Background:Cisplatin(CDDP)is a cornerstone chemotherapeutic agent for many solid tumors,but its clinical use is severely limited by dose-dependent nephrotoxicity,which results in acute kidney injury(AKI)in a significa...Background:Cisplatin(CDDP)is a cornerstone chemotherapeutic agent for many solid tumors,but its clinical use is severely limited by dose-dependent nephrotoxicity,which results in acute kidney injury(AKI)in a significant proportion of patients.CDDP-induced AKI involves interconnected mechanisms,including inflammation,oxidative stress,and tubular cell death.In this study,we aimed to investigate the renoprotective effects of esculetin(ES),a natural antioxidant coumarin,in a murine model of CDDP-induced AKI.Methods:Male C57BL/6 mice(8–10 weeks)received a single intraperitoneal injection of CDDP(20 mg/kg)with or without ES(40 mg/kg/day,oral gavage).Renal function,histopathology,and molecular markers of inflammation,oxidative stress,mitogen-activated protein kinase(MAPK)activation,endoplasmic reticulum(ER)stress,apoptosis,and ferroptosis were assessed by standard biochemical,histological,and immunoblotting techniques.Results:ES significantly reduced CDDP-induced elevations in serum creatinine and blood urea nitrogen,preserved renal structure,and decreased histological injury scores.Molecular analyses showed that ES suppressed the production of systemic and renal proinflammatory cytokines and inhibited the expression of chemokines and adhesion molecules.ES also suppressed the phosphorylation of extracellular signal-regulated kinase 1/2 and p38 MAPKs,mitigating stress-induced inflammatory and apoptotic signaling.Additionally,ES treatment reduced the expression of unfolded protein response markers,such as C/EBP homologous protein,which is indicative of alleviated ER stress.Oxidative injury was reduced,as evidenced by lower malondialdehyde and 4-hydroxynonenal levels and restored glutathione content.Importantly,ES mitigated ferroptosis,as demonstrated by decreased expression of pro-ferroptotic markers and preservation of anti-ferroptotic mediators,including glutathione peroxidase 4 and solute carrier family 7member 1.Conclusion:Collectively,our findings provide the first in vivo evidence that ES robustly protects against CDDP-induced AKI by simultaneously targeting oxidative stress,inflammation,MAPK,and ER stress pathways,apoptosis,and ferroptosis.These results highlight ES as a potential candidate for preventing CDDP-induced nephrotoxicity.展开更多
Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form...Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form of regulated cell death,has garnered considerable attention due to its lethal effect on tumor cells.However,the exact role of ferroptosis in esculetin-mediated anti-hepatocellular carcinoma(HCC)effects remains poorly understood.This study investigated the impact of esculetin on HCC cells both in vitro and in vivo.The findings indicate that esculetin effectively inhibited the growth of HCC cells.Importantly,esculetin promoted the accumulation of intracellular Fe^(2+),leading to an increase in ROS production through the Fenton reaction.This event subsequently induced lipid peroxidation(LPO)and triggered ferroptosis within the HCC cells.The occurrence of ferroptosis was confirmed by the elevation of malondialdehyde(MDA)levels,the depletion of glutathione peroxidase(GSH-Px)activity,and the disruption of mitochondrial morphology.Notably,the inhibitor of ferroptosis,ferrostatin-1(Fer-1),attenuated the anti-tumor effect of esculetin in HCC cells.Furthermore,the findings revealed that esculetin inhibited the Nrf2-xCT/GPx4 axis signaling in HCC cells.Overexpression of Nrf2 upregulated the expression of downstream SLC7A11 and GPX4,consequently alleviating esculetin-induced ferroptosis.In conclusion,this study suggests that esculetin exerts an anti-HCC effect by inhibiting the activity of the Nrf2-xCT/GPx4 axis,thereby triggering ferroptosis in HCC cells.These findings may contribute to the potential clinical use of esculetin as a candidate for HCC treatment.展开更多
