关键词： Activating transcription factor 3   Osteogenic differentiation   Ferroptosis   Nrf2/HO-1   Human periodontal stem cells   Periodontitis  

摘要： Activating transcription factor 3 (ATF3) has been identified as a regulator associated with osteoblast differentiation. However, the effects of ATF3 on the osteogenic differentiation and proliferation of human periodontal stem cells (hPDLSCs) in periodontitis have not been reported. With the purpose of establishing an in vitro model of periodontitis, hPDLSCs were challenged with lipopolysaccharide (LPS). The Cell Counting Kit-8 assay was applied to assess cell viability, while reverse transcription-quantitative PCR and western blotting were employed to detect ATF3 expression. Inflammatory release was assessed using ELISA, together with western blotting. Lipid peroxidation was explored using the C11 BODIPY 581/591 probe, biochemical kits, thiobarbituric acid reactive substances (TBARS) assay and DCFH-DA staining. Iron and Fe2+ levels, and the expression levels of ferroptosis-related proteins were measured using corresponding kits and western blotting. Osteogenic differentiative capability was evaluated using alkaline phosphatase staining, Alizarin red staining and western blotting. The expression levels of proteins associated with Nrf2/HO-1 signaling were identified using western blotting. The results indicated that ATF3 expression was upregulated in LPS-induced hPDLSCs. The knockdown of ATF3 alleviated the LPS-induced inflammatory response in hPDLSCs, together with increased levels of TNF-alpha, IL-6, IL-1 beta, Cox-2 and iNOS, and decreased levels of IL-10. ATF3 silencing also led to lower TBARS production rate, and reduced levels of reactive oxygen species, iron, Fe2+, ACSL4 and TFR1, whereas it elevated the levels of SLC7A11 and GPX4. In addition, ATF3 silencing promoted hPDLSC mineralization and cell differentiation, and elevated the levels of OCN2, RUNX2 and BMP2. Additionally, ATF3 depletion upregulated the expression levels of proteins related with Nrf2/HO-1 signaling. The Nrf2 inhibitor ML385 partially counteracted the effects of ATF3 interference on the LPS

关键词： Endothelin-converting enzyme-like 1   Brachial plexus root avulsion   Motor neurons survival   c-Jun   ATF3  

摘要： Brachial plexus root avulsion (BPRA) injury arises from challenging delivery during childbirth, sports-related incidents, or car accidents, leading to extensive loss of motor neurons (MNs) and subsequent paralysis, including both motor and sensory impairment. Surgical nerve re-implantation cannot effectively restore motor function, and the survival of injured MNs is vital for axon regeneration and re-innervating the target muscles. Therefore, identifying novel molecular targets to improve injured MNs survival is of great significance in the treatment of BPRA injuries. Endothelin-converting enzyme-like 1 (ECEL1), a membrane-bound metallopeptidase, was initially identified as a molecule associated with nerve injuries. Damaged neurons exhibit a significant increase in the expression of ECEL1 following various types of nerve injuries, such as optic nerve injury and sciatic nerve injury. This study aimed to investigate the relationship between ECEL1 overexpression and the survival of injured MNs following BPRA injury. Our results observed a significant elevation in ECEL1 expression in injured MNs and positively correlated with MNs survival following BPRA injury. The transcription of ECEL1 is regulated by the transcription factors c-Jun and ATF3 in the context of BPRA injury, which is consistent with previous other nerve injuries study. In addition, the expression of TrkA gradually decreases in ECEL1-positive MNs and ECEL1 possibly preserves the activity of downstream AKT-GSK3 beta pathway of TrkA in injured MNs. In conclusion, our results introduce a promising therapeutic molecular target to assist re-implantation surgery for the treatment of BPRA injury.

