Morin is a functional flavonoid commonly found in human diet.Compared to being used solely,it is evident that morin can be more effective as a drug adjuvant.However,research on the combined effect and its correspondin...Morin is a functional flavonoid commonly found in human diet.Compared to being used solely,it is evident that morin can be more effective as a drug adjuvant.However,research on the combined effect and its corresponding mechanism is limited.Here,we found that morin significantly potentiated the inhibitory effects of the natural compound celastrol on the proliferation of lung cancer cells.Morin and celastrol synergistically exhibit marked apoptosis induction in lung cancer cells,accompanied by changes in the abundance of apoptosis-related proteins.Transcriptome analyses revealed that the combination of morin and celastrol had a significant impact on the number of differentially expressed genes in lung cancer cells.Among these genes,BIRC3 was one of the most significantly different ones,which plays a crucial role in the process of tumor resistance to apoptosis.In addition,several genes identified are primarily associated with intracellular signal transduction pathways,specifically the NF-κB signaling pathway.Importantly,the treatment combining morin and celastrol in tumor-bearing mice results in a synergistic effect that significantly suppressed tumor growth.These findings indicate that morin could be a promising functional adjuvant,and the combination of morin and celastrol has potential for the treating lung cancer.展开更多
Combining cytotoxic drugs with tumor microenvironment(TME)modulator agents is an effective strategy to enhance anti-tumor effects.In this study,two natural anti-tumor active ingredients celastrol(CEL)and glycyrrhetini...Combining cytotoxic drugs with tumor microenvironment(TME)modulator agents is an effective strategy to enhance anti-tumor effects.In this study,two natural anti-tumor active ingredients celastrol(CEL)and glycyrrhetinic acid(GA)were combined for tumor treatment.In order to ensure the precise co-delivery and controllable synchronous release of combined drugs to tumors,it is necessary to construct a suitable nano-drug delivery platform.Based on this,we coupled hyaluronic acid(HA)with CEL by amide reaction to obtain an amphiphilic polymer prodrug HA-SS-CEL,and GA was spontaneously loaded into polymer micelles by self-assembly to obtain G/HSSC-M.G/HSSC-M has ideal size distribution,redox-responsive synchronous drug release,enhanced tumor cell internalization and in vivo tumor targeting.Compared with free drugs,the construction of multifunctional polymer micelles makes G/HSSC-M show better anticancer effect at the same concentration,and can significantly inhibit the proliferation and migration of HepG2 and 4T1 cells.In the in vivo experiments,G/HSSC-M achieved a tumor inhibition rate as high as 75.12%in H22 tumor-bearing mice.The mechanism included regulation of M1/M2 macrophage polarization,inhibition of Janus kinase 1/signal transducer and activator of transcription 3(JAK1/STAT3)signaling pathway,and remodeling of tumor blood vessels.Therefore,the development of prodrug micelles coloaded with CEL and GA provides a promising drug co-delivery strategy for combined cancer therapy.展开更多
Objective:To investigate the mechanisms underlying the renoprotective effects of celastrol,a bioactive compound extracted from the traditional Chinese medicinal plant Tripterygium wilfordii in diabetic kidney disease(...Objective:To investigate the mechanisms underlying the renoprotective effects of celastrol,a bioactive compound extracted from the traditional Chinese medicinal plant Tripterygium wilfordii in diabetic kidney disease(DKD).Methods:We established a DKD model using db/db mice and investigated the protective mechanisms of celastrol against DKD progression using integrated analysis of 16S rRNA sequencing and transcriptome analysis.We evaluated colon tissue damage using hematoxylin and eosin and immunofluorescence staining.In addition,16S rRNA sequencing and transcriptomic analyses were performed to explore the potential mechanisms of celastrol.Immunofluorescence staining,Western blotting and real-time quantitative polymerase chain reaction analysis were performed to confirm the PPAR signaling pathway related proteins in kidney tissues.Results:Celastrol alleviated colon injury and increased the expression of mucosal barrier markers,particularly occludin and zonula occludens-1.The 16S rRNA gene sequencing analysis demonstrated that treatment with celastrol altered the diversity and abundance of the gut microbiota.Spearman’s correlation analysis further revealed significant associations among gut microbial,renal injury markers,and serum lipid profiles.A subsequent renal transcriptome analysis revealed that celastrol significantly modulated the renal transcriptional landscape,primarily by regulating genes associated with the PPAR signaling pathway and lipid metabolism.Further investigations demonstrated that celastrol significantly downregulated adipose differentiation-related protein expression and attenuated DKD progression by activating the PPAR pathway.Conclusions:This study demonstrates that celastrol alleviates both colonic and renal injuries by modulating the gut-kidney axis through PPAR-mediated lipid metabolism regulation,indicating its potential as a therapeutic approach for DKD management.展开更多