Esculetin,a natural derivative from the traditional and widely-used Chinese medicinal herb Cortex Fraxini,has a variety of pharmacological effects,especially in anti-inflammation.However,it is not clear whether escule...Esculetin,a natural derivative from the traditional and widely-used Chinese medicinal herb Cortex Fraxini,has a variety of pharmacological effects,especially in anti-inflammation.However,it is not clear whether esculetin has a therapeutic effect on sepsis.This study aimed to investigate the anti-inflammatory and protective effects of esculetin on early sepsis.The results showed that the lung injury was significantly relieved with the treatment of esculetin,accompanied with the restrained production of inflammatory factors including IL-1β,IL-6,TNF-α,CCL2 and iNOS during the early phase of E.coli-induced sepsis.Of note,activation of NF-κB and STAT1/STAT3 signals,the main upstream signals of many inflammatory factors,were attenuated by esculetin in both lung tissues from septic mice and LPS-stimulated macrophage.These findings suggested that the protection of esculetin against early sepsis should be related to its anti-inflammatory effect,which was at least partly due to its inhibition on NF-κB and STAT1/STAT3 signaling pathway in macrophage.Thus,esculetin could serve as a potential therapeutic agent by rebalancing innate immune response in macrophage for the treatment of early sepsis.展开更多
OBJECTIVE: To determine the effect of an esculetin formulation(at 97.4% purity) on osteoporosis, and to investigate the potential underlying molecular mechanism(s).METHODS: Sixty specific pathogen free-grade female Wi...OBJECTIVE: To determine the effect of an esculetin formulation(at 97.4% purity) on osteoporosis, and to investigate the potential underlying molecular mechanism(s).METHODS: Sixty specific pathogen free-grade female Wistar rats were randomly assigned to three groups: blank control(n = 12), sham(n = 12), and model(n = 36). The model group were bilaterally ovariectomized. The sham group had the tissue surrounding the ovaries removed, while the ovaries were retained. After 3 months, the model group was randomly divided into three subgroups: OVX(n = 12), positive control(n = 12), and esculetin(n =12). The positive control group and the esculetin group were intragastrically administered diethylstilbestrol(0.046 mgkd^(-1)), respec·kg^(-1)tively,·d^(-1)) or esculetin(384 mgg^(-1) once per day for 6 consecu··-tive days; medication administration was then stopped for 1 d, before being administered for another 6 consecutive days. All rats were treated for 3months. Samples were collected at the end of the treatment period. An Osteocore3 Digital 2D bone densitometer was used to test the bone mineral density, and histomorphometric analysis was performed to measure bone mass, bone formation,and bone resorption. Enzyme-linked immunosorbent assay analysis was used to measure the serum concentrations of interleukin-6(IL-6), osteoprotegerin(OPG), and receptor activator of nuclear factor-kappa B ligand(RANKL). Immunohistochemistry and in situ hybridization were performed to detect the protein and mRNA expressions of OPG and RANKL in osteoblasts and bone marrow stromal cells.RESULTS: Compared with the OVX group, the esculetin group had significantly greater femoral bone mineral density and tibial trabecular bone volume,and significantly smaller trabecular resorption surface. The percentage of trabecular formation surface, average osteoid width, trabecular bone mineralization rate, and cortical bone mineralization rate did not significantly differ between groups. Compared with the sham group, the esculetin group had significantly decreased serum levels of IL-6and RANKL, and significant downregulation of RANKL protein and mRNA expression levels in osteoblasts and bone marrow stromal cells; however, there was no significant difference between groups in OPG.CONCLUSION: Esculetin can increase bone mass by upregulating RANKL expression in osteoblasts and bone marrow stromal cells, and decreasing serum IL-6 concentration. This indicates that the therapeutic effect of esculetin on osteoporosis occurs via decreased bone resorption.展开更多
