关键词： Dissolving microneedles   Melanoma   Cannabidiol   Effervescent   Tumor microenvironment   Ca2+ influx   PH   NANOPARTICLES   CHANNELS  

摘要： BackgroundConventional dissolving microneedles (DMNs) face significant challenges in anti-melanoma therapy due to the lack of active thrust to achieve efficient transdermal drug delivery and intra-tumoral *** this study, the effervescent cannabidiol solid dispersion-doped dissolving microneedles (Ef/CBD-SD@DMNs) composed of the combined effervescent components (CaCO3 & NaHCO3) and CBD-based solid dispersion (CBD-SD) were facilely fabricated by the "one-step micro-molding" method for boosted transdermal and tumoral delivery of cannabidiol (CBD).ResultsUpon pressing into the skin, Ef/CBD-SD@DMNs rapidly produce CO2 bubbles through proton elimination, significantly enhancing the skin permeation and tumoral penetration of CBD. Once reaching the tumors, Ef/CBD-SD@DMNs can activate transient receptor potential vanilloid 1 (TRPV1) to increase Ca2+ influx and inhibit the downstream NFATc1-ATF3 signal to induce cell apoptosis. Additionally, Ef/CBD-SD@DMNs raise intra-tumoral pH environment to trigger the engineering of the tumor microenvironment (TME), including the M1 polarization of tumor-associated macrophages (TAMs) and increase of T cells infiltration. The introduction of Ca2+ can not only amplify the effervescent effect but also provide sufficient Ca2+ with CBD to potentiate the anti-melanoma efficacy. Such a "one stone, two birds" strategy combines the advantages of effervescent effects on transdermal delivery and TME regulation, creating favorable therapeutic conditions for CBD to obtain stronger inhibition of melanoma growth in vitro and in *** study holds promising potential in the transdermal delivery of CBD for melanoma therapy and offers a facile tool for transdermal therapies of skin tumors.

关键词： 水牛   激活转录因子3(ATF3)   生物信息学分析   组织差异表达   功能  

摘要： 为了揭示水牛激活转录因子3(ATF3)基因的结构与功能,试验克隆了水牛ATF3基因的完整编码区(CDS)序列,采用生物信息学方法初步分析其编码产物的理化特性、结构及功能,再通过实时荧光定量PCR方法检测该基因的组织差异表达情况。结果表明:水牛ATF3基因的完整CDS序列长为546 bp,编码181个氨基酸。水牛ATF3基因的氨基酸序列与普通牛、瘤牛、牦牛、山羊和绵羊的同源性在98.3%以上。在基于核苷酸和氨基酸序列构建的系统发育树中,水牛与牛科的其他物种普通牛、瘤牛、牦牛聚集在一起。水牛ATF3蛋白属于亲水性蛋白质,没有信号肽和跨膜结构,含有亮氨基拉链(bZIP)保守结构域,定位于细胞核中。ATF3蛋白的二级结构中α-螺旋、β-转角、延伸链和无规则卷曲占比分别为51.93%、8.84%、6.08%和33.15%,三级结构与人的ATF3蛋白(5vpe.2.A)有45%的相似度。ATF3蛋白具有转录调控因子活性,通过选择性和非共价结合到顺式调控区域内特定的双链基因组DNA序列中来调节基因组的转录。水牛ATF3基因在肌肉、乳腺、心脏和肾脏中的相对表达量较高,在其余组织中低表达或几乎不表达,且该基因在泌乳期水牛乳腺中的相对表达量显著高于非泌乳期水牛(P<0.05)。说明ATF3基因可能与水牛泌乳性状有关。

关键词： 膝骨关节炎   富血小板血浆   背根神经节   激活转录因子3   疼痛  

摘要： 目的:本研究旨在观察富血小板血浆(PRP)对单碘乙酸钠(MIA)诱导的膝骨关节炎(KOA)神经病理性疼痛大鼠镇痛疗效,并对其镇痛机制进行初步探讨。方法:向大鼠左膝关节注射60 µL MIA (80 mg/mL)诱导关节退变,造模前1天及造模后第1、3、7、14天对大鼠机械痛撤足阈值、热痛撤足阈值进行测定,在造模后15、17、19天MIA + PRP组连续3天予以膝关节PRP注射治疗,MIA + NS组予以等量生理盐水注射治疗,造模后21、28天对大鼠机械痛撤足阈值、热痛撤足阈值再次进行测定,使用免疫组化染色测定背根神经节激活转录因子3 (ATF3)表达。结果:MIA造模后大鼠机械痛撤足阈值、热痛撤足阈值显著降低,予以PRP注射治疗后,MIA + PRP组机械痛撤足阈值、热痛撤足阈值显著高于MIA + NS组。造模后28天,MIA + PRP组背根神经节中ATF3蛋白表达较MIA + NS组显著降低。结论:PRP可以减轻膝骨性关节炎大鼠神经病理性痛,这一作用可能是通过减轻神经损伤实现的。

关键词： INTESTINAL MICROBIOTA   ORNITHINE LIPIDS   MEMBRANE-LIPIDS   DIET   ASSOCIATION   DIVERSITY   GENOME   ATF3   METABOLISM   EXPRESSION  