关键词： Macrophages  

摘要： Metabolic dysfunction-associated steatohepatitis (MASH) is regulated by complex interplay between the macrophages and surrounding cells in the liver. Here, we show that Atf3 regulates glucose-fatty acid cycle in macrophages attenuates hepatocyte steatosis, and fibrogenesis in hepatic stellate cells (HSCs). Overexpression of Atf3 in macrophages protects against the development of MASH in Western diet-fed mice, whereas Atf3 ablation has the opposite effect. Mechanistically, Atf3 improves the reduction of fatty acid oxidation induced by glucose via forkhead box O1 (FoxO1) and Cd36. Atf3 inhibits FoxO1 activity via blocking Hdac1-mediated FoxO1 deacetylation at K242, K245, and K262 and increases Zdhhc4/5-mediated CD36 palmitoylation at C3, C7, C464, and C466;furthermore, macrophage Atf3 decreases hepatocytes lipogenesis and HSCs activation via retinol binding protein 4 (Rbp4). Anti-Rbp4 can prevent MASH progression that is induced by Atf3 deficiency in macrophages. This study identifies Atf3 as a regulator of glucose-fatty acid cycle. Targeting macrophage Atf3 or Rbp4 may be a plausible therapeutic strategy for MASH.

关键词： Pulmonary fibrosis   C-phycocyanin   lncIAPF   ATF3   Smad3   Autophagy  

摘要： Pulmonary fibrosis is a chronic and irreversible progressive lung disease caused by various factors, such as age and environmental pollution. With countries stepping into an aging society and the seriousness of environmental pollution caused by global industrialization, the incidence of pulmonary fibrosis is annually increasing. However, no effective drug is available for pulmonary fibrosis treatment. C-phycocyanin (C-PC), extracted from blue-green algae, has good water solubility and antioxidation. This study elucidated that C-PC reinforces autophagy to block pulmonary fibrogenesis by inhibiting long noncoding RNA (lncRNA) biogenesis in vivo and in vitro. Cleavage under targets and release using nuclease (CUT & RUN)-PCR, co-immunoprecipitation (Co-IP), and nuclear-cytoplasmic separation experiments clarified that C-PC blocked the nuclear translocation of activating transcription factor 3 (ATF3) to prevent the binding between ATF3 and transcription factor Smad3, thereby hindering lncIAPF transcription. Human antigen R (HuR) truncation experiment and RNA binding protein immunoprecipitation (RIP) were then performed to identify the binding domain with lncIAPF in the 244-322 aa of HuR. lncIAPF exerted its profibrogenic function through the binding protein HuR, a negative regulator of autophagy. In summary, C-PC promoted autophagy via down-regulating the lncIAPF-HuR-mediated signal pathway to alleviate pulmonary fibrosis, showing its potential as a drug for treating pulmonary fibrosis. Exploring how C-PC interacts with biological molecules will help us understand the mechanism of this drug and provide valuable target genes to design new drugs.

关键词： CGRP   ATF3   Trigeminal   Pain   Sex   Corneal   Nerve Injury   Immunocytochemistry  

摘要： Photorefractive keratectomy (PRK) is a type of eye surgery that involves removal of the corneal epithelium and its associated nerves, which causes intense acute pain in most people. We used a rat model of corneal epithelium removal (corneal abrasion) to examine underlying cellular and molecular mechanisms. In this study, we used immunohistochemistry of trigeminal ganglion (TG) to assess neuronal content of CGRP and ATF3, as well as orbital tightening (OT) to assess spontaneous pain behaviors. CGRP is an important neuropeptide in pain modulation and ATF3 is often used as a nerve injury marker. We found dynamic changes in CGRP and ATF3 in TG;both increased significantly at 24 h following corneal abrasion and females had a more pronounced increase at 24 h compared to males. Interestingly, there was no sex difference in OT behaviors. Additionally, the number of cells containing either CGRP or ATF3 in each animal correlate significantly with their OT behavior at the assessed timepoint. Since CGRP increased most in females, we tested the effectiveness of Olcegepant, a CGRP antagonist, at reducing OT behaviors following corneal abrasion in female rats. Olcegepant (1 mg/kg) was given prior to and again at 24 h after abrasion but did not change OT behaviors at any time over a 1-week period. Examination of CGRP and ATF3 together in TG showed that they rarely colocalized, indicating that the cells with upregulated CGRP are distinct from those responding to epithelial nerve injury. The studies also show that underlying molecular responses may be sex specific.