BACKGROUND The role of NLR family pyrin domain containing 3(NLRP3)in post-endoscopic submucosal dissection(ESD)esophageal stricture remains incompletely understood.The effect of celastrol(CEL)on the prevention of esop...BACKGROUND The role of NLR family pyrin domain containing 3(NLRP3)in post-endoscopic submucosal dissection(ESD)esophageal stricture remains incompletely understood.The effect of celastrol(CEL)on the prevention of esophageal strictures has not yet been investigated.AIM To explore the effect of CEL on the prevention of esophageal stricture in rats.METHODS NLRP3,interleukin(IL)-1β,and IL-18 mRNA levels were measured in patients’tissues after esophageal ESD.NLRP3 expression in esophageal fibroblasts was determined using immunohistochemistry and immunofluorescence staining.Lentiviral transfection was used to induce NLRP3 overexpression and thioredoxin reductase 1(TXNRD1)silencing.The CCK8 assay was used to determine the optimal CEL concentration.Reactive oxygen species(ROS)generation was detected via fluorescence and flow cytometry.Masson’s trichrome staining and barium esophagography were performed to assess collagen deposition and esophageal stenosis.RESULTS The mRNA levels of NLRP3 and IL-1βwere higher in human tissues from the ESD resection bed than in normal esophageal mucosa.NLRP3 overexpression in primary rat esophageal fibroblasts led to high collagen 1 expression.Thus,NLRP3 participated in esophageal inflammation and tissue repair after ESD.Comparable to prednisolone,CEL significantly inhibited NLRP3 activation in vitro and in vivo,and esophageal strictures were markedly alleviated.Mechanistically,CEL upregulated TXNRD1 expression and reduced ROS production,thereby inhibiting NLRP3 expression.This effect was reversed by TXNRD1 silencing.Furthermore,TXNRD1 interacted with NLRP3 and promoted its ubiquitination.CONCLUSION CEL is a promising alternative therapeutic agent for the prevention of post-ESD esophageal strictures.展开更多
OBJECTIVE： Celastrol has been established as a nuclear factor-κB（NF-κB） activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted ...OBJECTIVE： Celastrol has been established as a nuclear factor-κB（NF-κB） activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted that int erleukin-1 receptor-associated kinases（IRA Ks） are potential celastrol targets, and hypothesized that targeting IRAKs might be one way that celastrol inhibits NF-κB. This is because IRAKs are key molecules for some crucial pathways to activate NF-κB（e.g., the inter leukin-1 receptor（IL-1R）/Toll- like receptor（TLR） superfamily）.METHODS： The human hepatocellular cell line（Hep G2） treated with palmitic acid（PA） was used as a model for stimulating TLR4/NF-κB activation, in order to observe the potential effects of celastrol in IRAK regulation and NF-κB inhibition. The transfection of small interfering RNA was used for down-regulating TLR4, IRAK1 and IRAK4, and the Western blot method was used to detect changes in the protein expressions.RESULTS： The results showed that celastrol could effectively inhibit PA-caused TLR4-dependent NF-κB activation in the Hep G 2 cells; PA also activated IRAKs, which were inhibited by celastrol. Knocking down IRAKs abolished PA-caused NF-κB activation.CONCLUSION： The results for the first time show that targeting IRAKs is one way in which celastrol inhibits NF-κB activation.展开更多
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 transformi...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.展开更多
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 effectiven...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.展开更多
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 ...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.展开更多
Excessive oxidative stress impairs cartilage matrix metabolism balance,significantly contributing to osteoarthritis(OA)development.Celastrol(CSL),a drug derived from Tripterygium wilfordii,has recognized applications ...Excessive oxidative stress impairs cartilage matrix metabolism balance,significantly contributing to osteoarthritis(OA)development.Celastrol(CSL),a drug derived from Tripterygium wilfordii,has recognized applications in the treatment of cancer and immune system disorders,yet its antioxidative stress mechanisms in OA remain underexplored.This study aimed to substantiate CSL’s chondroprotective effects and unravel its underlying mechanisms.We investigated CSL’s impact on chondrocytes under both normal and inflammatory conditions.In vitro,CSL mitigated interleukin(IL)-1β-induced activation of proteinases and promoted cartilage extracellular matrix(ECM)synthesis.In vivo,intra-articular injection of CSL ameliorated cartilage degeneration and mitigated subchondral bone lesions in OA mice.Mechanistically,it was found that inhibiting nuclear factor erythroid 2-related factor 2(NRF2)abrogated CSL-mediated antioxidative functions and exacerbated the progression of OA.This study is the first to elucidate the role of CSL in the treatment of OA through the activation of NRF2,offering a novel therapeutic avenue for arthritis therapy.展开更多
Inflammation is a defense mechanism associated with a wide range of diseases.Celastrol is a small molecule isolated from traditional Chinese medicine with potent anti-inflammation activity.In this study,we established...Inflammation is a defense mechanism associated with a wide range of diseases.Celastrol is a small molecule isolated from traditional Chinese medicine with potent anti-inflammation activity.In this study,we established an integrated quantitative proteomics strategy to investigate the acute response to celastrol treatment in a rat macrophage cell line challenged with lipopolysaccharide(LPS).Both stableisotopic based non-targeted quantitative profiling and PRM-based targeted quantitation methods were employed.Dimethyl-labeling based non-targeted profiling revealed 28 and 52 proteins that significantly up-and down-regulated by celastrol.Bioinformatics analysis pinpoint key signaling pathways affected.Seven proteins were selected for examining their time-dependent regulatory pattern in response to celastrol using targeted PRM quantitation.The abundance of mRNA at multiple time-points of selected proteins was also examined.Celastrol induced an acute response of selected key transcriptional factors in terms of mRNA or protein abundance within one hour.Interestingly,regulatory trend of mRNA and protein abundance suggested a novel dual mechanism of celastrol in the terms of acute antiinflammation.The integrated quantitative proteomic strategy established in this study constitutes an efficient workflow to characterize key components and their time-dependent regulatory pattern for monitoring drug response.展开更多
