The effect of celastrol in combination with 5-fluorouracil on proliferation and apoptosis of gastric cancer cell lines

Background:Despite the availability of chemotherapy drugs such as 5-fluorouracil(5-FU),the treatment of some cancers such as gastric cancer remains challenging due to drug resistance and side effects.This study aimed to investigate the effect of celastrol in combination with the chemotherapy drug 5-FU on proliferation and induction of apoptosis in human gastric cancer cell lines(AGS and EPG85-257).Materials and Methods:In this in vitro study,AGS and EPG85-257 cells were treated with different concentrations of celastrol,5-FU,and their combination.Cell proliferation was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT)assay.The synergistic effect of 5-FU and celastrol was studied using Compusyn software.The DNA content at different phases of the cell cycle and apoptosis rate was measured usingflow cytometry.Results:Co-treatment with low concentrations(10%inhibitory concentration(IC10))of celastrol and 5-FU significantly reduced IC50(p<0.05)so that 48 h after treatment,IC50 was calculated at 3.77 and 6.9μM for celastrol,20.7 and 11.6μM for 5-FU,and 5.03 and 4.57μM for their combination for AGS and EPG85-257 cells,respectively.The mean percentage of apoptosis for AGS cells treated with celastrol,5-FU,and their combination was obtained 23.9,41.2,and 61.9,and for EPG85-257 cells 5.65,46.9,and 55.7,respectively.In addition,the 5-FU and celastrol-5-FU combination induced cell cycle arrest in the synthesis phase.Conclusions:Although celastrol could decrease the concentration of 5-fluorouracil that sufficed to suppress gastric cancer cells,additional studies are required to arrive at conclusive evidence on the anticancer effects of celastrol.

The Mechanism of Celastrol in the Treatment of Metastatic Lung Adenocarcinoma Revealed by Network Pharmacology and Molecular Docking

Background: Celastrol is an active ingredient extracted from Traditional Chinese Medicine (TCM), which can restrain the progression of lung cancer, whereas its underlying mechanism is unclear. In our study, the underlying mechanism of celastrol in the treatment of lung adenocarcinoma (LUAD) with metastasis was investigated by network pharmacology and molecular docking. Method: Potential targets of celastrol were collected from TCMSP, Batman-TCM and GeneCard database, and its potential targets were predicted using the STP platform and the TargetNet server. Metastasis marker genes (MGs) were obtained from the HCMDB. The genes correlated with LUAD were gathered from the GeneCard and OMIM database. And the common targets among celastrol potential targets, MGs and LUAD were analyzed. The protein-protein interaction (PPI) networks were obtained from the STRING database. SangerBox and the Xiantao bioinformatics tool were applied to visualize GO and KEGG analysis. Molecular docking tested the binding affinity between celastrol and core genes. Result: A total of 107 targets of celastrol against metastasis LUAD were obtained. The core targets were obtained from the PPI network, namely AKT1, JUN, MYC, STAT3, IL6, TNF, NFKB1, BCL2, IL1B, and HIF1A. GO and KEGG enrichment analysis indicated celastrol for the treatment of metastasis LUAD most refers to cellular response to chemical stress, DNA-binding transcription factor binding, transcription regulator complex and pathways in cancer. And some of these targets are associated with differential expressions and survival rates in LUAD. Moreover, Molecular docking shows celastrol can bind with BCL2 well by hydrogen bond and hydrophobic interaction. Conclusion: This finding roundly expounded the core genes and potential mechanisms of celastrol for the treatment of metastasis LUAD, offering the theoretical basis and antitumor mechanism of TCM in the treatment of lung cancer.

Celastrol promotes apoptosis of breast cancer MDA-MB-231 cells by targeting HSDL2

Objective:Celastrol is a pentacyclic triterpenoid extracted from the traditional Chinese medicinal herb,Tripterygium wilfordii.This study aims to provide a scientific basis for the rational development and use of celastrol in breast cancer.Method:A quantitative chemical biology approach was used to investigate the protein targets and molecular mechanisms of celastrol in breast cancer cells.Results:Low-concentration celastrol exerted an anti-tumor effect by directly binding to hydroxysteroid dehydrogenase-like 2(HSDL2)and inhibiting its expression.Moreover,the expression of the pro-apoptotic protein,Bcl-2-associated X(BaX),increased,the level of the anti-apoptotic protein,B-cell lymphoma-2(Bcl-2),decreased,and the rate of apoptosis increased.After the transfection of cells with si-HSDL2,the apoptosis rate was similar to that observed after the administration of celastrol.However,apoptosis was reversed by the overexpression of HSDL2.Furthermore,our mass spectrometry(MS)data indicated a relationship between HSDL2 and the mitogen-activated protein kinase(MAPK)signaling pathway.We also found that the expression of HSDL2 was directly related to the degree of extracellular signal-regulated kinase(ERK)phosphorylation.Conclusion:Celastrol may promote apoptosis by suppressing the HSDL2/MAPK/ERK signaling pathway.