Coumarins belong to a diverse group of naturally occurring non-nutrient phytochemicals known as benzo-α- pyrones. In this study, esculetin, a 6,7-dihydroxy derivative of coumarin with pleiotropic biological activitie...Coumarins belong to a diverse group of naturally occurring non-nutrient phytochemicals known as benzo-α- pyrones. In this study, esculetin, a 6,7-dihydroxy derivative of coumarin with pleiotropic biological activities, was found to have no significant cytotoxic effect on normal murine macrophages, but it could increase the in vivo migration of the thioglycollate-elicited macrophages in a dose-dependent manner. Moreover, esculetin significantly increased the endocytic activity, and augmented the nitric oxide production and iNOS gene expression in LPS-treated macrophages. In addition, in vivo administration of esculetin into mice was shown to increase the mitogenesis of splenic lymphocytes towards Con A and LPS stimulations, and induced the LAK activity of splenic lymphocytes. Collectively, our results indicate that esculetin could exert immunomodulatory effects on murine macrophages and lymphocytes, both in vitro and in vivo, and this might be one of the possible mechanisms by which coumarins can exert their chemopreventive and anti-tumor activities in vivo. Cellular ＆ Molecular Immunology. 2005;2（3）： 181-188.展开更多
Molecularly imprinted polymers(MIPs)for solid-phase extraction and pre-concentration of esculetin have been successfully prepared by the bulk polymerization method using esculetin as a template molecule.Polymers of va...Molecularly imprinted polymers(MIPs)for solid-phase extraction and pre-concentration of esculetin have been successfully prepared by the bulk polymerization method using esculetin as a template molecule.Polymers of varying composition were prepared using different monomers(4-vinylpyridine,methacrylic acid,and acrylamide),ethylene glycol dimethacrylate as the cross-linker,2,2-azobis(2-methylpropinitrile)as the initiator,and different porogen solvents(ethanol,acetone/methanol,and acetonitrile).The best polymer was obtained when 4-vinylpyridine was used as the monomer and acetone/methanol(3:2)as the porogen solvent,whereas the template:-monomer:-cross-linker ratio was 1:4:20.The imprinting factor of the selected MIPs for esculetin was 3.77.The polymers were evaluated according to their selective recognition properties for esculetin and structurally-related compounds(esculin,scopoletin,coumarin,and 7-methoxycoumarin).Chemical and morphological characterizations of the polymers were investigated by FTIR and scanning electron microscope,which confirmed a high degree of polymerization.Surface area,pore volume,and pore size of the polymer were investigated by Brunauer-Emmett-Teller analysis.MIPs were also successfully used as solid-phase adsorbent materials for the extraction of esculetin from tobacco leaves.Esculetin contents in dried tobacco leaves were found to be(9.27±0.17)μg g-1.展开更多
Background Activation of hepatic stellate cells(HSC)leads to initiation and progression of hepatic fibrosis.HSC senescence is inversely correlated with HSC proliferation and activation.Therefore,induction of HSC senes...Background Activation of hepatic stellate cells(HSC)leads to initiation and progression of hepatic fibrosis.HSC senescence is inversely correlated with HSC proliferation and activation.Therefore,induction of HSC senescence may be a strategy to treat hepatic fibrosis.Coumarin-derivatives like esculetin have been suggested to inhibit proliferation of fibrogenic cells.Therefore,in this study we aimed to investigate the effect of esculetin on HSC activation and senescence.Methods Primary rat HSC were used in all experiments.Real-time cell analyzer and BrdU incorporation assay were used to determine HSC proliferation.Gene expression of the senescence-associated genes Cdkn1a(p21),P53,activation markers Acta2,Col1a1 and quiescence markers Pparg and Lrat were measured by RT-qPCR.Senescence associatedβ-Galactosidase(SA-β-Gal)staining was used to identify senescent HSC.Results Our results demonstrate that esculetin increased markers of senescence and decreased proliferation and activation markers in HSC.Protein expression of P21Cip1,accompanied by phosphorylation of Ser473 Akt and Ser9 GSK3βwas increased by esculetin.The effect of esculetin was dependent on PI3K-Akt signaling.Conclusion We conclude that esculetin induces HSC senescence and reverses the activated phenotype of HSC.The induction of senescence depends on PI3K-Akt-GSK3βsignaling.Esculetin could be a potential therapeutic drug for the treatment of hepatic fibrosis by inducing stellate cell senescence.展开更多
基金supported by research grants from Daegu Catholic University in 2024(No.20245001).