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 sy...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.展开更多
OBJECTIVE: To investigate the efficacy of celastrol treatment of hepatocellular carcinoma(HCC) cells in vitro and in vivo and to propose a mechanism of action.METHODS: A human Hep G2 liver cancer cell line and a xenog...OBJECTIVE: To investigate the efficacy of celastrol treatment of hepatocellular carcinoma(HCC) cells in vitro and in vivo and to propose a mechanism of action.METHODS: A human Hep G2 liver cancer cell line and a xenograft tumor model were used to investigate the effects of celastrol on HCC in vitro and in vivo. A CCK-8 kit was used to detect cell viability.Flow cytometry and terminal-deoxynucleoitidyl transferase mediated nick end labeling staining were used to detect apoptosis. Western blotting and immunohistochemistry were used to detect the expression of cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, cleaved-PARP, mammalian target of rapamycin(mTOR), and p-mTOR. Hematoxylin-eosin staining was used to observe the tissue morphology.RESULTS: Celastrol decreased the viability of Hep G2 cells and induced apoptosis. Western blot assays indicated that celastrol up-regulated cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, and cleaved-PARP by inhibiting the phosphorylation of mTOR in Hep G2 cells. Moreover,celastrol inhibited the tumor growth in a xenograft model. Celastrol also induced caspase-dependent apoptosis(up-regulation of cleaved-caspase-3,-8,-9, and cleaved-PARP) and inhibited the activation of mTOR in vivo.CONCLUSION: Celastrol induces caspase-dependent apoptosis in HCC cells by inhibiting the activation of mTOR.展开更多
Celastrol,a Chinese herbal medicine,has exhibited anticancer activity in many types of cancer cells.However,the further clinical application of celastrol is restricted by its poor water solubility and serious side eff...Celastrol,a Chinese herbal medicine,has exhibited anticancer activity in many types of cancer cells.However,the further clinical application of celastrol is restricted by its poor water solubility and serious side effects.Furthermore,the apoptosis mechanism of tumor cells induced by celastrol has not been exhausted yet.In this study,we developed a reduction sensitive polymeric vector for tumor-targeted celastrol delivery.And our researches indicated that the celastrol could be delivered by reductionsensitive nanomedicine(RSNMs)with a controlled release strategy.Meanwhile,the cell uptake results indicated that excellent reduction-sensitive behavior of RSNMs could effectively accelerate celastrol into the human retinoblastoma(RB)cell.The cell cytotoxicity assay demonstrated that celastrol inhibited proliferation of human RB Y79 cells growth in a dose-dependent manner.Furthermore,the results of flow cytometry and terminal dUTP nick-end labeling(TUNEL)staining showed that celastrol induced apoptosis of the RB Y79 cells,and revealed a time-dependent increase in apoptosis induction of RB Y79 cells.The results of western blotting showed that celastrol induced the apoptosis of human RB Y79 cells involving the activation of caspase-3 and caspase-9.In conclusion,our results revealed that RSNMs may be utilized as a novel therapy for retinoblastoma.展开更多
Tripterygium hypoglaucum(Levl.)Hutch,a traditional Chinese medicinal herb with a long history of use,is widely distributed in China.One of its main active components,celastrol,has great potential to be developed into ...Tripterygium hypoglaucum(Levl.)Hutch,a traditional Chinese medicinal herb with a long history of use,is widely distributed in China.One of its main active components,celastrol,has great potential to be developed into anti-cancer and anti-obesity drugs.Although it exhibits strong pharmacological activities,there is a lack of sustainable sources of celastrol and its derivatives,making it crucial to develop novel sources of these drugs through synthetic biology.The key step in the biosynthesis of celastrol is considered to be the cyclization of 2,3-oxidosqualene into friedelin under the catalysis of 2,3-oxidosqualene cyclases.Friedelin was speculated to be oxidized into celastrol by cytochrome P450 oxidases(CYP450s).Here,we reported a cytochrome P450 ThCYP712K1 from Tripterygium hypoglaucum(Levl.)Hutch that catalyzed the oxidation of friedelin into polpuonic acid when heterologously expressed in yeast.Through substrate supplementation and in vitro enzyme analysis,ThCYP712K1 was further proven to catalyze the oxidation of friedelin at the C-29 position to produce polpunonic acid,which is considered a vital step in the biosynthesis of celastrol,and will lay a foundation for further analysis of its biosynthetic pathway.展开更多
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 t...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.展开更多
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-infl...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.展开更多
AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was...AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was treated with celastrol to evaluate this natural compound's impact on CNV.Western blot was applied to explore the possible mechanism for it.For in vitro assay,triplicate for each group was repeated at least three times.For in vivo assay,each group contains 5 mice.RESULTS:Celastrol supressed tube formation and aortic ring sprout neovascularization.In vitro assay exhibited that celastrol inhibiting vascular endothelial growth factor(VEGF)-induced proliferation and migration of human umbilical vein endothelial cells and human choroidal endothelial cells,and by blocking VEGF signaling.Furthermore,intraperitoneal administration of celastrol significantly reduced the area of laser-induced CNV in an in vivo mouse model.By day 14,the area of CNV had decreased by 49.15%and 80.26%in the 0.1 mg/kg celastrol-treated group(n=5)and in the 0.5 mg/kg celastrol treated group(n=5),respectively,compared to the vehicle-treated group(n=5).CONCLUSION:Celastrol inhibits CNV by inhibiting VEGF-induced proliferation and migration of vascular endothelial cells,indicating that celastrol is a potent,natural therapeutic compound for the prevention of CNV.展开更多
OBJECTIVE: To investigate the in-depth pharmacological mechanisms of celastrol in children neuroblastoma treatment. METHODS: In the current study, we examined the effects of celastrol on children neuroblastoma cells v...OBJECTIVE: To investigate the in-depth pharmacological mechanisms of celastrol in children neuroblastoma treatment. METHODS: In the current study, we examined the effects of celastrol on children neuroblastoma cells viability and proliferation by cell counting kit-8 assay and colony formation assay. Annexin V-FTIC and PI staining were applied to determine cell apoptosis after celastrol treatment. ROS generation levels were examined by 2′, 7′-dichloroflfluorescin diacetate. RESULTS: We found that celastrol could suppress the proliferation of children neuroblastoma cells with few effects on normal cell lines in vitro. Further mechanisms studies have shown that celastrol inhibited cell cycle progression and induced cell apoptosis in QDDQ-NM and SH-SY5Y cells. In addition, ROS production might involve in celastrol-mediated apoptotic cell death in children neuroblastoma cells by activating caspase death pathway. CONCLUSIONS: Our findings demonstrated that celastrol could promote ROS generation-induced apoptosis in neuroblastoma cell by activating caspase death pathway. These findings suggested that celastrol might be a potential novel anti-neuroblastoma agent with minor cytotoxicity.展开更多
In the present study, we aimed to investigate the effects of key extraction parameters including extraction time（10–20 min）, extraction temperature（30–60 °C）, ultrasonic power（60–90 W） and solvent-to-so...In the present study, we aimed to investigate the effects of key extraction parameters including extraction time（10–20 min）, extraction temperature（30–60 °C）, ultrasonic power（60–90 W） and solvent-to-solid（S/S） ratio（10–30 m L/g） on yield of celastrol from Celastrus monospermus Roxb. To optimize the conditions, we investigated the effects of parameters on the ultrasound assisted extraction（UAE） with the Box-Behnken Design（BBD）, one widely used form of Response Surface Methodology（RSM）. In all tested solvents, ethanol was the most effective for celastrol extraction, followed by methanol, ethanol, ethyl acetate, n-butanol and water. A second order polynomial model was fitted well to the extraction experimental data with R2 of 0.9928. Extraction yield of 3.116 mg/g was obtained for celastrol under the optimized extraction conditions of extraction time（20 min）, extraction temperature（46 °C）, ultrasonic power（60 W） and S/S ratio（30 m L/g）. Experimental validation was performed, and the experimental values agreed well with the predicted values. The results indicated that the UAE was good extraction material for celastrol from C. monospermus Roxb.展开更多
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 cela...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.展开更多
基金supported by the National Nature Science Foundation Project(32202067)the Young Talent Fund of Association for Science and Technology in Shaanxi,China(20230203)。
文摘Morin is a functional flavonoid commonly found in human diet.Compared to being used solely,it is evident that morin can be more effective as a drug adjuvant.However,research on the combined effect and its corresponding mechanism is limited.Here,we found that morin significantly potentiated the inhibitory effects of the natural compound celastrol on the proliferation of lung cancer cells.Morin and celastrol synergistically exhibit marked apoptosis induction in lung cancer cells,accompanied by changes in the abundance of apoptosis-related proteins.Transcriptome analyses revealed that the combination of morin and celastrol had a significant impact on the number of differentially expressed genes in lung cancer cells.Among these genes,BIRC3 was one of the most significantly different ones,which plays a crucial role in the process of tumor resistance to apoptosis.In addition,several genes identified are primarily associated with intracellular signal transduction pathways,specifically the NF-κB signaling pathway.Importantly,the treatment combining morin and celastrol in tumor-bearing mice results in a synergistic effect that significantly suppressed tumor growth.These findings indicate that morin could be a promising functional adjuvant,and the combination of morin and celastrol has potential for the treating lung cancer.
基金supported by National Natural Science Foundation of China(No.81973662)National Interdisciplinary Innovation Team of Traditional Chinese Medicine(No.ZYYCXTDD-202209)+3 种基金Postdoctoral Fellowship Program of CPSF(No.GZC20230333)Sichuan Science and Technology Program(No.2023NSFSC1195)Central Guidance on Local Science and Technology Development Fund of Sichuan(No.23ZYZYTS0420)Central Nervous System Drug Key Laboratory of Sichuan Province(No.230046-01SZ).
