMCs) and pericytes, in tumors has not been fully understood. SM22α has been identified as a marker of vSMCs. This study aims at elucidating the function and regulation of SM22α+ mural cells (SM22-MCs) in tumor stroma. Gene-modified mice with a SM22α-CreERT2 transgene were employed to deplete SM22-MCs or activate/block Notch signaling in these cells. vSMCs from mouse dorsal aorta (vSMCs-DA) were cultured in vitro. RNA-seq was used to compare gene expression profiles. qRT-PCR and western blotting were used to determine gene expression level. Immunofluorescence was used to observe morphological alterations in tumors. SM22-MCs are essential for stabilizing tumor vasculature. Notch signaling was downregulated in tumor-derived SM22-MCs and vSMCs-DA treated with cancer cell-derived conditioned medium. Notch activation in SM22-MCs normalized tumor vasculature and repressed tumor growth. On the other hand, Notch disruption aggravated abnormal tumor vasculature and promoted growth and metastasis. Gene expression profiling of vSMCs-DA showed that Notch activation enhances their contractile phenotype and suppresses their secretory phenotype, further attenuating the invasion and proliferation of tumor cells. In contrast, Notch blockade in vSMCs-DA mitigated their contractile phenotype while strengthened the secretory phenotype. SM22-MCs facilitate vessel stability in tumors, and they gain a secretory phenotype and promote tumor malignancy in the absence of Notch signaling.

Extracellular vesicles (EVs) including exosomes can serve as mediators of cell–cell communication under physiological and pathological conditions. However, cargo molecules carried by EVs to exert their functions, as well as mechanisms for their regulated release and intake, have been poorly understood. In this study, we examined the effects of endothelial cells-derived EVs on neurons suffering from oxygen-glucose deprivation (OGD), which mimics neuronal ischemia-reperfusion injury in human diseases. In a human umbilical endothelial cell (HUVEC)–neuron coculture assay, we found that HUVECs reduced apoptosis of neurons under OGD, and this effect was compromised by GW4869, a blocker of exosome release. Purified EVs could be internalized by neurons and alleviate neuronal apoptosis under OGD. A miRNA, miR-1290, was highly enriched in HUVECs-derived EVs and was responsible for EV-mediated neuronal protection under OGD. Interestingly, we found that OGD enhanced intake of EVs by neurons cultured in vitro. We examined the expression of several potential receptors for EV intake and found that caveolin-1 (Cav-1) was upregulated in OGD-treated neurons and mice suffering from middle cerebral artery occlusion (MCAO). Knock-down of Cav-1 in neurons reduced EV intake, and canceled EV-mediated neuronal protection under OGD. HUVEC-derived EVs alleviated MCAO-induced neuronal apoptosis in vivo. These findings suggested that ischemia likely upregulates Cav-1 expression in neurons to increase EV intake, which protects neurons by attenuating apoptosis via miR-1290.

Mammalian neural stem cells (NSCs) are not only responsible for normal development of the central nervous system (CNS), but also participate in brain homeostasis and repair, thus hold promising clinical potentials in the treatment of neurodegenerative diseases and trauma. However the molecular networks regulating the stemness and differentiation of NSCs have not been fully understood. In this study, we show that Tweety-homolog 1 (Ttyh1), a five-pass transmembrane protein specifically expressed in mouse brain, is involved in maintaining stemness of murine NSCs. Blocking or activating Notch signal led to downregulation and upregulation of Ttyh1 in cultured NSCs, respectively, suggesting that Ttyh1 is under the control of Notch signaling. Knockdown of Ttyh1 in cultured NSCs resulted in a transient increase in the number and size of neurospheres, followed by a decrease of stemness as manifested by compromised neurosphere formation, downregulated stem cell markers, and increased neuronal differentiation. We generated Ttyh1 knockout mice by deleting its exon 4 using the CRISPR-Cas9 technology. Surprisingly, in contrast to a previous report, Ttyh1 knockout did not result in embryonic lethality. NSCs derived from Ttyh1 knockout mice phenocopied NSCs transfected with Ttyh1 siRNA. Immunofluorescence showed that loss of Ttyh1 leads to the increase of neurogenesis in adult mice. Taken together, these findings indicate that Ttyh1, which is likely downstream to Notch signaling, plays an important role in regulating NSCs.

长链非编码RNA (LncRNA)作为一种重要的细胞功能调控因子引起越来越多广泛的关注.它是一种长度在200-100000 nt之间的RNA分子,位于细胞核或细胞质内,不编码蛋白质.在基因组转录产物中,lncRNA所占数量比例远远超过编码RNA的比例,与DNA、RNA、蛋白质相互作用参与细胞内多种调控过程,在生命活动调控网络中有极其重要的作用.lncRNA目前是遗传学研究的热点之一.近年来许多研究表明,lncRNA在神经干细胞的自我更新、增殖和分化中均发挥十分重要的作用,同时还与许多经典信号通路交互作用,共同参与对神经干细胞的调控.目前已有许多重要意义的研究成果,带给人们更多关于lncRNA和神经干细胞的思考,也带来更多研究兴趣和方向.

Notch signaling is critically involved in neural development, but the downstream effectors remain incompletely understood. In this study, we cultured neurospheres from Nestin-Cre-mediated conditional Rbp-j knockout (Rbp-j cKO) and control embryos and compared their miRNA expression profiles using microarray. Among differentially expressed miRNAs, miR-342-5p showed upregulated expression as Notch signaling was genetically or pharmaceutically interrupted. Consistently, the promoter of the miR-342-5p host gene, the Ena-vasodilator stimulated phosphoprotein-like (Evl), was negatively regulated by Notch signaling, probably through HES5. Transfection of miR-342-5p promoted the differentiation of neural stem cells (NSCs) into intermediate neural progenitors (INPs) in vitro and reduced the stemness of NSCs in vivo. Furthermore, miR-342-5p inhibited the differentiation of neural stem/intermediate progenitor cells into astrocytes, likely mediated by targeting GFAP directly. Our results indicated that miR-342-5p could function as a downstream effector of Notch signaling to regulate the differentiation of NSCs into INPs and astrocytes commitment.