简介：neovascularization的形成是许多眼睛的脉管的疾病的一个普通病理学的特征，并且是在病人的视觉损失的一个重要原因。Neovascularization能引起网膜的出血，玻璃的出血，和另外的严肃的复杂并发症，导致视觉的损失。intraocularneovascularization的治疗是眼科学研究的焦点。在最近的年里，一些研究发现了autophagy是仔细与脉管的endothelial生长因素和neovascularization的形成有关。Autophagy被期望为intraocularneovascularization的治疗成为一个新目标。因此，这篇文章在autophagy和intraocularneovascularization的形成上考察研究。

简介：无

简介：无

简介：AbstractObjectives:Mechanistic target of rapamycin (mTOR) activation has been identified in keloid. This study aimed to identify the role of mTOR-dependent autophagy activity in keloid.Methods:We detected the expression of specific proteins representing mTOR activity and baseline autophagy levels in keloid tissues (KTs) and primary keloid fibroblasts (KFs) using immunohistochemical staining and western blotting. Simultaneously, the formation of acid vesicles was assessed by acridine orange staining in KFs. To investigate whether mTOR-dependent pathway mediated the regulation of autophagy machinery in keloid, we first validated whether mTOR inhibitors, rapamycin (100 nmol/L) and KU-0063794 (5 μmol/L), could inhibit mTOR activity in KFs by western blotting. Then we explored the reverse effects on autophagy activity induced by mTOR inhibitors in the presence of lysosomal protease inhibitors by western blotting.Results:It demonstrated elevated expression of mTOR, S6, and their activated forms in KTs, and an elevated expression of p-S6 Ser235/236 in KFs, suggesting mTOR was activated in keloid. Less LC3 and Beclin1 were expressed in the cytoplasm of KFs, whereas Ubiquitin was abundantly expressed in KTs compared with extra-lesional tissues. In addition, at the cellular level, an impeded conversion of LC3-I to LC3-II was shown in KFs and the formation of acid vesicles were also decreased in KFs compared with normal fibroblasts (NFs), indicating that autophagy activity is defective in keloid. mTOR inhibitors, Rapamycin (E-64d + pepstatin vs. rapamycin + E-64d + pepstatin: [0.88 ± 0.35] vs. [1.56 ± 0.46], F= 5.56, P= 0.049) and KU-0063794 (E-64d + pepstatin vs. KU-0063794 + E-64d + pepstatin: [0.92 ± 0.22] vs. [1.51 ± 0.25], F= 25.88, P= 0.011) can reverse the inhibition effect on autophagy of KFs while inhibiting mTOR activity.Conclusion:Autophagy machinery is inhibited in keloid which is regulated by mTOR-dependent pathway.

简介：AIM:Toinvestigatewhetherphotoreceptornecroptosisinducedbyz-VAD-FMK(pancaspaseinhibitor)wasinvolvedtheactivationofautophagyandwhetherNecrostatin-1,aspecificnecroptosisinhibitor,couldinhibitthisinductionofautophagyafterexperimentalretinaldetachment.METHODS:ExperimentalretinaldetachmentmodelswerecreatedinSprague-Dawleyratsbysubretinalinjectionofsodiumhyaluronateandsubretinalinjectionsofz-VAD-FMK,vehicleorz-VAD-FMKplusNecrostatin-1.Threedaysafterretinaldetachment,morphologicchangeswereobservedbytransmissionelectronmicroscopy.Inotheranimals,retinasweresubjectedtoimmunoprecipitationandWesternBlotting,thenprobedwithanti-RIP1,phosphoserine,LC-3IIorcaspase8antibody.RESULTS:Itwasprovedbyimmunoprecipitationandwesternblotting,thatphotoreceptornecroptosiswasmediatedbycaspase-8inhibitionandreceptorinteractingproteinkinase(RIP1)phosphorylationactivation.TransmissionelectronmicroscopeandwesternblottingresultsindicatedthatphotoreceptornecroptosiswasinvolvedtheLC-3IIandautophagosomesinduction.WealsodiscoveredNecrostatin-1couldinhibitRIP1phosphorylationandLC-3IIinduction.CONCLUSION:Thesedatafirstlyindicatephotoreceptornecroptosisisassociatedwiththeactivationofautophagy.Necrostatin-1protectsphotoreceptorsfromnecroptosisandautophagybydown-regulationofRIP1phosphorylationandLC-3II.