文摘Combining cytotoxic drugs with tumor microenvironment(TME)modulator agents is an effective strategy to enhance anti-tumor effects.In this study,two natural anti-tumor active ingredients celastrol(CEL)and glycyrrhetinic acid(GA)were combined for tumor treatment.In order to ensure the precise co-delivery and controllable synchronous release of combined drugs to tumors,it is necessary to construct a suitable nano-drug delivery platform.Based on this,we coupled hyaluronic acid(HA)with CEL by amide reaction to obtain an amphiphilic polymer prodrug HA-SS-CEL,and GA was spontaneously loaded into polymer micelles by self-assembly to obtain G/HSSC-M.G/HSSC-M has ideal size distribution,redox-responsive synchronous drug release,enhanced tumor cell internalization and in vivo tumor targeting.Compared with free drugs,the construction of multifunctional polymer micelles makes G/HSSC-M show better anticancer effect at the same concentration,and can significantly inhibit the proliferation and migration of HepG2 and 4T1 cells.In the in vivo experiments,G/HSSC-M achieved a tumor inhibition rate as high as 75.12%in H22 tumor-bearing mice.The mechanism included regulation of M1/M2 macrophage polarization,inhibition of Janus kinase 1/signal transducer and activator of transcription 3(JAK1/STAT3)signaling pathway,and remodeling of tumor blood vessels.Therefore,the development of prodrug micelles coloaded with CEL and GA provides a promising drug co-delivery strategy for combined cancer therapy.
基金Hangzhou Municipal Health Commission Project(A20231278)Zhejiang Province Medical and Health Technology Plan Project(2024KY219)+1 种基金Zhejiang Traditional Medicine and Technology Program,China(2024ZR114)Peak Discipline Nephrology of Hangzhou(Integrated Traditional and Western Medicine,2025HZGF12).
文摘Objective:To investigate the mechanisms underlying the renoprotective effects of celastrol,a bioactive compound extracted from the traditional Chinese medicinal plant Tripterygium wilfordii in diabetic kidney disease(DKD).Methods:We established a DKD model using db/db mice and investigated the protective mechanisms of celastrol against DKD progression using integrated analysis of 16S rRNA sequencing and transcriptome analysis.We evaluated colon tissue damage using hematoxylin and eosin and immunofluorescence staining.In addition,16S rRNA sequencing and transcriptomic analyses were performed to explore the potential mechanisms of celastrol.Immunofluorescence staining,Western blotting and real-time quantitative polymerase chain reaction analysis were performed to confirm the PPAR signaling pathway related proteins in kidney tissues.Results:Celastrol alleviated colon injury and increased the expression of mucosal barrier markers,particularly occludin and zonula occludens-1.The 16S rRNA gene sequencing analysis demonstrated that treatment with celastrol altered the diversity and abundance of the gut microbiota.Spearman’s correlation analysis further revealed significant associations among gut microbial,renal injury markers,and serum lipid profiles.A subsequent renal transcriptome analysis revealed that celastrol significantly modulated the renal transcriptional landscape,primarily by regulating genes associated with the PPAR signaling pathway and lipid metabolism.Further investigations demonstrated that celastrol significantly downregulated adipose differentiation-related protein expression and attenuated DKD progression by activating the PPAR pathway.Conclusions:This study demonstrates that celastrol alleviates both colonic and renal injuries by modulating the gut-kidney axis through PPAR-mediated lipid metabolism regulation,indicating its potential as a therapeutic approach for DKD management.
基金Supported by National Natural Science Foundation of China,No.82002609.
文摘BACKGROUND The role of NLR family pyrin domain containing 3(NLRP3)in post-endoscopic submucosal dissection(ESD)esophageal stricture remains incompletely understood.The effect of celastrol(CEL)on the prevention of esophageal strictures has not yet been investigated.AIM To explore the effect of CEL on the prevention of esophageal stricture in rats.METHODS NLRP3,interleukin(IL)-1β,and IL-18 mRNA levels were measured in patients’tissues after esophageal ESD.NLRP3 expression in esophageal fibroblasts was determined using immunohistochemistry and immunofluorescence staining.Lentiviral transfection was used to induce NLRP3 overexpression and thioredoxin reductase 1(TXNRD1)silencing.The CCK8 assay was used to determine the optimal CEL concentration.Reactive oxygen species(ROS)generation was detected via fluorescence and flow cytometry.Masson’s trichrome staining and barium esophagography were performed to assess collagen deposition and esophageal stenosis.RESULTS The mRNA levels of NLRP3 and IL-1βwere higher in human tissues from the ESD resection bed than in normal esophageal mucosa.NLRP3 overexpression in primary rat esophageal fibroblasts led to high collagen 1 expression.Thus,NLRP3 participated in esophageal inflammation and tissue repair after ESD.Comparable to prednisolone,CEL significantly inhibited NLRP3 activation in vitro and in vivo,and esophageal strictures were markedly alleviated.Mechanistically,CEL upregulated TXNRD1 expression and reduced ROS production,thereby inhibiting NLRP3 expression.This effect was reversed by TXNRD1 silencing.Furthermore,TXNRD1 interacted with NLRP3 and promoted its ubiquitination.CONCLUSION CEL is a promising alternative therapeutic agent for the prevention of post-ESD esophageal strictures.