文摘Background:Cisplatin(CDDP)is a cornerstone chemotherapeutic agent for many solid tumors,but its clinical use is severely limited by dose-dependent nephrotoxicity,which results in acute kidney injury(AKI)in a significant proportion of patients.CDDP-induced AKI involves interconnected mechanisms,including inflammation,oxidative stress,and tubular cell death.In this study,we aimed to investigate the renoprotective effects of esculetin(ES),a natural antioxidant coumarin,in a murine model of CDDP-induced AKI.Methods:Male C57BL/6 mice(8–10 weeks)received a single intraperitoneal injection of CDDP(20 mg/kg)with or without ES(40 mg/kg/day,oral gavage).Renal function,histopathology,and molecular markers of inflammation,oxidative stress,mitogen-activated protein kinase(MAPK)activation,endoplasmic reticulum(ER)stress,apoptosis,and ferroptosis were assessed by standard biochemical,histological,and immunoblotting techniques.Results:ES significantly reduced CDDP-induced elevations in serum creatinine and blood urea nitrogen,preserved renal structure,and decreased histological injury scores.Molecular analyses showed that ES suppressed the production of systemic and renal proinflammatory cytokines and inhibited the expression of chemokines and adhesion molecules.ES also suppressed the phosphorylation of extracellular signal-regulated kinase 1/2 and p38 MAPKs,mitigating stress-induced inflammatory and apoptotic signaling.Additionally,ES treatment reduced the expression of unfolded protein response markers,such as C/EBP homologous protein,which is indicative of alleviated ER stress.Oxidative injury was reduced,as evidenced by lower malondialdehyde and 4-hydroxynonenal levels and restored glutathione content.Importantly,ES mitigated ferroptosis,as demonstrated by decreased expression of pro-ferroptotic markers and preservation of anti-ferroptotic mediators,including glutathione peroxidase 4 and solute carrier family 7member 1.Conclusion:Collectively,our findings provide the first in vivo evidence that ES robustly protects against CDDP-induced AKI by simultaneously targeting oxidative stress,inflammation,MAPK,and ER stress pathways,apoptosis,and ferroptosis.These results highlight ES as a potential candidate for preventing CDDP-induced nephrotoxicity.
基金supported by the Natural Science Foundations of Fujian Province(Nos.2021J05063 and 2023J01541)a startup grant for High-level Talents of Fujian Medical University(No.XRCZX2021014)。
文摘Esculetin,a natural dihydroxy coumarin derived from the Chinese herbal medicine Cortex Fraxini,has demonstrated significant pharmacological activities,including anticancer properties.Ferroptosis,an iron-dependent form of regulated cell death,has garnered considerable attention due to its lethal effect on tumor cells.However,the exact role of ferroptosis in esculetin-mediated anti-hepatocellular carcinoma(HCC)effects remains poorly understood.This study investigated the impact of esculetin on HCC cells both in vitro and in vivo.The findings indicate that esculetin effectively inhibited the growth of HCC cells.Importantly,esculetin promoted the accumulation of intracellular Fe^(2+),leading to an increase in ROS production through the Fenton reaction.This event subsequently induced lipid peroxidation(LPO)and triggered ferroptosis within the HCC cells.The occurrence of ferroptosis was confirmed by the elevation of malondialdehyde(MDA)levels,the depletion of glutathione peroxidase(GSH-Px)activity,and the disruption of mitochondrial morphology.Notably,the inhibitor of ferroptosis,ferrostatin-1(Fer-1),attenuated the anti-tumor effect of esculetin in HCC cells.Furthermore,the findings revealed that esculetin inhibited the Nrf2-xCT/GPx4 axis signaling in HCC cells.Overexpression of Nrf2 upregulated the expression of downstream SLC7A11 and GPX4,consequently alleviating esculetin-induced ferroptosis.In conclusion,this study suggests that esculetin exerts an anti-HCC effect by inhibiting the activity of the Nrf2-xCT/GPx4 axis,thereby triggering ferroptosis in HCC cells.These findings may contribute to the potential clinical use of esculetin as a candidate for HCC treatment.