简介：AbstractAlternative splicing plays a pivotal role in the posttranscriptional regulation of gene expression, contributing to the generation of proteome diversity. Autophagy is a conserved cellular machinery governing degradation and recycling of long-lived or damaged proteins and organelles. However, there is limited knowledge of the roles of alternative splicing in autophagy, in particular mitochondrial selective autophagy, termed mitophagy. Emerging evidence suggests autophagy-related proteins (Atg), key molecules in autophagy process, are involved. This review highlights recent advances in the understanding of mechanisms by which alternative splicing affects the functions of ATG genes including BECN1, ATG5, ATG16L1, and Bim genes, and thus manipulates autophagy levels in various diseases. This review found that the effects of splicing of ATG genes generally result in inhibiting autophagy. However, very few of the many autophagy associated proteins have been studied. More research into the transcriptional and post-transcriptional regulation of splicing factors will be necessary to understand their roles in pathological conditions associated with autophagy and mitophagy.

简介：AbstractBackground:Sweat secreted by eccrine sweat glands is transported to the skin surface through the lumen. The eccrine sweat gland develops from the initial solid bud to the final gland structure with a lumen, but how the lumen is formed and the mechanism of lumen formation have not yet been fully elucidated. This study aimed to investigate the mechanism of lumen formation of eccrine gland organoids (EGOs).Methods:Human eccrine sweat glands were isolated from the skin for tissue culture, and the primary cultured cells were collected and cultured in Matrigel for 14 days in vitro. EGOs at different development days were collected for hematoxylin and eosin (H&E) staining to observe morphological changes and for immunofluorescence staining of proliferation marker Ki67, cellular motility marker filamentous actin (F-actin), and autophagy marker LC3B. Western blotting was used to detect the expression of Ki67, F-actin, and LC3B. Moreover, apoptosis was detected using a terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) apoptosis assay kit, and the expression of poly (ADP-ribose) polymerase and Caspase-3 was detected by Western blot. In addition, 3-methyladenine (3MA) was used as an autophagy inhibitor to detect whether the formation of sweat glands can be effectively inhibited.Results:The results showed that a single gland cell proliferated rapidly and formed EGOs on day 4. The earliest lumen formation was observed on day 6. From day 8 to day 14, the rate of lumen formation in EGOs increased significantly. The immunofluorescence and Western blot analyses showed that the expression of Ki67 gradually decreased with the increase in days, while the F-actin expression level did not change. Notably, the expression of autophagy marker LC3B was detected in the interior cells of EGOs as the apoptosis signal of EGOs was negative. Compared with the control group, the autophagy inhibitor 3MA can effectively limit the formation rate of the lumen and reduce the inner diameter of EGOs.Conclusion:Using our model of eccrine gland 3D-reconstruction in Matrigel, we determined that autophagy rather than apoptosis plays a role in the lumen formation of EGOs.

简介：

简介：AbstractCell death occurs in various tissues and organs in the body. It is a physiological or pathological process that has different effects. It is of great significance in maintaining the morphological function of cells and clearing abnormal cells. Pyroptosis, apoptosis, and necrosis are all modes of cell death that have been studied extensively by many experts and scholars, including studies on their effects on the liver, kidney, the heart, other organs, and even the whole body. The heart, as the most important organ of the body, should be a particular focus. This review summarizes the mechanisms underlying the various cell death modes and the relationship between the various mechanisms and heart diseases. The current research status for heart therapy is discussed from the perspective of pathogenesis.