基金the Shanghai Gongli Hospital Youth Project (No. 2014GLQN16 for YS and No. 2012GLQN09 for XZ)the Shanghai Pudong District Science and Technology Innovation Project (No. PKJ2013-Y03 for YW)+5 种基金the Shanghai Pudong Youth Talent Project in Medicine (No. PWRq2013-11 for FC)the Shanghai Yang Fan Project (No. 15YF1410800 for FC)the International Science & Technology Cooperation Project of China (Grant 2011DFB30010 for GU)the National Natural Science Foundation of China (No. 81102349 for BP and No. 81400793 for YW)the Shanghai Excellent Academic Leader in Medicine (No. XBR2011054 for DZ)the Shanghai Traditional Chinese Medicine Content Construction Innovation Project (No. ZY3-CCCX-3-7001 for DZ)
文摘OBJECTIVE： Celastrol has been established as a nuclear factor-κB（NF-κB） activation inhibitor; however, the exact mechanism behind this action is still unknown. Using text-mining technology, the authors predicted that int erleukin-1 receptor-associated kinases（IRA Ks） are potential celastrol targets, and hypothesized that targeting IRAKs might be one way that celastrol inhibits NF-κB. This is because IRAKs are key molecules for some crucial pathways to activate NF-κB（e.g., the inter leukin-1 receptor（IL-1R）/Toll- like receptor（TLR） superfamily）.METHODS： The human hepatocellular cell line（Hep G2） treated with palmitic acid（PA） was used as a model for stimulating TLR4/NF-κB activation, in order to observe the potential effects of celastrol in IRAK regulation and NF-κB inhibition. The transfection of small interfering RNA was used for down-regulating TLR4, IRAK1 and IRAK4, and the Western blot method was used to detect changes in the protein expressions.RESULTS： The results showed that celastrol could effectively inhibit PA-caused TLR4-dependent NF-κB activation in the Hep G 2 cells; PA also activated IRAKs, which were inhibited by celastrol. Knocking down IRAKs abolished PA-caused NF-κB activation.CONCLUSION： The results for the first time show that targeting IRAKs is one way in which celastrol inhibits NF-κB activation.
基金Supported by National Natural Science Foundation of China (No.81300749)Guangdong Province Natural Science Foundation (No.2018A030313628)+1 种基金973 program (No.2015CB964600)the State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University
文摘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.
基金support from the National Natural Science Foundation of China(Grants No.82304827,82074098,81841001)the Fundamental Research Funds for the Central public welfare research institutes(ZZ13-ZD-07),the National Key Research and Development Programof China(2020YFA0908000,2022YFC2303600)+7 种基金the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No:ZYYCXTD-C-202002)The Shenzhen Medical Research Fund of Shenzhen Medical Academy of Research and Translation(B2302051)the Fundamental Research Funds for the Central Public Welfare Research Institutes(Grants No.ZZ13-YQ-108)the Shenzhen Science and Technology Innovation Commission(Grants No.JCYJ20210324115800001)the Science and Technology Foundation of Shenzhen(Shenzhen Clinical Medical Research Center for Geriatric Diseases),the Distinguished Expert Project of Sichuan Province Tianfu Scholar(CW202002)Supported by Shenzhen Governmental Sustainable Development Fund(KCXFZ20201221173612034)Supported by Shenzhen key Laboratory of Kidney Diseases(ZDSYS201504301616234)Supported by Shenzhen Fund for Guangdong Provincial High-level Clinical Key Specialties(NO.SZGSP001).
文摘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.
基金the National Natural Science Foundation of China(Grant No.:81973305)the Science and Technology Planning Project of Guangzhou,China(Grant No.:201904010487)+1 种基金the Natural Science Foundation of Guangdong Province,China(Grant No.:2021A1515010897)the Discipline Construction Fund of Central People’s Hospital of Zhanjiang(Grant Nos.:2020A01 and 2020A02).
文摘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.
基金supported by the Key Research and Development Plan of Jiangsu Province(Social Development Project,No.BE2021679)the National Natural Science Foundation Project(No.82171762).
文摘Excessive oxidative stress impairs cartilage matrix metabolism balance,significantly contributing to osteoarthritis(OA)development.Celastrol(CSL),a drug derived from Tripterygium wilfordii,has recognized applications in the treatment of cancer and immune system disorders,yet its antioxidative stress mechanisms in OA remain underexplored.This study aimed to substantiate CSL’s chondroprotective effects and unravel its underlying mechanisms.We investigated CSL’s impact on chondrocytes under both normal and inflammatory conditions.In vitro,CSL mitigated interleukin(IL)-1β-induced activation of proteinases and promoted cartilage extracellular matrix(ECM)synthesis.In vivo,intra-articular injection of CSL ameliorated cartilage degeneration and mitigated subchondral bone lesions in OA mice.Mechanistically,it was found that inhibiting nuclear factor erythroid 2-related factor 2(NRF2)abrogated CSL-mediated antioxidative functions and exacerbated the progression of OA.This study is the first to elucidate the role of CSL in the treatment of OA through the activation of NRF2,offering a novel therapeutic avenue for arthritis therapy.