Celastrol activates caspase-3/GSDME-dependent pyroptosis in tumor cells by inducing endoplasmic reticulum stress Author links open overlay panel

Objective:To investigate the pyroptosis-inducing effects of celastrol on tumor cells and to explore the potential mechanisms involved,specifically focusing on the role of the caspase-3/gasdermin E(GSDME)signaling pathway and the impact of endoplasmic reticulum(ER)stress and autophagy.Methods: Necrostatin-1(Nec-1),lactate dehydrogenase release(LDH)assay,and Hoechst/propidium iodide(PI)double staining were employed to validate the mode of cell death.Western blot was used to detect the cleavage of GSDME and the expression of light chain 3(LC3)and BIP.Results: Celastrol induced cell swelling with large bubbles,which is consistent with the pyroptotic phenotype.Moreover,treatment with celastrol induced GSDME cleavage,indicating the activation of GSDME-mediated pyroptosis.GSDME knockout via CRISPR/Cas9 blocked the pyroptotic morphology of celastrol in HeLa cells.In addition,cleavage of GSDME was attenuated by a specific caspase-3 inhibitor in celastrol-treated cells,suggesting that GSDME activation was induced by caspase-3.Mechanistically,celastrol induced endoplasmic reticulum(ER)stress and autophagy in HeLa cells,and other ER stress inducers produced effects consistent with those of celastrol.Conclusion: These findings suggest that celastrol triggers caspase-3/GSDME-dependent pyroptosis via activation of ER stress,which may shed light on the potential antitumor clinical applications of celastrol.

Celastrol inhibits inflammatory factors expression in glioblastoma

Background:Glioblastoma is one of the most common primary intracranial tumors of the central nervous system in adults.Although chemotherapy is an important component of glioblastoma treatment,its effectiveness remains unsatisfactory.Due to multiple immunosuppressive mechanisms,glioblastoma immunotherapy has not been effective in treating many patients as a result of the clinical breakthroughs in the field.Therefore,the development of cancer immunotherapy relies on the understanding of how tumors interact with the immune system and the analysis of their molecular determinants.This study identified the key interactions between immune cells in the glioma microenvironment using RNA microarrays and single-cell sequencing.Methods:First,we screened differentially expressed genes in tumor and control samples from GSE29796 and GSE50161 datasets using GEO2R.All differentially expressed genes were used to perform enrichment analysis and construct protein-protein interaction topological analysis to analyze the interaction between proteins.Using single-cell RNA sequencing data from the GSE162631 database,we identified immune cell types within the glioblastoma microenvironment,and validated the hub gene expression in these cells.In addition,based on the GEPIA and TIMER databases,hub genes were investigated and compared with immune infiltration to determine differential expression.Finally,CellChat was used to visualize the gene expression distribution and cell-to-cell communication analysis of the proteins between different types of cells.Results:We found that monocytes/macrophages may communicate with each other in the tumor microenvironment through MIF-(CD74+CXCR4)and MIF-(CD74+CD44).In addition,our study indicated that celastrol has the ability to inhibit inflammatory factors expression by MIF/CD74 signaling pathway in U87 cells.Conclusion:This study improved the effectiveness of cancer immunotherapy strategies and developed new ideas for immunotherapy that can be applied to glioblastoma.

基于CEL法熔滴冲击基板的TPF/FSI数值模拟

航空发动机和燃气轮机的高温工作环境会对其热端金属部件造成不可逆损伤,热障涂层(thermal barrier coatings,TBCs)能够承受高温和高压的侵蚀性环境,从而保证金属部件使用的安全性及可靠性。以空心Y_(2)O_(3)-ZrO_(2)(yttrium-stabilized zirconia,YSZ)粒子在等离子焰流中的2种形态,即全熔熔滴和空心熔滴为研究对象对其冲击铺展过程进行了模拟。基于ABAQUS/EXPLICIT耦合欧拉-拉格朗日(coupled Eulerian-Lagrangian,CEL)有限元方法,首先针对ABAQUS缺少相变模型,导致使用CEL方法计算熔滴铺展形貌失真的问题,给出了适用的动力黏度随温度变化的经验公式。此外,考虑周围空气对熔滴铺展过程的影响,提出了“两相流(two phase flow,TPF)/流-固耦合(fluid-structure-interaction,FSI)”2.5D模型,对熔滴冲击基板凝固成型及空气卷入的过程进行了模拟,并揭示了2种熔滴铺展形貌存在较大差异的机理,对制备隔热性能更优的热障涂层具有指导意义。