简介：无

简介：RapamycintreatmenthasbeenshowntoincreaseautophagyactivityandactivateAktphosphorylation,suppressingapoptosisinseveralmodelsofischemiareperfusioninjury.However,littlehasbeenstudiedontheneuroprotectiveeffectsonspinalcordinjurybyactivatingAktphosphorylation.Wehypothesizedthatbotheffectsofrapamycin,theincreasedautophagyactivityandAktsignaling,wouldcontributetoitsneuroprotectiveproperties.Inthisstudy,acompressivespinalcordinjurymodelofratwascreatedbyananeurysmclipwitha30gclosingforce.Ratmodelswereintraperitoneallyinjectedwithrapamycin1mg/kg,followedbyautophagyinhibitor3-methyladenine2.5mg/kgandAktinhibitorIV1μg/kg.Westernblotassay,immunofluorescencestainingandterminaldeoxynucleotidyltransferase-mediateddUTPnickendlabelingassaywereusedtoobservetheexpressionofneuronalautophagymoleculeBeclin1,apoptosis-relatedmoleculesBcl-2,Bax,cytochromec,caspase-3andAktsignaling.OurresultsdemonstratedthatrapamycininhibitedtheexpressionofmTORininjuredspinalcordtissueandup-regulatedtheexpressionofBeclin1andphosphorylated-Akt.Rapamycinpreventedthedecreaseofbcl-2expressionininjuredspinalcordtissue,reducedBax,cytochromecandcaspase-3expressionlevelsandreducedthenumberofapoptoticneuronsininjuredspinalcordtissue24hoursafterspinalcordinjury.3-MethyladenineandAktinhibitorIVinterventionsuppressedtheexpressionofBeclin-1andphosphorylated-Aktininjuredspinalcordtissueandreducedtheprotectiveeffectofrapamycinonapoptoticneurons.TheaboveresultsindicatethattheneuroprotectiveeffectofrapamycinonspinalcordinjuryratscanbeachievedbyactivatingautophagyandtheAktsignalingpathway.

简介：

简介：真核细胞的房间的最惹人注目的形态特征是各种各样的围住膜的分隔空间的存在。包括细胞器和短暂运输中介，这些分隔空间不是静态的。更确切地说，蛋白质和膜的动态交换被需要维持细胞的动态平衡。膜动员的最戏剧的例子之一在macroautophagy的过程期间被看见。Macroautophagy是为长寿蛋白质和细胞器的降级的主要细胞的小径。响应环境暗示，例如饥饿或应力的另外的类型，房间生产唯一的膜结构，phagophore.Thephagophore扣押它形成一个双膜cytosolic泡的细胞质，anautophagosome。在结束之后，autophagosome与溶酶体熔化或在酵母的一个液泡，它交付水疗院放射激光与它的货物，和产生大分子一起毁坏内部autophagosome膜回来被释放进cytosol因为reuse.Autophagy因此是正在骑车过程，允许房间熬过滋养的限制的时期；然而，它有一个更宽的生理的角色，参予开发和老化，并且另外在对病原体侵略，癌症和某些neurodegenerative的保护疾病。在许多情况中，autophagy的角色通过autophagy相关的蛋白质的研究被识别，Atg6/Beclin1。这蛋白质是类脂化合物kinase建筑群的部分，并且最近的研究建议它在协调autophagy并且在反对apoptosis的细胞的死亡过程的thecytoprotective功能起一个中央作用。这里，我们总结我们Atg6/Beclin的当前的知识1在在细胞的不同模型有机体和它的唯一的功能。

简介：无

简介：Objective:TodeterminewhetherInterferon-alpha-2b(IFN-α2b)canmodulatetheautophagicresponseinhepatocellularcarcinomacells.Methods:HepatocellularcarcinomacellsweretreatedwithIFN-α2b.Autophagywasassessedbyacridineorangestaining,GFP-LC3dottedassay,transmissionelectronmicroscopyandimmunoblotting.Results:AcridineorangestainingshowedthatIFN-α2btriggeredtheaccumulationofacidicvesicularandautolysosomesinHepG2cells.TheacridineorangeHepG2cellratioswere(4.3±1.0)%,(6.9±1.4)%,and(13.1±2.3)%,respectively,aftertreatmentwith100,1,000,and10,000IU/mLIFN-α2bfor48h.AmarkedlypunctatepatternwasobservedinHepG2cellstreatedwith10,000IU/mLIFN-α2bfor48h,butonlydiffuseandweaklyfluorescentGFP-LC3punctawasobservedincontrolcells.HepG2cellstreatedwith10,000IU/mLIFN-α2bfor48hdevelopedautophagosome-likecharacteristics,includingsingle-ordouble-membranevacuolescontainingintactanddegradedcellulardebris.TheBeclin1andLC3-IIproteinexpressionwasup-regulatedbyIFN-α2btreatment.Conclusion:Autophagycanbeinducedinadose-dependentmannerbytreatmentwithIFN-α2binHepG2cells,andtheBeclin1signalingpathwaywasstimulatedbyIFN-α2b.