基金supported by the Key Project of Scientific Research Fund of Hunan Provincial Education Department(15A177)the Natural Science Foundation of Hunan Province(2019JJ40275).
文摘Esculetin,a natural derivative from the traditional and widely-used Chinese medicinal herb Cortex Fraxini,has a variety of pharmacological effects,especially in anti-inflammation.However,it is not clear whether esculetin has a therapeutic effect on sepsis.This study aimed to investigate the anti-inflammatory and protective effects of esculetin on early sepsis.The results showed that the lung injury was significantly relieved with the treatment of esculetin,accompanied with the restrained production of inflammatory factors including IL-1β,IL-6,TNF-α,CCL2 and iNOS during the early phase of E.coli-induced sepsis.Of note,activation of NF-κB and STAT1/STAT3 signals,the main upstream signals of many inflammatory factors,were attenuated by esculetin in both lung tissues from septic mice and LPS-stimulated macrophage.These findings suggested that the protection of esculetin against early sepsis should be related to its anti-inflammatory effect,which was at least partly due to its inhibition on NF-κB and STAT1/STAT3 signaling pathway in macrophage.Thus,esculetin could serve as a potential therapeutic agent by rebalancing innate immune response in macrophage for the treatment of early sepsis.
基金Supported by Independent Subject Selection Project of the China Academy of Chinese Medical Sciences:Research on the New Function of Cortex Fraxini in the Treatment of Osteoporosis(No.YZ-1411)
文摘OBJECTIVE: To determine the effect of an esculetin formulation(at 97.4% purity) on osteoporosis, and to investigate the potential underlying molecular mechanism(s).METHODS: Sixty specific pathogen free-grade female Wistar rats were randomly assigned to three groups: blank control(n = 12), sham(n = 12), and model(n = 36). The model group were bilaterally ovariectomized. The sham group had the tissue surrounding the ovaries removed, while the ovaries were retained. After 3 months, the model group was randomly divided into three subgroups: OVX(n = 12), positive control(n = 12), and esculetin(n =12). The positive control group and the esculetin group were intragastrically administered diethylstilbestrol(0.046 mgkd^(-1)), respec·kg^(-1)tively,·d^(-1)) or esculetin(384 mgg^(-1) once per day for 6 consecu··-tive days; medication administration was then stopped for 1 d, before being administered for another 6 consecutive days. All rats were treated for 3months. Samples were collected at the end of the treatment period. An Osteocore3 Digital 2D bone densitometer was used to test the bone mineral density, and histomorphometric analysis was performed to measure bone mass, bone formation,and bone resorption. Enzyme-linked immunosorbent assay analysis was used to measure the serum concentrations of interleukin-6(IL-6), osteoprotegerin(OPG), and receptor activator of nuclear factor-kappa B ligand(RANKL). Immunohistochemistry and in situ hybridization were performed to detect the protein and mRNA expressions of OPG and RANKL in osteoblasts and bone marrow stromal cells.RESULTS: Compared with the OVX group, the esculetin group had significantly greater femoral bone mineral density and tibial trabecular bone volume,and significantly smaller trabecular resorption surface. The percentage of trabecular formation surface, average osteoid width, trabecular bone mineralization rate, and cortical bone mineralization rate did not significantly differ between groups. Compared with the sham group, the esculetin group had significantly decreased serum levels of IL-6and RANKL, and significant downregulation of RANKL protein and mRNA expression levels in osteoblasts and bone marrow stromal cells; however, there was no significant difference between groups in OPG.CONCLUSION: Esculetin can increase bone mass by upregulating RANKL expression in osteoblasts and bone marrow stromal cells, and decreasing serum IL-6 concentration. This indicates that the therapeutic effect of esculetin on osteoporosis occurs via decreased bone resorption.