Celastrol targeting Nedd4 reduces Nrf2-mediated oxidative stress in astrocytes after ischemic stroke

Stroke is the second leading cause of death worldwide,and oxidative stress plays a crucial role.Celastrol exhibits strong antioxidant properties in several diseases;however,whether it can affect oxidation in cerebral ischemic-reperfusion injury(CIRI)remains unclear.This study aimed to determine whether celastrol could reduce oxidative damage during CIRI and to elucidate the underlying mechanisms.Here,we found that celastrol attenuated oxidative injury in CIRI by upregulating nuclear factor E2-related factor 2(Nrf2).Using alkynyl-tagged celastrol and liquid chromatography-tandem mass spectrometry,we showed that celastrol directly bound to neuronally expressed developmentally downregulated 4(Nedd4)and then released Nrf2 from Nedd4 in astrocytes.Nedd4 promoted the degradation of Nrf2 through K48-linked ubiquitination and thus contributed to astrocytic reactive oxygen species production in CIRI,which was significantly blocked by celastrol.Furthermore,by inhibiting oxidative stress and astrocyte activation,celastrol effectively rescued neurons from axon damage and apoptosis.Our study uncovered Nedd4 as a direct target of celastrol,and that celastrol exerts an antioxidative effect on astrocytes by inhibiting the interaction between Nedd4 and Nrf2 and reducing Nrf2 degradation in CIRI.

Mechanistic engineering of celastrol liposomes induces ferroptosis and apoptosis by directly targeting VDAC2 in hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is one of most common and deadliest malignancies.Celastrol(Cel),a natural product derived from the Tripterygium wilfordii plant,has been extensively researched for its potential effectiveness in fighting cancer.However,its clinical application has been hindered by the unclear mechanism of action.Here,we used chemical proteomics to identify the direct targets of Cel and enhanced its targetability and antitumor capacity by developing a Cel-based liposomes in HCC.We demonstrated that Cel selectively targets the voltage-dependent anion channel 2(VDAC2).Cel directly binds to the cysteine residues of VDAC2,and induces cytochrome C release via dysregulating VDAC2-mediated mitochondrial permeability transition pore(mPTP)function.We further found that Cel induces ROS-mediated ferroptosis and apoptosis in HCC cells.Moreover,coencapsulation of Cel into alkyl glucoside-modified liposomes(AGCL)improved its antitumor efficacy and minimized its side effects.AGCL has been shown to effectively suppress the proliferation of tumor cells.In a xenograft nude mice experiment,AGCL significantly inhibited tumor growth and promoted apoptosis.Our findings reveal that Cel directly targets VDAC2 to induce mitochondria-dependent cell death,while the Cel liposomes enhance its targetability and reduces side effects.Overall,Cel shows promise as a therapeutic agent for HCC.

基于CEL方法的瓷砖智能化铺贴质量数值分析

基于耦合的欧拉-拉格朗日方法(CEL方法)建立瓷砖-瓷砖胶-混凝土流固耦合施工模型,模拟研究机器人的布料方式、布料管数和荷载类型对瓷砖回弹量、接触面积、缺陷分布等铺贴质量指标的影响规律和机理。研究结果表明:与垂直布料相比,水平布料瓷砖胶的塑性变形较小,胶条形态保持稳定且呈等间距分布。布料管数增加对瓷砖回弹量与接触面积均无影响,但会使瓷砖-瓷砖胶的界面缺陷明显变窄且分布更加均匀。相较于单一的直压荷载,耦合垂直振动荷载引起的不均匀塑性变形导致瓷砖回弹量和瓷砖-瓷砖胶接触面积同时降低;而耦合剪切振动荷载既有利于消除瓷砖回弹量又能显著增大瓷砖-瓷砖胶接触面积,有效减少界面缺陷。

Celastrol mitigates inflammation in sepsis by inhibiting the PKM2-dependent Warburg effect