简介：AbstractBackground:Arteriosclerosis obliterans (ASO) is a major cause of adult limb loss worldwide. Autophagy of vascular endothelial cell (VEC) contributes to the ASO progression. However, the molecular mechanism that controls VEC autophagy remains unclear. In this study, we aimed to explore the role of the GRB2 associated binding protein 1 (GAB1) in regulating VEC autophagy.Methods:In vivo and in vitro studies were applied to determine the loss of adapt protein GAB1 in association with ASO progression. Histological GAB1 expression was measured in sclerotic vascular intima and normal vascular intima. Gain- and loss-of-function of GAB1 were applied in VEC to determine the effect and potential downstream signaling of GAB1.Results:The autophagy repressor p62 was significantly downregulated in ASO intima as compared to that in healthy donor (0.80 vs. 0.20, t= 6.43, P < 0.05). The expression level of GAB1 mRNA (1.00 vs. 0.24, t= 7.41, P < 0.05) and protein (0.72 vs. 0.21, t = 5.97, P < 0.05) was significantly decreased in ASO group as compared with the control group. Loss of GAB1 led to a remarkable decrease in LC3II (1.19 vs. 0.68, t = 5.99, P < 0.05), whereas overexpression of GAB1 significantly led to a decrease in LC3II level (0.41 vs. 0.93, t= 7.12, P < 0.05). Phosphorylation levels of JNK and p38 were significantly associated with gain- and loss-of-function of GAB1 protein.Conclusion:Loss of GAB1 promotes VEC autophagy which is associated with ASO. GAB1 and its downstream signaling might be potential therapeutic targets for ASO treatment.

简介：

简介：AbstractNon-alcoholic fatty liver disease (NAFLD) is a disorder of lipid metabolism. The lipotoxic intermediates of lipid metabolism cause mitochondrial dysfunction and endoplasmic reticulum stress. Organelle-specific autophagy is responsible for the removal of dysfunctional organelles to maintain intracellular homeostasis. Lipophagy contributes to lipid turnover by degrading lipid droplets. The level of autophagy changes during the course of NAFLD, and the activation of hepatocyte autophagy might represent a method of treating NAFLD.

简介：无

简介：AIM:Toinvestigatetheanti-tumoreffectsofnuclearfactor-κB(NF-κB)inhibitorSN50andrelatedmechanismsofSGC7901humangastriccarcinomacells.METHODS:MTTassaywasusedtodeterminethecytotoxiceffectsofSN50ingastriccancercelllineSGC7901.Hoechst33258stainingwasusedtodetectapoptosismorphologicalchangesafterSN50treatment.Activationofautophagywasmonitoredwithmonodansylcadaverine(MDC)stainingafterSN50treatment.Immunofluorescencestainingwasusedtodetecttheexpressionoflightchain3(LC3).MitochondrialmembranepotentialwasmeasuredusingthefluorescentprobeJC-1.Westernblottinganalysiswereusedtodeterminetheexpressionofproteinsinvolvedinapoptosisandautophagyincludingp53,p53upregulatedmodulatorofapoptosis(PUMA),damage-regulatedautophagymodulator(DRAM),LC3andBeclin1.Wedetectedtheeffectsofp53-mediatedautophagyactivationontheapoptosisofSGC7901cellswiththep53inhibitorpifithrin-α.RESULTS:TheviabilityofSGC7901cellswasinhibitedafterSN50treatment.Inductionsintheexpressionofapoptoticproteinp53andPUMAaswellasautophagicproteinDRAM,LC3andBeclin1weredetectedwithWesternblottinganalysis.SN50-treatedcellsexhibitedpunctuatemicrotubule-associatedprotein1LC3inimmunoreactivityandMDC-labeledvesiclesincreasedaftertreatmentofSN50byMDCstaining.CollapseofmitochondrialmembranepotentialΔψweredetectedfor6to24hafterSN50treatment.SN50-inducedincreasesinPUMA,DRAM,LC3andBeclin1andcelldeathwereblockedbythep53specificinhibitorpifithrin-α.CONCLUSION:Theanti-tumoractivityofNF-κBinhibitorsisassociatedwithp53-mediatedactivationofautophagy.