基金supported by grants from the National Natural Science Foundation of China(No.21705137)China and donation from Kwok Chung Bo Fun Charitable Fund for the establishment of the Kwok Yat Wai Endowed Chair of Environmental and Biological Analysis。
文摘Inflammation is a defense mechanism associated with a wide range of diseases.Celastrol is a small molecule isolated from traditional Chinese medicine with potent anti-inflammation activity.In this study,we established an integrated quantitative proteomics strategy to investigate the acute response to celastrol treatment in a rat macrophage cell line challenged with lipopolysaccharide(LPS).Both stableisotopic based non-targeted quantitative profiling and PRM-based targeted quantitation methods were employed.Dimethyl-labeling based non-targeted profiling revealed 28 and 52 proteins that significantly up-and down-regulated by celastrol.Bioinformatics analysis pinpoint key signaling pathways affected.Seven proteins were selected for examining their time-dependent regulatory pattern in response to celastrol using targeted PRM quantitation.The abundance of mRNA at multiple time-points of selected proteins was also examined.Celastrol induced an acute response of selected key transcriptional factors in terms of mRNA or protein abundance within one hour.Interestingly,regulatory trend of mRNA and protein abundance suggested a novel dual mechanism of celastrol in the terms of acute antiinflammation.The integrated quantitative proteomic strategy established in this study constitutes an efficient workflow to characterize key components and their time-dependent regulatory pattern for monitoring drug response.
基金This work was supported by the National Natural Science Youth Foundation of China(No.81602204).
文摘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.
基金Supported by Nanjing Medical Science and technique Development Foundation (No.QRX17194)。
文摘OBJECTIVE: To investigate the efficacy of celastrol treatment of hepatocellular carcinoma(HCC) cells in vitro and in vivo and to propose a mechanism of action.METHODS: A human Hep G2 liver cancer cell line and a xenograft tumor model were used to investigate the effects of celastrol on HCC in vitro and in vivo. A CCK-8 kit was used to detect cell viability.Flow cytometry and terminal-deoxynucleoitidyl transferase mediated nick end labeling staining were used to detect apoptosis. Western blotting and immunohistochemistry were used to detect the expression of cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, cleaved-PARP, mammalian target of rapamycin(mTOR), and p-mTOR. Hematoxylin-eosin staining was used to observe the tissue morphology.RESULTS: Celastrol decreased the viability of Hep G2 cells and induced apoptosis. Western blot assays indicated that celastrol up-regulated cleaved-caspase-3, cleaved-caspase-8, cleaved-caspase-9, and cleaved-PARP by inhibiting the phosphorylation of mTOR in Hep G2 cells. Moreover,celastrol inhibited the tumor growth in a xenograft model. Celastrol also induced caspase-dependent apoptosis(up-regulation of cleaved-caspase-3,-8,-9, and cleaved-PARP) and inhibited the activation of mTOR in vivo.CONCLUSION: Celastrol induces caspase-dependent apoptosis in HCC cells by inhibiting the activation of mTOR.
基金supported by the National Natural Science Foundation of China(Nos.81600775,U1904176 and 21504082)the Science and Technology Program of Henan Province,China(No.202102310072)the Medical Science and Technology Program of Henan Province,China(Nos.2018020398 and SB201902026)。
文摘Celastrol,a Chinese herbal medicine,has exhibited anticancer activity in many types of cancer cells.However,the further clinical application of celastrol is restricted by its poor water solubility and serious side effects.Furthermore,the apoptosis mechanism of tumor cells induced by celastrol has not been exhausted yet.In this study,we developed a reduction sensitive polymeric vector for tumor-targeted celastrol delivery.And our researches indicated that the celastrol could be delivered by reductionsensitive nanomedicine(RSNMs)with a controlled release strategy.Meanwhile,the cell uptake results indicated that excellent reduction-sensitive behavior of RSNMs could effectively accelerate celastrol into the human retinoblastoma(RB)cell.The cell cytotoxicity assay demonstrated that celastrol inhibited proliferation of human RB Y79 cells growth in a dose-dependent manner.Furthermore,the results of flow cytometry and terminal dUTP nick-end labeling(TUNEL)staining showed that celastrol induced apoptosis of the RB Y79 cells,and revealed a time-dependent increase in apoptosis induction of RB Y79 cells.The results of western blotting showed that celastrol induced the apoptosis of human RB Y79 cells involving the activation of caspase-3 and caspase-9.In conclusion,our results revealed that RSNMs may be utilized as a novel therapy for retinoblastoma.
基金supported by the National Key R&D Program of China(No.2020YFA0908000)the National Natural Science Foundation of China(Nos.81773830 and 81973418)the Key Project at Central Government Level:the ability establishment of sustainable use for valuable Chinese medicine resources(No.2060302-1806-03),and the National Program for Special Support of Eminent Professionals.
文摘Tripterygium hypoglaucum(Levl.)Hutch,a traditional Chinese medicinal herb with a long history of use,is widely distributed in China.One of its main active components,celastrol,has great potential to be developed into anti-cancer and anti-obesity drugs.Although it exhibits strong pharmacological activities,there is a lack of sustainable sources of celastrol and its derivatives,making it crucial to develop novel sources of these drugs through synthetic biology.The key step in the biosynthesis of celastrol is considered to be the cyclization of 2,3-oxidosqualene into friedelin under the catalysis of 2,3-oxidosqualene cyclases.Friedelin was speculated to be oxidized into celastrol by cytochrome P450 oxidases(CYP450s).Here,we reported a cytochrome P450 ThCYP712K1 from Tripterygium hypoglaucum(Levl.)Hutch that catalyzed the oxidation of friedelin into polpuonic acid when heterologously expressed in yeast.Through substrate supplementation and in vitro enzyme analysis,ThCYP712K1 was further proven to catalyze the oxidation of friedelin at the C-29 position to produce polpunonic acid,which is considered a vital step in the biosynthesis of celastrol,and will lay a foundation for further analysis of its biosynthetic pathway.