Background: Sepsis involves life-threatening organ dysfunction and is caused by a dysregulated host response to infection. No specific therapies against sepsis have been reported. Celastrol(Cel) is a natural anti-inflammatory compound that shows potential against systemic inflammatory diseases. This study aimed to investigate the pharmacological activity and molecular mechanism of Cel in models of endotoxemia and sepsis.Methods: We evaluated the anti-inflammatory efficacy of Cel against endotoxemia and sepsis in mice and macrophage cultures treated with lipopolysaccharide(LPS). We screened for potential protein targets of Cel using activity-based protein profiling(ABPP). Potential targets were validated using biophysical methods such as cellular thermal shift assays(CETSA) and surface plasmon resonance(SPR). Residues involved in Cel binding to target proteins were identified through point mutagenesis, and the functional effects of such binding were explored through gene knockdown.Results: Cel protected mice from lethal endotoxemia and improved their survival with sepsis, and it significantly decreased the levels of pro-inflammatory cytokines in mice and macrophages treated with LPS(P <0.05). Cel bound to Cys424 of pyruvate kinase M2(PKM2), inhibiting the enzyme and thereby suppressing aerobic glycolysis(Warburg effect). Cel also bound to Cys106 in high mobility group box 1(HMGB1) protein, reducing the secretion of inflammatory cytokine interleukin(IL)-1β. Cel bound to the Cys residues in lactate dehydrogenase A(LDHA).Conclusions: Cel inhibits inflammation and the Warburg effect in sepsis via targeting PKM2 and HMGB1 protein.

基于CEL法的破损边界附近气泡动力学特性研究

为研究船舶受到水下爆炸冲击波打击后遭受气泡二次冲击,对船体产生破损甚至折断这一类问题,气泡与破损边界的相互作用机理。将破舱壁面简化为具有圆形破口的弹性壁面,基于耦合欧拉-拉格朗日法建立了破损边界附近水下爆炸气泡动力学模型,将数值结果与模型试验结果进行对比,验证了所建立方法的有效性。在此基础上,研究了气泡射流、气泡体积、脉动周期,以及气泡对壁面冲击损伤的影响,探明了不同破口参数下气泡的脉动规律和射流穿透破口、撕裂等特征规律。为高能武器的研发和舰船破损后安全防护提供理论和技术支撑。

湖羊CEL基因多态性及其与眼肌面积性状的关联分析

旨在探讨羧基酯脂肪酶基因(CEL)的单核苷酸多态性与湖羊眼肌面积的相关性。选取1049只具有准确表型记录且健康的湖羊,屠宰后测定其眼肌面积,采集血液提取基因组DNA,通过PCR和KASPar SNP分型技术对CEL基因进行目的片段扩增和基因分型,并与湖羊眼肌面积进行关联分析,采用RT-qPCR检测CEL基因在湖羊10种组织中的表达情况。结果显示,湖羊CEL基因在不同组织中均有表达,且在十二指肠中表达量最高;该基因存在一个g.4039718 C>T的同义突变,呈中等多态。性状关联分析表明,该多态位点与湖羊的眼肌面积显著关联,TT型个体的眼肌面积显著高于CC型个体。描述性统计结果显示,眼肌面积的变异系数为12.29%,具有较高的选择潜力。相关性分析表明,眼肌面积与体质量、体高、体长、胸围、屠宰率、胴体质量和宰前活质量等生长性状和屠宰性状呈正相关。结果表明,g.4039718 C>T多态位点可作为候选分子标记用于湖羊眼肌面积性状的遗传改良。

Study of molecular mechanisms underlying the medicinal plant Tripterygium wilfordii-derived compound celastrol in treating diabetic nephropathy based on network pharmacology and molecular docking