摘要： The molecular bases of how host genetic variation impacts the gut microbiome remain largely unknown. Here we used a genetically diverse mouse population and applied systems genetics strategies to identify interactions between host and microbe phenotypes including microbial functions, using faecal metagenomics, small intestinal transcripts and caecal lipids that influence microbe–host dynamics. Quantitative trait locus (QTL) mapping identified murine genomic regions associated with variations in bacterial taxa; bacterial functions including motility, sporulation and lipopolysaccharide production and levels of bacterial- and host-derived lipids. We found overlapping QTL for the abundance ofAkkermansia muciniphilaand caecal levels of ornithine lipids. Follow-up in vitro and in vivo studies revealed thatA. muciniphilais a major source of these lipids in the gut, provided evidence that ornithine lipids have immunomodulatory effects and identified intestinal transcripts co-regulated with these traits includingAtf3, which encodes for a transcription factor that plays vital roles in modulating metabolism and immunity. Collectively, these results suggest that ornithine lipids are potentially important forA. muciniphila–host interactions and support the role of host genetics as a determinant of responses to gut microbes.

关键词： Ligustrazine   Preeclampsia   Hypoxia   reoxygenation   miR-27a-3p   ATF3   Wnt   ?-catenin   PREECLAMPSIA   INVASION   PATHOPHYSIOLOGY   ATF3  

摘要： Objective: Preeclampsia (PE) is a pregnancy-specific syndrome. Ligustrazine (LSZ) is involved in hypoxia/reox-ygenation (H/R)-treated trophoblast cell regulation, but its mechanism remains elusive. This study explored the mechanism of LSZ in H/R-treated trophoblast cells to provide a theoretical basis for the new treatment method development for PE. Methods: H/R HTR8/SVneo cell model was established for PE simulation to some extent. Trophoblast cell pro-liferation, apoptosis rate, migration, and invasion were detected by MTT assay, flow cytometry, scratch test, and Transwell assay. miR-27a-3p expression in trophoblast cells was detected by RT-qPCR. Binding sites between miR-27a-3p and ATF3 were predicted using Starbase and verified by dual-luciferase reporter assay. Activating transcription factor 3 (ATF3), beta-catenin, Cyclin D1, and c-Myc protein levels were examined using Western blot. After LSZ treatment, H/R-induced HTR8/SVneo cells were delivered with miR-27a-3p mimic or ATF3 siRNA to verify their roles in HTR8/SVneo cells. Results: LSZ facilitated the proliferation, migration, and invasion of trophoblast cells and inhibited apoptosis. miR-27a-3p was elevated in H/R-induced HTR8/SVneo cells and miR-27a-3p overexpression annulled the effect of LSZ on trophoblast cells. miR-27a-3p targeted ATF3. ATF3 silencing averted the property of LSZ on tropho-blast cells. Wnt/beta-catenin pathway-related proteins were repressed in H/R-induced HTR8/SVneo cells, and LSZ activated the Wnt/beta-catenin pathway by promoting ATF3 expression. Conclusion: LSZ mediated the Wnt pathway by regulating the miR-27a-3p/ATF3 axis, thus promoting the pro-liferation and migration of trophoblast cells. The protective mechanism of LSZ showed the potential application value in the treatment of PE.

关键词： PHOSPHOENOLPYRUVATE CARBOXYKINASE   RHO-GTPASES   CANCER   EXPRESSION   CELLS   RESISTANCE   PEPCK   ATF3   STEATOHEPATITIS   TRANSCRIPTION  

摘要： Metabolic associated fatty liver disease (MAFLD) encompasses a broad spectrum of hepatic disorders, including steatosis, nonalcoholic steatohepatitis (NASH) and fibrosis. We demonstrated that phosphoenolpyruvate carboxykinase 1 (PCK1) plays a central role in MAFLD progression. Male mice with liver Pck1 deficiency fed a normal diet displayed hepatic lipid disorder and liver injury, whereas fibrosis and inflammation were aggravated in mice fed a high-fat diet with drinking water containing fructose and glucose (HFCD-HF/G). Forced expression of hepatic PCK1 by adeno-associated virus ameliorated MAFLD in male mice. PCK1 deficiency stimulated lipogenic gene expression and lipid synthesis. Moreover, loss of hepatic PCK1 activated the RhoA/PI3K/AKT pathway by increasing intracellular GTP levels, increasing secretion of platelet-derived growth factor-AA (PDGF-AA), and promoting hepatic stellate cell activation. Treatment with RhoA and AKT inhibitors or gene silencing of RhoA or AKT1 alleviated MAFLD progression in vivo. Hepatic PCK1 deficiency may be important in hepatic steatosis and fibrosis development through paracrine secretion of PDGF-AA in male mice, highlighting a potential therapeutic strategy for MAFLD. Phosphoenolpyruvate carboxykinase 1 (Pck1) is an enzyme involved in glucose production that also regulates lipogenesis and has been linked to liver steatosis. Here the authors report that deficiency of Pck1 in the liver leads to nonalcoholic fatty liver disease via activation of the RhoA/PI3K/AKT pathway in a study with male mice.