基金supported by Shahrekord University of Medical Sciences,Shahrekord,Iran(Ethics Code:IR.SKUMS.REC.1397.119,Grant No.3696 and Ethics Code:IR.SKUMS.REC.1401.197,Grant No.6651).
文摘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.
基金suppor ted by the National Key Research and Development Program of China(2020YFA0908000)the Innovation Team and Talents Cultivation Program of the National Administration of Traditional Chinese Medicine(ZYYCXTD-C-202002)+1 种基金the National Natural Science Foundation of China(82074098,81841001)the Fundamental Research Funds for the Central Public Welfare Research Institutes(ZXKT18003)。
文摘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.
基金Supported by National Natural Science Foundation of China(No.81570826)。
文摘AIM:To evaluate celastrol's effect on choroidal neovascularization(CNV).METHODS:In this study,neovascular formation in vitro(tube formation and aortic ring culture)and in vivo(laser induced neovascular in mice)was treated with celastrol to evaluate this natural compound's impact on CNV.Western blot was applied to explore the possible mechanism for it.For in vitro assay,triplicate for each group was repeated at least three times.For in vivo assay,each group contains 5 mice.RESULTS:Celastrol supressed tube formation and aortic ring sprout neovascularization.In vitro assay exhibited that celastrol inhibiting vascular endothelial growth factor(VEGF)-induced proliferation and migration of human umbilical vein endothelial cells and human choroidal endothelial cells,and by blocking VEGF signaling.Furthermore,intraperitoneal administration of celastrol significantly reduced the area of laser-induced CNV in an in vivo mouse model.By day 14,the area of CNV had decreased by 49.15%and 80.26%in the 0.1 mg/kg celastrol-treated group(n=5)and in the 0.5 mg/kg celastrol treated group(n=5),respectively,compared to the vehicle-treated group(n=5).CONCLUSION:Celastrol inhibits CNV by inhibiting VEGF-induced proliferation and migration of vascular endothelial cells,indicating that celastrol is a potent,natural therapeutic compound for the prevention of CNV.
基金Supported by the Scientific Research Project of Shaanxi Provincial Department of Health:the Protective Role of Celastrol and its Active Mechanisms in Neuroblastoma Treatment (2020A1515003129)。
文摘OBJECTIVE: To investigate the in-depth pharmacological mechanisms of celastrol in children neuroblastoma treatment. METHODS: In the current study, we examined the effects of celastrol on children neuroblastoma cells viability and proliferation by cell counting kit-8 assay and colony formation assay. Annexin V-FTIC and PI staining were applied to determine cell apoptosis after celastrol treatment. ROS generation levels were examined by 2′, 7′-dichloroflfluorescin diacetate. RESULTS: We found that celastrol could suppress the proliferation of children neuroblastoma cells with few effects on normal cell lines in vitro. Further mechanisms studies have shown that celastrol inhibited cell cycle progression and induced cell apoptosis in QDDQ-NM and SH-SY5Y cells. In addition, ROS production might involve in celastrol-mediated apoptotic cell death in children neuroblastoma cells by activating caspase death pathway. CONCLUSIONS: Our findings demonstrated that celastrol could promote ROS generation-induced apoptosis in neuroblastoma cell by activating caspase death pathway. These findings suggested that celastrol might be a potential novel anti-neuroblastoma agent with minor cytotoxicity.
文摘In the present study, we aimed to investigate the effects of key extraction parameters including extraction time（10–20 min）, extraction temperature（30–60 °C）, ultrasonic power（60–90 W） and solvent-to-solid（S/S） ratio（10–30 m L/g） on yield of celastrol from Celastrus monospermus Roxb. To optimize the conditions, we investigated the effects of parameters on the ultrasound assisted extraction（UAE） with the Box-Behnken Design（BBD）, one widely used form of Response Surface Methodology（RSM）. In all tested solvents, ethanol was the most effective for celastrol extraction, followed by methanol, ethanol, ethyl acetate, n-butanol and water. A second order polynomial model was fitted well to the extraction experimental data with R2 of 0.9928. Extraction yield of 3.116 mg/g was obtained for celastrol under the optimized extraction conditions of extraction time（20 min）, extraction temperature（46 °C）, ultrasonic power（60 W） and S/S ratio（30 m L/g）. Experimental validation was performed, and the experimental values agreed well with the predicted values. The results indicated that the UAE was good extraction material for celastrol from C. monospermus Roxb.
基金the National Key Research and Development Program of China(2020YFA0908000,2022YFC2303600)the National Natural Science Foundation of China(81903866,82274182)and the Central Public Welfare Research Institutes(ZZ13-YQ-105,ZZ15-YQ-065,ZZ14-YQ-058).
文摘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.