Background:Diabetic nephropathy(DN)is a serious complication of diabetes with rising prevalence worldwide.We aimed to explore the anti-DN mechanisms of the compound celastrol derived from the medicinal plant Tripterygium wilfordii.Methods:Celastrol-related targets were obtained from Herbal Ingredients’Targets(HIT)and GeneCards databases.DN-related targets were retrieved from GeneCards,DisGeNET,and Therapeutic Targets Database(TTD).A Protein-protein interaction(PPI)network was established using the Search Tool for the Retrieval of Interacting Genes(STRING)database.Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed using ClusterProfiler.The cytoHubba plugin was used to select the top 10 hub targets.Molecular docking was performed employing PyMOL and AutoDock software.Cell counting kit-8(CCK-8)and flow cytometry assays were used to detect the viability and apoptosis of NRK-52E cells,respectively.The mRNA expression levels of mitogen-activated protein kinase 3(MAPK3),tumor necrosis factor(TNF),and AKT serine/threonine kinase 1(AKT1)in NRK-52E cells were assessed using quantitative real-time polymerase chain reaction(qRT-PCR).Results:We obtained sixty-six overlapping targets of celastrol and DN.GO and KEGG analyses demonstrated that the core targets of celastrol and DN were mainly involved in the inflammatory and immune response,oxidative stress,advanced glycation end products(AGEs)and their receptors(RAGEs)(AGE-RAGE),TNF,interleukin 17(IL-17),and MAPK signaling pathways.Finally,based on the good binding activity with celastrol,MAPK3,TNF,and AKT1 were identified as the foremost targets of celastrol.We observed that celastrol enhanced the viability of high glucose(HG)-treated NRK-52E cells and inhibited apoptosis in the in vitro assays.Moreover,celastrol decreased the mRNA expression levels of MAPK3,TNF,and AKT1 in high glucose(HG)-treated NRK-52E cells.Conclusion:Celastrol may treat DN by targeting APK3,TNF,and AKT1 and regulating inflammatory responses and oxidative stress through the AGE-RAGE,TNF,IL-17,and MAPK signaling pathways.

基于CEL算法的神狐海域泥火山内部物质运动过程数值模拟

以南海北部神狐海域地震剖面识别的泥火山为基础,基于耦合的欧拉-拉格朗日算法(CEL)模拟了泥火山内部物质的运动过程。通过不同阶段的对比分析,研究探讨了泥火山的形成机理,总结了泥火山内部物质的运动特征及规律。数值模型的结果显示泥火山形成过程中物质的运移具有明显的阶段性变化,展现了物质在运移时的应力与应变的不规则变化规律,本文根据这些变化分析了泥火山形成过程中应力、应变与能量的强度变化趋势。地层在物质向上刺穿过程中受力分布表现出明显的不均匀性,中间部位的突起破裂引起两侧地层的隆升。由模拟结果提供的泥火山形成过程中内部物质的特征,可以推动对泥火山形成机制及过程的进一步认识,同时也能对深部底辟构造以及油气迁移提供理论指导。

基于CEL方法对LRMP测试浅层软土强度离心试验的模拟

软土海床/河床浅层土体强度对管线铺设等工程设计具有重要影响。基于CEL方法对LRMP测试浅层软土强度离心试验进行模拟,验证了CEL方法的可靠性,讨论了土体弹性模量和LRMP入土深度对水平受力系数的影响。计算结果显示,土体弹性模量对CEL计算得到的水平受力系数影响不大,可根据模拟土体的变形特性在300 s_(u)~900 s_(u)选取弹性模量数值;使用LRMP测量土体0.4 D~1 D强度时,可取水平受力系数为10.5,但土体强度梯度对水平受力系数有显著影响,当测量自重固结土时,使用N_(h)=10.5会低估土体的真实强度,需通过标定试验确定目标土体N h随土体强度梯度的变化规律,计算得到的规律可为测量软土浅层平均强度的相关试验设计和数值模拟提供技术参考。

曲线顶管底幕法施工对沉船扰动的CEL数值模拟

依托“长江口二号”沉船打捞工程,采用耦合欧拉-拉格朗日法对极小曲率半径矩形曲线顶管底幕法沉船打捞工程进行数值模拟分析,获得底幕施工过程中顶推力以及船体竖向位移的动态变化,并与模型试验结果进行了对比验证.结果表明:所提的数值模型模拟结果和模型试验结果吻合较好,能较精确地预测沉船的变位和顶管的顶推力变化;最后一根管节推进为最危险工况,需要加强施工控制;采用左右对称推进而非顺序推进能够有效降低施工对沉船和土体的扰动.