关键词： alternate day fasting   atherosclerosis   cholesterol   integrated stress response   activating transcription factor 3  

摘要： Atherosclerosis is the major contributor to cardiovascular mortality *** day fasting(ADF)has gained growing atten­tion due to its metabolic ***,the effects of ADF on atherosclerotic plaque formation remain inconsistent and con­troversial in atherosclerotic animal *** present study was designed to investigate the effects of ADF on atherosclerosis in apolipoprotein E-deficient(Apoe^(-/-))***-week-old male Apoe^(-/-)mice fed with Western diet(WD)were randomly grouped into ad libitum(AL)group and ADF group,and ADF aggravated both the early and advanced atherosclerotic lesion formation,which might be due to the disturbed cholesterol profiles caused by ADF *** significantly altered cholesterol metabolism pathways and down-regulated integrated stress response(ISR)in the *** hepatic expression of activating transcription factor 3(ATF3)was suppressed in mice treated with ADF and hepatocyte-specific overexpression of Aft3 attenuated the effects of ADF on atherosclerotic plaque formation in Apoe^(-/-)***,the expression of ATF3 could be regulated by Krüppel-like factor 6(KLF6)and both the expressions of ATF3 and KLF6 were regulated by hepatic cellular ISR *** conclusion,ADF aggravates atherosclerosis progres­sion in Apoe^(-/-)mice fed on *** inhibits the hepatic ISR signaling pathway and decreases the expression of KLF6,subsequently inhibiting ATF3 *** suppressed ATF3 expression in the liver mediates the deteriorated effects of ADF on atherosclerosis in Apoe^(-/-)*** findings suggest the potentially harmful effects when ADF intervention is applied to the population at high risk of atherosclerosis.

关键词： ATF3   hepatic stellate cell   microRNA-494-3p   Schistosoma japonicum   Schistosoma japonicum protein P40  

摘要： Schistosomiasis is a parasitic disease characterized by liver fibrosis, a process driven by the activation of hepatic stellate cells (HSCs) and subsequent collagen production. Previous studies from our laboratory have demonstrated the ability of Schistosoma japonicum protein P40 (SjP40) to inhibit HSCs activation and exert an antifibrotic effect. In this study, we aimed to elucidate the molecular mechanism underlying the inhibitory effect of recombinant SjP40 (rSjP40) on HSCs activation. Using a cell model in which rSjP40 inhibited LX-2 cell activation, we performed RNA-seq analyses and identified ATF3 as the most significantly altered gene. Further investigation revealed that rSjP40 inhibited HSCs activation partly by suppressing ATF3 activation. Knockdown of ATF3 in mouse liver significantly alleviated S. japonicum-induced liver fibrosis. Moreover, our results indicate that ATF3 is a direct target of microRNA-494-3p, a microRNA associated with anti-liver fibrosis effects. rSjP40 was found to downregulate ATF3 expression by upregulating microRNA-494-3p in LX-2 cells. This downregulation led to the inhibition of the expression of liver fibrosis proteins alpha-SMA and COL1A1, ultimately alleviating liver fibrosis caused by S. japonicum.

关键词： Zinc oxide nanoparticles   Endoplasmic reticulum stress   Heart   Apoptosis   mRNA   Acrylamide  