文摘Coumarins belong to a diverse group of naturally occurring non-nutrient phytochemicals known as benzo-α- pyrones. In this study, esculetin, a 6,7-dihydroxy derivative of coumarin with pleiotropic biological activities, was found to have no significant cytotoxic effect on normal murine macrophages, but it could increase the in vivo migration of the thioglycollate-elicited macrophages in a dose-dependent manner. Moreover, esculetin significantly increased the endocytic activity, and augmented the nitric oxide production and iNOS gene expression in LPS-treated macrophages. In addition, in vivo administration of esculetin into mice was shown to increase the mitogenesis of splenic lymphocytes towards Con A and LPS stimulations, and induced the LAK activity of splenic lymphocytes. Collectively, our results indicate that esculetin could exert immunomodulatory effects on murine macrophages and lymphocytes, both in vitro and in vivo, and this might be one of the possible mechanisms by which coumarins can exert their chemopreventive and anti-tumor activities in vivo. Cellular ＆ Molecular Immunology. 2005;2（3）： 181-188.
基金supported by the Yunnan Green Tobacco Production Research(2011YN01)the China Equipment and Education Resources System(CERS-1-75)
文摘Molecularly imprinted polymers(MIPs)for solid-phase extraction and pre-concentration of esculetin have been successfully prepared by the bulk polymerization method using esculetin as a template molecule.Polymers of varying composition were prepared using different monomers(4-vinylpyridine,methacrylic acid,and acrylamide),ethylene glycol dimethacrylate as the cross-linker,2,2-azobis(2-methylpropinitrile)as the initiator,and different porogen solvents(ethanol,acetone/methanol,and acetonitrile).The best polymer was obtained when 4-vinylpyridine was used as the monomer and acetone/methanol(3:2)as the porogen solvent,whereas the template:-monomer:-cross-linker ratio was 1:4:20.The imprinting factor of the selected MIPs for esculetin was 3.77.The polymers were evaluated according to their selective recognition properties for esculetin and structurally-related compounds(esculin,scopoletin,coumarin,and 7-methoxycoumarin).Chemical and morphological characterizations of the polymers were investigated by FTIR and scanning electron microscope,which confirmed a high degree of polymerization.Surface area,pore volume,and pore size of the polymer were investigated by Brunauer-Emmett-Teller analysis.MIPs were also successfully used as solid-phase adsorbent materials for the extraction of esculetin from tobacco leaves.Esculetin contents in dried tobacco leaves were found to be(9.27±0.17)μg g-1.
基金Supported by a grant from the Chinese Scholarship CouncilSupported by a grant from the Mongolian State Training Foundation。
文摘Background Activation of hepatic stellate cells(HSC)leads to initiation and progression of hepatic fibrosis.HSC senescence is inversely correlated with HSC proliferation and activation.Therefore,induction of HSC senescence may be a strategy to treat hepatic fibrosis.Coumarin-derivatives like esculetin have been suggested to inhibit proliferation of fibrogenic cells.Therefore,in this study we aimed to investigate the effect of esculetin on HSC activation and senescence.Methods Primary rat HSC were used in all experiments.Real-time cell analyzer and BrdU incorporation assay were used to determine HSC proliferation.Gene expression of the senescence-associated genes Cdkn1a(p21),P53,activation markers Acta2,Col1a1 and quiescence markers Pparg and Lrat were measured by RT-qPCR.Senescence associatedβ-Galactosidase(SA-β-Gal)staining was used to identify senescent HSC.Results Our results demonstrate that esculetin increased markers of senescence and decreased proliferation and activation markers in HSC.Protein expression of P21Cip1,accompanied by phosphorylation of Ser473 Akt and Ser9 GSK3βwas increased by esculetin.The effect of esculetin was dependent on PI3K-Akt signaling.Conclusion We conclude that esculetin induces HSC senescence and reverses the activated phenotype of HSC.The induction of senescence depends on PI3K-Akt-GSK3βsignaling.Esculetin could be a potential therapeutic drug for the treatment of hepatic fibrosis by inducing stellate cell senescence.