Celastrol inhibits migration, proliferation and transforming growth factor-β2-induced epithelial-mesenchymal transition in lens epithelial cells

AIM: To investigate the mechanism of celastrol in inhibiting lens epithelial cells(LECs) fibrosis, which is the pathological basis of cataract.METHODS: Human LEC line SRA01/04 was treated with celastrol and transforming growth factor-β2(TGF-β2). Wound-healing assay, proliferation assay, flow cytometry, real-time polymerase chain reaction(PCR), Western blot and immunocytochemical staining were used to detect the pathological changes of celastrol on LECs. Then, we cultured Sprague-Dawley rat lens in medium as a semi-in vivo model to find the function of celastrol further.RESULTS: We found that celastrol inhibited the migration of LECs, as well as proliferation(P<0.05). In addition, it induced the G2/M phase arrest by cell cyclerelated proteins(P<0.01). Moreover, celastrol inhibited epithelial-mesenchymal transition(EMT) by the blockade of TGF-β/Smad and Jagged/Notch signaling pathways.CONCLUSION: Our study demonstrates that celastrol could inhibit TGF-β2-induced lens fibrosis and raises the possibility that celastrol could be a potential novel drug in prevention and treatment of fibrotic cataract.

CEL法与ALE法在海底管土作用数值仿真中的应用

针对海底管道与海床相互作用有限元仿真中普通拉格朗日单元因大变形而畸变的问题,需要选用更合适的仿真方法。本文首先分别采用针对网格大变形问题发展出来的任意拉格朗日欧拉(ALE)法以及欧拉-拉格朗日耦合(CEL)法建立有限元模型,借助模型试验数据验证数值仿真方法的合理性,并通过网格无关性分析,确定了ALE法与CEL法适合的单元尺寸。然后分别利用拉格朗日法、ALE法以及CEL法计算了海底管道垂向与侧向土体反力,通过对比分析得出各种方法在研究海底管土作用时的优、缺点。最后进行了管土作用试验,发现CEL法能够较好地反映管土作用中的速度放大效应。

Celastrol Induces Apoptosis and Autophagy via the AKT/mTOR Signaling Pathway in the Pituitary ACTH-secreting Adenoma Cells

Objective Pituitary adrenocorticotropic hormone(ACTH)-secreting adenoma is a relatively intractable endocrine adenoma that can cause a range of severe metabolic disorders and pathological changes involving multiple systems.Previous studies have shown that celastrol has antitumor effects on a variety of tumor cells via the AKT/mTOR signaling.However,whether celastrol has pronounced antitumor effects on pituitary ACTH-secreting adenoma is unclear.This study aimed to identify a new effective therapeutic drug for pituitary ACTH-secreting adenoma.Methods Mouse pituitary ACTH-secreting adenoma cells(AtT20 cells)were used as an experimental model in vitro and to establish a xenograft tumor model in mice.Cells and animals were administered doses of celastrol at various levels.The effects of celastrol on cell viability,migration,apoptosis and autophagy were then examined.Finally,the potential involvement of AKT/mTOR signaling in celastrol’s mechanism was assessed.Results Celastrol inhibited the proliferation and migration of pituitary adenoma cells in a time-and concentration-dependent manner.It blocked AtT20 cells in the G0/G1 phase,and induced apoptosis and autophagy by downregulating the AKT/mTOR signaling pathway.Similar results were obtained in mice.Conclusion Celastrol exerts potent antitumor effects on ACTH-secreting adenoma by downregulating the AKT/mTOR signaling in vitro and in vivo.

Celastrol Protects TGF-β1-induced Endothelial-mesenchymal Transition

The endothelial-to-mesenchymal transition（End MT） in endothelial cells contributes to the development of cardiac fibrosis,ultimately leading to cardiac remodeling.In this study,the effects and molecular mechanisms of celastrol（CEL） on transforming growth factor-β1（TGF-β1）-induced End MT in human umbilical vein endothelial（HUVEC-12） cells were investigated.The presented data demonstrated that CEL significantly blocked the morphology change of HUVEC-12 cells induced by TGF-β1 without cell cytotoxicity.In accordance with these findings,CEL blocked TGF-β1-induced EndM T as evidenced by the inhibition of the mesenchymal markers,including collagen Ⅰ,Ⅲ,α-SMA,fibronectin m RNA expression,and the increase in the m RNA expression of endothelial cell marker CD31.These changes were also confirmed by double immunofluorescence staining of CD31 and vimentin.The in vitro scratch assay showed that CEL inhibited the migration capacity of the transitioned endothelial cells induced by TGF-β1.Further experiments showed that the beneficial effect of CEL on blocking the End MT in HUVEC-12 cells was associated with the suppression of the TGF-β1/Smads signalling pathway,which was also confirmed by the inhibition of its downstream transcription factor snail1,twist1,twist2,ZEB1 and ZEB2.These results indicate that CEL blocks TGF-β1-induced End MT through TGF-β1/Smads signalling pathway and suggest that it may be a feasible therapy for cardiac fibrosis diseases.