关键词： PI3K   AKT   mTOR signaling   Ewing sarcoma   ATF3   Tumor microenvironment   MOLECULAR PATHOGENESIS   CANCER-CELLS   GROWTH   NVP-BEZ235   EXPRESSION   PATHWAYS   INTERLEUKIN-8   BIOLOGY   TARGET   TUMOR  

摘要： Ewing sarcomas are aggressive pediatric tumors of bone and soft tissues driven by in frame chromosomal translocations that yield fusion proteins guiding the oncogenic program. Promising alternative strategies to ameliorate current treatments involve inhibition of the PI3K/AKT/mTOR pathway. In this study, we identified the activating transcription factor 3 (ATF3) as an important mediator of the PI3K/AKT/mTOR pathway in Ewing sarcoma cells. ATF3 exerted its pro-tumoral activity through modulation of several chemokine-encoding genes, including CXCL8. The product of CXCL8, IL-8, acts as a pro-inflammatory chemokine critical for cancer pro-gression and metastasis. We found that ATF3/IL-8 axis impacts macrophages populating the surrounding tumor microenvironment by promoting the M2 phenotype. Our study reveals valuable information on the PI3K/AKT/mTOR derived chemokine signaling in Ewing sar-coma cells: by promoting ATF3 and CXCL8 downregulation, inhibition of the PI3K/AKT/mTOR signaling pro-motes a proinflammatory response leading to upregulation of the protective anti-tumoral M1 macrophages.

关键词： Formononetin   CKD Ferroptosis   Smad3   ATF3   Nrf2   INJURY   PROTECTS   CANCER  

摘要： Aims: Chronic kidney disease (CKD) is characterized by interstitial fibrosis, while limited treatment drugs are available. Ferroptosis is a newly identified process that can trigger tubular atrophy and fibrosis. The aim of this study is to investigate the effects of formononetin (FN), a bioflavonoid, on ferroptosis and renal fibrosis. Main methods: In vivo experiments, unilateral ureteral obstruction (UUO)- and folic acid (FA, 250 mg/kg)-induced CKD models were constructed in C57BL/6 mice of 6-8 weeks old, followed by the administration with 40 mg/kg/day FN by gavage. For in vitro experiments, ferroptosis was induced with RSL3 or erastin in primary mouse renal tubular epithelial cells (TECs), followed by the addition of indicated concentrations of FN. Then, the levels of ferroptosis and fibrosis were analyzed. The translocation of Smad3, ATF3, and Nrf2 from the cytoplasm to the nucleus was checked by western blotting. The interaction of Smad3 and ATF3 was detected by Co-immunoprecipitation. Key findings: FN dramatically ameliorated tubular injury along with reduced expression of the profibrotic genes including alpha-SMA, Col1a1, and fibronectin in both two CKD mouse models and RSL3/erastin-treated TECs. Furthermore, FN administration also significantly suppressed ferroptosis, as evidenced by increased expression of SLC7A11 and GPX4, and decreased levels of 4-HNE. In mechanism, FN disrupted the interaction between Smad3 and ATF3, resulting in the blocking of their translocation from the cytoplasm to the nucleus. In addition, FN also promoted the separation of the Nrf2/Keap1 complex and enhanced Nrf2 nuclear accumulation. Significance: FN alleviates CKD by impeding ferroptosis-associated fibrosis by suppressing the Smad3/ATF3/ SLC7A11 signaling and could serve as a candidate therapeutic drug for CKD.

关键词： FATTY LIVER-DISEASE   PROTEIN-KINASE 3   DNA-DAMAGE   REGENERATION   ACTIVATION   FIBROSIS   NECROSIS   EXPRESSION   SIGNALS   ETHANOL  

摘要： Aggravation of liver steatosis shifts the hepatocellular death mode from apoptosis to necroptosis in acute and chronic liver damage. Here the authors report that the transcription factor ATF3 regulates this shift through the induction of RIPK3, a regulator of necroptosis. Hepatocellular death increases with hepatic steatosis aggravation, although its regulation remains unclear. Here we show that hepatic steatosis aggravation shifts the hepatocellular death mode from apoptosis to necroptosis, causing increased hepatocellular death. Our results reveal that the transcription factor ATF3 acts as a master regulator in this shift by inducing expression of RIPK3, a regulator of necroptosis. In severe hepatic steatosis, after partial hepatectomy, hepatic ATF3-deficient or -overexpressing mice display decreased or increased RIPK3 expression and necroptosis, respectively. In cultured hepatocytes, ATF3 changes TNF alpha-dependent cell death mode from apoptosis to necroptosis, as revealed by live-cell imaging. In non-alcoholic steatohepatitis (NASH) mice, hepatic ATF3 deficiency suppresses RIPK3 expression and hepatocellular death. In human NASH, hepatocellular damage is correlated with the frequency of hepatocytes expressing ATF3 or RIPK3, which overlap frequently. ATF3-dependent RIPK3 induction, causing a modal shift of hepatocellular death, can be a therapeutic target for steatosis-induced liver damage, including NASH.