摘要： Due to their distinctive properties, several eco-friendly metal oxide nanoparticles were assessed for their possible cardioprotective properties. Acrylamide (ACD), a pervasive chemical in food and the environment, has been linked to cardiac toxicity. Therefore, this study examined the probable protective effect of green synthesized zinc oxide nanoparticles (GS-ZNPs) against ACD-oral exposure-induced cardiac damage in rats. For 60 days, 40 male Sprague-Dawley rats were separated into four sets that orally administered distilled water, 10-mg GS-ZNP/kg b.w., 20-mg ACD/kg b.w., or GS-ZNP + ACD. Then, cardiac damage indicators comprising CPK, CK-MB, cTn, and LDH were assessed. Besides, cardiac tissues' architecture, oxidative stress indicators, and Zn content were evaluated. The mRNA expression of the ERS-related genes, including ATF3, ATF4, ATF6, XBP-1, CHOP, JNKs, and BiP, were determined. Moreover, ERS-dependent anti-apoptotic (BCL-2) and apoptotic (Caspase-3 and BAX) genes mRNA expression were analyzed. The results showed that GS-ZNP significantly alleviated the increased ACD-induced serum cardiac damage indicators, MDA tissue content, and histopathological changes. Furthermore, the ACD-induced reduction of antioxidants and Zn heart contents were significantly reestablished by GS-ZNP. Furthermore, the ACD-induced upregulation of the ERS-encoding genes and apoptotic genes was reversed by GS-ZNP. Besides, the ACD-induced BCL-2 downregulation was counteracted by GS-ZNP. Overall, GS-ZNP could be a biologically potent compound to alleviate ACD's cardiotoxic effects, possibly by controlling the ERS and apoptosis-related genes and antioxidant activity.

关键词： apoptosis   ATF3   ferroptosis   intervertebral disc degeneration   miR-874-3p   single-cell RNA sequencing  

摘要： Intervertebral disc degeneration (IVDD) severely affects the work and the quality of life of people. We previously demonstrated that silencing activation transcription factor 3 (ATF3) blocked the IVDD pathological process by regulating nucleus pulposus cell (NPC) ferroptosis, apoptosis, inflammation, and extracellular matrix (ECM) metabolism. Nevertheless, whether miR-874-3p mediated the IVDD pathological process by targeting ATF3 remains unclear. We performed single-cell RNA sequencing (scRNA-seq) and bioinformatics analysis to identify ATF3 as a key ferroptosis gene in IVDD. Then, Western blotting, flow cytometry, ELISA, and animal experiments were performed to validate the roles and regulatory mechanisms of miR-874-3p/ATF3 signalling axis in IVDD. ATF3 was highly expressed in IVDD patients and multiple cell types of IVDD rat, as revealed by scRNA-seq and bioinformatics analysis. GO analysis unveiled the involvement of ATF3 in regulating cell apoptosis and ECM metabolism. Furthermore, we verified that miR-874-3p might protect against IVDD by inhibiting NPC ferroptosis, apoptosis, ECM degradation, and inflammatory response by targeting ATF3. In vivo experiments displayed the protective effect of miR-874-3p/ATF3 axis on IVDD. These findings propose the potential of miR-874-3p and ATF3 as biomarkers of IVDD and suggest that targeting the miR-874-3p/ATF3 axis may be a therapeutic target for IVDD.

关键词： single -cell RNA sequencing   Paravertebral muscle degeneration   Muscle stem cells   EGR1   ATF3  

摘要： Background: Paravertebral muscles (PVM) degeneration (PMD) is closely associated with low back pain. Muscle stem cells (MuSCs) can promote muscle regeneration and repair. However, our understanding of the cellular composition of PVM and the key genes-mediated MuSCs functions is limited. Methods: We analyzed cells heterogeneity and identified key genes using scRNA-seq. To validate the presence of EGR1 + MuSCs, we performed Flow cytometry assays. We utilized pseudo -time, CytoTRACE, and RNA velocity to reconstruct cells differentiation trajectory. CellPhoneDB was used to identify the interactions between MuSCs and other cells. Results: We identified 22 cell clusters and 10 cell types, including MuSCs. We found EGR1 + MuSCs representing the activated subset. We observed the overall number of MuSCs decreased, while the proportion of EGR1 + MuSCs gradually increased during PMD. We demonstrated the target ATF3 of EGR1 and EGR1 were significantly upregulated in PMD, suggesting EGR1-ATF3 signaling may regulate MuSCs functions. Cell differentiation trajectory indicated EGR1 + MuSCs exhibited higher stemness. We discovered immune cells, fibroblasts, and endothelial cells may collaborate with MuSCs through specific ligand-receptor pairs to regulate MuSCs functions. Conclusion: These findings unveils the activation of EGR1-ATF3 signaling in EGR1 + MuSCs, providing new insights into the cellular and molecular mechanisms underlying PMD.